Chinese Journal of Experimental Traditional Medical Formulae
Volume 30,2024 Issue 24
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2024,30(24):1-9, DOI: 10.13422/j.cnki.syfjx.20241502
Abstract:
Objective This study aims to investigate the effect of Huanglian Jiedutang on brain iron metabolism disorders， myelin damage， and aggressive behavior in vascular dementia （VaD） mice by regulating the F-box leucine-rich repeat protein 5 （FBXL5）/iron regulatory protein 2 （IRP2） pathway.Method Sixty C57BL/6J mice were randomly divided into six groups： sham operation group， model group， risperidone group （2 mg·kg-1·d-1）， and low-dose， medium-dose， and high-dose groups of Huanglian Jiedutang （0.25， 0.5， 1 g·kg-1·d-1）， with 10 mice in each group. The VaD model was established by bilateral carotid artery stenosis （BCAS）. Drug intervention was administered for two weeks starting from the seventh week. Behavioral assessments， including the touch escape and resident-intruder tests， were conducted in the ninth week. After the behavioral tests， ventromedial hypothalamus ventrolateral （VMHvl） tissue samples were collected. Western blot was used to detect the expression of myelin-associated glycoprotein （MAG）， myelin oligodendrocyte glycoprotein （MOG）， myelin basic protein （MBP）， 4-hydroxynonenal （4-HNE）， glutathione peroxidase 4 （GPX4）， transferrin receptor 1 （TFR1）， ferritin light chain （Ft-L）， ferroportin 1 （FPN1）， FBXL5， and IRP2. Immunofluorescence was used to measure MBP fluorescence intensity. Transmission electron microscopy was employed to observe ultrastructural changes in the myelin sheath. Perl's staining was used to detect tissue iron deposition. Malondialdehyde （MDA） and superoxide dismutase （SOD） levels were measured by enzyme linked immunosorbent assay （ELISA）.Result Compared to the sham operation group， the model group exhibited a significant increase in biting， aggression， and irritability scores， along with a reduced latency of the attack （P<0.01）. The expression levels of MAG， MOG， and MBP， as well as the fluorescent density of MBP， were significantly decreased （P<0.01）. Disordered myelin ultrastructure， increased Ft-L and TFR1 expression， decreased FPN1 expression， and elevated iron deposition were observed （P<0.01）. Antioxidants GPX4 and SOD were reduced， while 4-HNE and lipid peroxidation product MDA were increased （P<0.01）. FBXL5 protein expression decreased， and IRP2 protein expression increased （P<0.01）. Compared with the model group， in the middle-dose and high-dose groups of Huanglian Jiedutang and the risperidone group， the number of bites， aggressive behaviors， and irritability scores were reduced （P<0.05， P<0.01）， while the latency of the attack increased （P<0.01）. In the middle-dose and high-dose groups of Huanglian Jiedutang， the expression levels of MAG， MOG， and MBP， as well as MBP immunofluorescence increased （P<0.01）. The ultrastructure was orderly arranged. Ft-L and TFR1 expression decreased； FPN1 expression increased， and iron deposition was reduced （P<0.01）. GPX4 and SOD expression levels increased， and 4-HNE and MDA expression levels decreased （P<0.01）. FBXL5 protein expression increased， and IRP2 protein expression decreased （P<0.01）.Conclusion Huanglian Jiedutang may alleviate brain iron metabolism disorders， myelin damage， and aggressive behavior in VaD mice by regulating FBXL5/IRP2 expression in the VMHvl region.
Objective This study aims to investigate the effect of Huanglian Jiedutang on brain iron metabolism disorders， myelin damage， and aggressive behavior in vascular dementia （VaD） mice by regulating the F-box leucine-rich repeat protein 5 （FBXL5）/iron regulatory protein 2 （IRP2） pathway.Method Sixty C57BL/6J mice were randomly divided into six groups： sham operation group， model group， risperidone group （2 mg·kg-1·d-1）， and low-dose， medium-dose， and high-dose groups of Huanglian Jiedutang （0.25， 0.5， 1 g·kg-1·d-1）， with 10 mice in each group. The VaD model was established by bilateral carotid artery stenosis （BCAS）. Drug intervention was administered for two weeks starting from the seventh week. Behavioral assessments， including the touch escape and resident-intruder tests， were conducted in the ninth week. After the behavioral tests， ventromedial hypothalamus ventrolateral （VMHvl） tissue samples were collected. Western blot was used to detect the expression of myelin-associated glycoprotein （MAG）， myelin oligodendrocyte glycoprotein （MOG）， myelin basic protein （MBP）， 4-hydroxynonenal （4-HNE）， glutathione peroxidase 4 （GPX4）， transferrin receptor 1 （TFR1）， ferritin light chain （Ft-L）， ferroportin 1 （FPN1）， FBXL5， and IRP2. Immunofluorescence was used to measure MBP fluorescence intensity. Transmission electron microscopy was employed to observe ultrastructural changes in the myelin sheath. Perl's staining was used to detect tissue iron deposition. Malondialdehyde （MDA） and superoxide dismutase （SOD） levels were measured by enzyme linked immunosorbent assay （ELISA）.Result Compared to the sham operation group， the model group exhibited a significant increase in biting， aggression， and irritability scores， along with a reduced latency of the attack （P<0.01）. The expression levels of MAG， MOG， and MBP， as well as the fluorescent density of MBP， were significantly decreased （P<0.01）. Disordered myelin ultrastructure， increased Ft-L and TFR1 expression， decreased FPN1 expression， and elevated iron deposition were observed （P<0.01）. Antioxidants GPX4 and SOD were reduced， while 4-HNE and lipid peroxidation product MDA were increased （P<0.01）. FBXL5 protein expression decreased， and IRP2 protein expression increased （P<0.01）. Compared with the model group， in the middle-dose and high-dose groups of Huanglian Jiedutang and the risperidone group， the number of bites， aggressive behaviors， and irritability scores were reduced （P<0.05， P<0.01）， while the latency of the attack increased （P<0.01）. In the middle-dose and high-dose groups of Huanglian Jiedutang， the expression levels of MAG， MOG， and MBP， as well as MBP immunofluorescence increased （P<0.01）. The ultrastructure was orderly arranged. Ft-L and TFR1 expression decreased； FPN1 expression increased， and iron deposition was reduced （P<0.01）. GPX4 and SOD expression levels increased， and 4-HNE and MDA expression levels decreased （P<0.01）. FBXL5 protein expression increased， and IRP2 protein expression decreased （P<0.01）.Conclusion Huanglian Jiedutang may alleviate brain iron metabolism disorders， myelin damage， and aggressive behavior in VaD mice by regulating FBXL5/IRP2 expression in the VMHvl region.
WANG Xiao, WANG Xiaosu, ZHOU Bingduo, XIONG Guangsu, YU Qi, SUN Ji, ZHOU Yun, JING Yi, ZHU Shengliang, LI Li
2024,30(24):10-17, DOI: 10.13422/j.cnki.syfjx.20241122
Abstract:
Objective To observe the clinical efficacy and safety of Chaihu Shugansan combined with Xuanfu Daizhetang （CHSG-XFDZ） in the management of Barrett's esophagus （BE） with liver-stomach disharmony.Method A randomized， parallel， controlled， double-blind clinical trial was conducted. BE patients who met the inclusion criteria were randomized into an observation group and a control group， with 34 patients in each group. The observation group was treated with CHSG-XFDZ combined with omeprazole capsules， and the control group was treated with CHSG-XFDZ mimetic combined with omeprazole capsules. Both groups were treated for 12 weeks. The traditional Chinese medicine （TCM） symptom scores， response rate， BE lesion area， BE pathological changes， and bile acid profile were taken as the indicators to jointly evaluate the clinical efficacy and safety of the two groups.Result A total of 62 patients who completed the trial were included for statistical analysis， including 32 in the observation group and 30 in the control group. There were no statistically significant differences in baseline demographics or disease characteristics between two groups， which suggested that the two groups were comparable. The total response rate in the observation group was 93.7% （30/32）， which was higher than that （60.0%， 18/30） in the control group （χ2=24.766， P<0.05）. After treatment， the response rate regarding the pathological changes in the observation group was 62.5% （20/32）， which was higher than that （23.3%， 7/30） in the control group （χ2=10.270， P<0.05）. The response rate regarding the BE lesion area change in the observation group was 21.9% （7/32）， which had no statistically significant difference from that （6.7%， 2/30） in the control group， which indicated that the advantages of the two regimens were not obvious in terms of reducing the area of BE lesions. Compared with the control group after treatment， the observation group regulated the bile acid profile， which pointed out the direction for further exploring the mechanism of CHSG-XFDZ in treating BE. Neither group showcased adverse reactions with clinical significance during the treatment period.Conclusion CHSG-XFDZ outperformed the control group in terms of alleviating TCM symptoms， ameliorating pathological changes， and improving the bile acid profile in the BE patients with liver-stomach disharmony. It demonstrates certain potential in reducing the lesion area. This formula is safe and effective in treating BE patients with liver-stomach disharmony and deserves further clinical research and widespread application.
Objective To observe the clinical efficacy and safety of Chaihu Shugansan combined with Xuanfu Daizhetang （CHSG-XFDZ） in the management of Barrett's esophagus （BE） with liver-stomach disharmony.Method A randomized， parallel， controlled， double-blind clinical trial was conducted. BE patients who met the inclusion criteria were randomized into an observation group and a control group， with 34 patients in each group. The observation group was treated with CHSG-XFDZ combined with omeprazole capsules， and the control group was treated with CHSG-XFDZ mimetic combined with omeprazole capsules. Both groups were treated for 12 weeks. The traditional Chinese medicine （TCM） symptom scores， response rate， BE lesion area， BE pathological changes， and bile acid profile were taken as the indicators to jointly evaluate the clinical efficacy and safety of the two groups.Result A total of 62 patients who completed the trial were included for statistical analysis， including 32 in the observation group and 30 in the control group. There were no statistically significant differences in baseline demographics or disease characteristics between two groups， which suggested that the two groups were comparable. The total response rate in the observation group was 93.7% （30/32）， which was higher than that （60.0%， 18/30） in the control group （χ2=24.766， P<0.05）. After treatment， the response rate regarding the pathological changes in the observation group was 62.5% （20/32）， which was higher than that （23.3%， 7/30） in the control group （χ2=10.270， P<0.05）. The response rate regarding the BE lesion area change in the observation group was 21.9% （7/32）， which had no statistically significant difference from that （6.7%， 2/30） in the control group， which indicated that the advantages of the two regimens were not obvious in terms of reducing the area of BE lesions. Compared with the control group after treatment， the observation group regulated the bile acid profile， which pointed out the direction for further exploring the mechanism of CHSG-XFDZ in treating BE. Neither group showcased adverse reactions with clinical significance during the treatment period.Conclusion CHSG-XFDZ outperformed the control group in terms of alleviating TCM symptoms， ameliorating pathological changes， and improving the bile acid profile in the BE patients with liver-stomach disharmony. It demonstrates certain potential in reducing the lesion area. This formula is safe and effective in treating BE patients with liver-stomach disharmony and deserves further clinical research and widespread application.
2024,30(24):18-27, DOI: 10.13422/j.cnki.syfjx.20250567
Abstract:
Objective To investigate the pharmacodynamics， pharmacokinetics and brain distribution of seven main components of Jiaotaiwan alone and Jiaotaiwan combined with borneol in lipopolysaccharide（LPS）-induced depression rats.Method Rats were randomly divided into the control group， model group， fluoxetine group（10 mg·kg-1）， Jiaotaiwan group（1.5 g·kg-1） and combination group（1.5 g·kg-1 of Jiaotaiwan+0.05 g·kg-1 of borneol）， with 8 rats in each group. Except for the control group， the depression model was established by intraperitoneal injection of LPS for 7 consecutive days， and each group was given the corresponding drugs or the same volume of pure water by gavage for 14 consecutive days. The behavioral indicators and levels of serum inflammatory factors［interleukin-1β（IL-1β）， IL-6 and tumor necrosis factor-α（TNF-α）］ of rats in each group were compared. The morphological changes of hippocampal neurons were observed by hematoxylin-eosin（HE） and Nissl staining. After the behavioral tests of sucrose preference， open field and forced swimming were completed， blood samples were collected from Jiaotaiwan group and combination group at the set time points after gavage， the contents of seven components in plasma were determined by ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry（UPLC-QqQ-MS/MS）， and the pharmacokinetic parameters of each component were analyzed by DAS 3.2.2. Brains were rapidly removed on ice after blood collection was completed， and UPLC-QqQ-MS/MS was used to compare the content changes of the 7 components in brain tissue.Result Compared with the control group， rats in the model group showed decreased sucrose preference rate and total distance of open field， prolonged swimming immobility time， and increased expression of inflammatory factors in serum（P<0.01）. Compared with the model group， the sucrose preference rate and total distance of open field were increased， and the swimming immobility time was shortened in the rats of each administration group， and the expression of inflammatory factors in serum was decreased in rats from Jiaotaiwan group and combination group（P<0.05， P<0.01）. The results of HE and Nissl staining showed that Jiaotaiwan group and combination group had a certain protective effect on hippocampal neurons. The pharmacokinetic results showed that compared with Jiaotaiwan group， the area under the curve（AUC0-t， AUC0-∞）， peak concentration（Cmax） and the average steady-state plasma concentration（Cav） of berberine and epiberberine in the combination group were increased（P<0.05， P<0.01）， AUC0-t， AUC0-∞， mean residence time（MRT0-∞） and Cav of coptisine were significantly increased（P<0.05， P<0.01）， Cmax of jatrorrhizine increased significantly（P<0.05）， but the pharmacokinetic changes of the seven alkaloids were consistent. The results of brain tissue distribution showed that compared with Jiaotaiwan group， the contents of jatrorrhizine， epiberberine， coptisine， palmatine and berberine in the brain tissue of combination group were increased（P<0.05， P<0.01）， the content of magnoflorine increased and that of berberrubine decreased， but the difference was not statistically significant.Conclusion Jiaotaiwan alone or combined with borneol can improve the depression-like behavior of rats， reduce the levels of serum inflammatory factors， improve the pathological damage of hippocampus， and have antidepressant effect. Compared with Jiaotaiwan alone， the combination of Jiaotaiwan and borneol can increase the exposure of multiple active components of Jiaotaiwan in the plasma and brain tissue of rats， improve its bioavailability， promote its absorption， and improve the brain targeting of the drug， which is more conducive to the antidepressant effect of Jiaotaiwan.
Objective To investigate the pharmacodynamics， pharmacokinetics and brain distribution of seven main components of Jiaotaiwan alone and Jiaotaiwan combined with borneol in lipopolysaccharide（LPS）-induced depression rats.Method Rats were randomly divided into the control group， model group， fluoxetine group（10 mg·kg-1）， Jiaotaiwan group（1.5 g·kg-1） and combination group（1.5 g·kg-1 of Jiaotaiwan+0.05 g·kg-1 of borneol）， with 8 rats in each group. Except for the control group， the depression model was established by intraperitoneal injection of LPS for 7 consecutive days， and each group was given the corresponding drugs or the same volume of pure water by gavage for 14 consecutive days. The behavioral indicators and levels of serum inflammatory factors［interleukin-1β（IL-1β）， IL-6 and tumor necrosis factor-α（TNF-α）］ of rats in each group were compared. The morphological changes of hippocampal neurons were observed by hematoxylin-eosin（HE） and Nissl staining. After the behavioral tests of sucrose preference， open field and forced swimming were completed， blood samples were collected from Jiaotaiwan group and combination group at the set time points after gavage， the contents of seven components in plasma were determined by ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry（UPLC-QqQ-MS/MS）， and the pharmacokinetic parameters of each component were analyzed by DAS 3.2.2. Brains were rapidly removed on ice after blood collection was completed， and UPLC-QqQ-MS/MS was used to compare the content changes of the 7 components in brain tissue.Result Compared with the control group， rats in the model group showed decreased sucrose preference rate and total distance of open field， prolonged swimming immobility time， and increased expression of inflammatory factors in serum（P<0.01）. Compared with the model group， the sucrose preference rate and total distance of open field were increased， and the swimming immobility time was shortened in the rats of each administration group， and the expression of inflammatory factors in serum was decreased in rats from Jiaotaiwan group and combination group（P<0.05， P<0.01）. The results of HE and Nissl staining showed that Jiaotaiwan group and combination group had a certain protective effect on hippocampal neurons. The pharmacokinetic results showed that compared with Jiaotaiwan group， the area under the curve（AUC0-t， AUC0-∞）， peak concentration（Cmax） and the average steady-state plasma concentration（Cav） of berberine and epiberberine in the combination group were increased（P<0.05， P<0.01）， AUC0-t， AUC0-∞， mean residence time（MRT0-∞） and Cav of coptisine were significantly increased（P<0.05， P<0.01）， Cmax of jatrorrhizine increased significantly（P<0.05）， but the pharmacokinetic changes of the seven alkaloids were consistent. The results of brain tissue distribution showed that compared with Jiaotaiwan group， the contents of jatrorrhizine， epiberberine， coptisine， palmatine and berberine in the brain tissue of combination group were increased（P<0.05， P<0.01）， the content of magnoflorine increased and that of berberrubine decreased， but the difference was not statistically significant.Conclusion Jiaotaiwan alone or combined with borneol can improve the depression-like behavior of rats， reduce the levels of serum inflammatory factors， improve the pathological damage of hippocampus， and have antidepressant effect. Compared with Jiaotaiwan alone， the combination of Jiaotaiwan and borneol can increase the exposure of multiple active components of Jiaotaiwan in the plasma and brain tissue of rats， improve its bioavailability， promote its absorption， and improve the brain targeting of the drug， which is more conducive to the antidepressant effect of Jiaotaiwan.
2024,30(24):28-35, DOI: 10.13422/j.cnki.syfjx.20241408
Abstract:
Objective To investigate the therapeutic effect of Qinggongtang in regulating Glu/GABA metabolic balance and the mechanism of its anxiolytic effect on rat models of anxiety.Method Fifty-four rats were randomly divided into normal， model， diazepam （0.225 mg·kg-1）， and low-dose， medium-dose， and high-dose groups of Qinggongtang （5.085， 10.17， 20.34 g·kg-1）， with nine rats in each group. Except for the normal group， the other groups were subjected to indeterminate vacutainer stress and chronic restraint stress for 12 days to prepare the anxiety model. On the 3rd day of the stress， 10 days of corresponding drug intervention was started. At the end of the drug treatment， the anxiety level of rats in each group was evaluated by the elevated cross maze experiment （EPM） and the light and dark box experiment （LDB）， and the effect of Qinggongtang on the anxiety behavior of rats was preliminarily analyzed. The levels of Glu and GABA in the amygdala tissue of the rats were detected by enzyme linked immunosorbent assay （ELISA）， and the changes in the synaptic ultrastructure of the amygdala of the rats in each group were observed by electron microscopy. The mRNA expression of glutamic acid decarboxylase （GAD65 and GAD67）， glutamine synthetase （GS）， and glutamate transporter-1 （GLT-1） in the amygdala were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and their protein expression was detected by Western blot.Result Compared with those in the normal group， rats in the model group showed an obvious anxiety state and dull yellow and lusterless fur. They were irritable， easy to anger， and preferred to curl up in the corner. The number of times the EPM entered the open arm and the residence time in the open arm were significantly reduced （P<0.01）， and the residence time in the open box and the number of times the LDB went through the box were significantly reduced （P<0.01）. The content of Glu in the amygdala was increased （P<0.01）， and the content of GABA was reduced （P<0.01）. The value of Glu/GAB was elevated （P<0.01）， and the number of synaptic and pre-synaptic membrane vesicles in the amygdala was decreased. Sparse dense material in the post-synaptic membrane， increased synaptic gap， slightly disrupted internal structure， and decreased mRNA and protein expressions of GAD65， GAD67， GS， and GLT-1 in the amygdala were observed （P<0.01）. Compared with those in the model group， rats in the medium-dose and high-dose groups of Qinggongtang and the diazepam group had bright fur， sensitive reactions， and more active behavior. The number of times EPM entered the open arm and the residence time in the open arm increased significantly （P<0.01）， and the residence time in the open box and the number of times the LDB went through the box increased significantly （P<0.01）. The content of Glu in all-dose groups of Qinggongtang and the diazepam group decreased （P<0.05， P<0.01）， while GABA content increased （P<0.05， P<0.01）. The value of Glu/GABA decreased （P<0.01）， and the internal and external synaptic structure of each groups of Qinggongtang and the diazepam group was more complete. Synapses and vesicles were numerous， and the synaptic gap was more clearly defined. The efficacy of the high-dose group of Qinggongtang and the diazepam group was the best， and the mRNA and protein expressions of GAD65， GAD67， GS， and GLT-1 in the amygdala were increased in the high-dose group of Qinggongtang and diazepam group （P<0.05， P<0.01）.Conclusion Qinggongtang can improve synaptic plasticity and affect the expression of GAD65， GAD67， GS， and GLT-1 in the amygdala of rats to regulate Glu/GABA metabolic balance and thus exert anxiolytic effects.
Objective To investigate the therapeutic effect of Qinggongtang in regulating Glu/GABA metabolic balance and the mechanism of its anxiolytic effect on rat models of anxiety.Method Fifty-four rats were randomly divided into normal， model， diazepam （0.225 mg·kg-1）， and low-dose， medium-dose， and high-dose groups of Qinggongtang （5.085， 10.17， 20.34 g·kg-1）， with nine rats in each group. Except for the normal group， the other groups were subjected to indeterminate vacutainer stress and chronic restraint stress for 12 days to prepare the anxiety model. On the 3rd day of the stress， 10 days of corresponding drug intervention was started. At the end of the drug treatment， the anxiety level of rats in each group was evaluated by the elevated cross maze experiment （EPM） and the light and dark box experiment （LDB）， and the effect of Qinggongtang on the anxiety behavior of rats was preliminarily analyzed. The levels of Glu and GABA in the amygdala tissue of the rats were detected by enzyme linked immunosorbent assay （ELISA）， and the changes in the synaptic ultrastructure of the amygdala of the rats in each group were observed by electron microscopy. The mRNA expression of glutamic acid decarboxylase （GAD65 and GAD67）， glutamine synthetase （GS）， and glutamate transporter-1 （GLT-1） in the amygdala were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and their protein expression was detected by Western blot.Result Compared with those in the normal group， rats in the model group showed an obvious anxiety state and dull yellow and lusterless fur. They were irritable， easy to anger， and preferred to curl up in the corner. The number of times the EPM entered the open arm and the residence time in the open arm were significantly reduced （P<0.01）， and the residence time in the open box and the number of times the LDB went through the box were significantly reduced （P<0.01）. The content of Glu in the amygdala was increased （P<0.01）， and the content of GABA was reduced （P<0.01）. The value of Glu/GAB was elevated （P<0.01）， and the number of synaptic and pre-synaptic membrane vesicles in the amygdala was decreased. Sparse dense material in the post-synaptic membrane， increased synaptic gap， slightly disrupted internal structure， and decreased mRNA and protein expressions of GAD65， GAD67， GS， and GLT-1 in the amygdala were observed （P<0.01）. Compared with those in the model group， rats in the medium-dose and high-dose groups of Qinggongtang and the diazepam group had bright fur， sensitive reactions， and more active behavior. The number of times EPM entered the open arm and the residence time in the open arm increased significantly （P<0.01）， and the residence time in the open box and the number of times the LDB went through the box increased significantly （P<0.01）. The content of Glu in all-dose groups of Qinggongtang and the diazepam group decreased （P<0.05， P<0.01）， while GABA content increased （P<0.05， P<0.01）. The value of Glu/GABA decreased （P<0.01）， and the internal and external synaptic structure of each groups of Qinggongtang and the diazepam group was more complete. Synapses and vesicles were numerous， and the synaptic gap was more clearly defined. The efficacy of the high-dose group of Qinggongtang and the diazepam group was the best， and the mRNA and protein expressions of GAD65， GAD67， GS， and GLT-1 in the amygdala were increased in the high-dose group of Qinggongtang and diazepam group （P<0.05， P<0.01）.Conclusion Qinggongtang can improve synaptic plasticity and affect the expression of GAD65， GAD67， GS， and GLT-1 in the amygdala of rats to regulate Glu/GABA metabolic balance and thus exert anxiolytic effects.
2024,30(24):36-46, DOI: 10.13422/j.cnki.syfjx.20241743
Abstract:
Objective To clarify the pharmacodynamic characteristics and neuroinflammatory mechanisms of Ruyi Zhenbaowan （RYZBW） in treating nociceptive hypersensitivity and central sensitisation of spinal cord in the mouse model of central post-stroke pain （CPSP）.Method SPF-grade male ICR mice of 8 weeks old were assigned into the sham operation （Sham）， model （CPSP）， low-， medium-， and high-dose （0.303， 0.607 1.214 g·kg-1） RYZBW （RYZBW-L， RYZBW-M， and RYZBW-H， respectively）， and pregabalin （PGB， 0.046 g·kg-1， positive control） groups. The rat model of CPSP was established by injection of type Ⅳ collagenase into the ventral posterior lateral nucleus of the thalamus on day 1. Rats were administrated with corresponding drugs or normal saline （Sham and CPSP groups） by gavage from day 14 to day 17. The mechanical pain sensitivity test was performed on days 0， 3， 4， 7， 10， 14， 17. On day 18， the L5 segment of spinal cord was collected for the detection of inflammatory cytokines by immunoinflammatory microarray， CXC chemokine ligand 16 （CXCL16） by enzyme-linked immunosorbent assay， and calcitonin gene-related peptide （cGRP） by immunohistochemistry. In addition， fluorescence dual-labeling was employed to determine the expression levels of CXCL16， the dendritic cell marker CD11c， the macrophage marker CD68， the microglia marker TMEM119， the endothelial cell markers CD31 and CXCR6， and the T cell marker CD3.Result Compared with the Sham group， the mechanical pain threshold of the CPSP group was significantly lower than that of the Sham group from day 3 to day 17， with stable hyperalgesia symptoms. On the 7th day， the mechanical pain threshold of the PGB group was significantly higher than that of the CPSP group， with significant analgesic effect （P<0.01）. On days 10-17, the mechanical pain threshold of the RYZBW-H group was significantly higher than that of the CPSP group， showing a stable analgesic effect （P<0.05）. On the 17th day， the analgesic effect of RYZBW was dose-effect correlated （R2=0.303 7）. From day 4 to day 17， the mechanical pain threshold of RYZBW-H group was positively correlated with time （R2=0.111 5）. The above results suggested that the analgesia of RYZBW was time-dependent. On the 17 th day， the expression of central sensitization marker cGRP in the spinal dorsal horn of CPSP mice was significantly increased compared with the Sham group （P<0.05）， and RYZBW down-regulated it in a dose-dependent manner （R2=0.500 8）， suggesting that RYZBW significantly inhibited the central sensitization of the spinal cord caused by CPSP. The results of spinal cord inflammation chip on the 17th day showed that compared with CPSP group， RYZBW-H group inhibited CXCL16 expression （P<0.01）.The results of ELISA based on independent repeated samples showed that RYZBW inhibited the expression of CXCL16 protein in spinal cord in a dose-dependent manner （R2=0.250 4）. The results of immunofluorescence double labeling showed that compared with Sham group, the expression of CXCL16 in CD11c positive dendritic cells in CPSP group increased， and the number of CD68 positive cells increased （P<0.05）. Compared with CPSP group， RYZBW down-regulated it: the expression of CXCL16 in CD31 positive endothelial cells， CD68 positive macrophages and TMEM119 positive microglia increased， and the number and cell body area of TMEM119 positive microglia increased significantly （P<0.05）. The number of CD3 positive T cells （P<0.05） and the expression of CXCR6 in CD3 positive T cells were increased. RYZBW inhibited the activation of endothelial cells and macrophages in a dose-dependent manner， and reduced the infiltration of microglia and T cells （R2=0.691 4， R2=0.551 5， R2=0.653 2， R2=0.180 6， R2=0.287 5， R2=0.298 6，R2=0.511 6）.Conclusion RYZBW can effectively alleviate nociceptive hypersensitivity and central sensitisation of the spinal cord in CPSP mice by regulating CXCL16-CXCR-6， inhibiting the infiltration and activation of microglia and macrophages， and the activation of dendritic cells， endothelial cells， and T cells.
Objective To clarify the pharmacodynamic characteristics and neuroinflammatory mechanisms of Ruyi Zhenbaowan （RYZBW） in treating nociceptive hypersensitivity and central sensitisation of spinal cord in the mouse model of central post-stroke pain （CPSP）.Method SPF-grade male ICR mice of 8 weeks old were assigned into the sham operation （Sham）， model （CPSP）， low-， medium-， and high-dose （0.303， 0.607 1.214 g·kg-1） RYZBW （RYZBW-L， RYZBW-M， and RYZBW-H， respectively）， and pregabalin （PGB， 0.046 g·kg-1， positive control） groups. The rat model of CPSP was established by injection of type Ⅳ collagenase into the ventral posterior lateral nucleus of the thalamus on day 1. Rats were administrated with corresponding drugs or normal saline （Sham and CPSP groups） by gavage from day 14 to day 17. The mechanical pain sensitivity test was performed on days 0， 3， 4， 7， 10， 14， 17. On day 18， the L5 segment of spinal cord was collected for the detection of inflammatory cytokines by immunoinflammatory microarray， CXC chemokine ligand 16 （CXCL16） by enzyme-linked immunosorbent assay， and calcitonin gene-related peptide （cGRP） by immunohistochemistry. In addition， fluorescence dual-labeling was employed to determine the expression levels of CXCL16， the dendritic cell marker CD11c， the macrophage marker CD68， the microglia marker TMEM119， the endothelial cell markers CD31 and CXCR6， and the T cell marker CD3.Result Compared with the Sham group， the mechanical pain threshold of the CPSP group was significantly lower than that of the Sham group from day 3 to day 17， with stable hyperalgesia symptoms. On the 7th day， the mechanical pain threshold of the PGB group was significantly higher than that of the CPSP group， with significant analgesic effect （P<0.01）. On days 10-17, the mechanical pain threshold of the RYZBW-H group was significantly higher than that of the CPSP group， showing a stable analgesic effect （P<0.05）. On the 17th day， the analgesic effect of RYZBW was dose-effect correlated （R2=0.303 7）. From day 4 to day 17， the mechanical pain threshold of RYZBW-H group was positively correlated with time （R2=0.111 5）. The above results suggested that the analgesia of RYZBW was time-dependent. On the 17 th day， the expression of central sensitization marker cGRP in the spinal dorsal horn of CPSP mice was significantly increased compared with the Sham group （P<0.05）， and RYZBW down-regulated it in a dose-dependent manner （R2=0.500 8）， suggesting that RYZBW significantly inhibited the central sensitization of the spinal cord caused by CPSP. The results of spinal cord inflammation chip on the 17th day showed that compared with CPSP group， RYZBW-H group inhibited CXCL16 expression （P<0.01）.The results of ELISA based on independent repeated samples showed that RYZBW inhibited the expression of CXCL16 protein in spinal cord in a dose-dependent manner （R2=0.250 4）. The results of immunofluorescence double labeling showed that compared with Sham group, the expression of CXCL16 in CD11c positive dendritic cells in CPSP group increased， and the number of CD68 positive cells increased （P<0.05）. Compared with CPSP group， RYZBW down-regulated it: the expression of CXCL16 in CD31 positive endothelial cells， CD68 positive macrophages and TMEM119 positive microglia increased， and the number and cell body area of TMEM119 positive microglia increased significantly （P<0.05）. The number of CD3 positive T cells （P<0.05） and the expression of CXCR6 in CD3 positive T cells were increased. RYZBW inhibited the activation of endothelial cells and macrophages in a dose-dependent manner， and reduced the infiltration of microglia and T cells （R2=0.691 4， R2=0.551 5， R2=0.653 2， R2=0.180 6， R2=0.287 5， R2=0.298 6，R2=0.511 6）.Conclusion RYZBW can effectively alleviate nociceptive hypersensitivity and central sensitisation of the spinal cord in CPSP mice by regulating CXCL16-CXCR-6， inhibiting the infiltration and activation of microglia and macrophages， and the activation of dendritic cells， endothelial cells， and T cells.
2024,30(24):47-56, DOI: 10.13422/j.cnki.syfjx.20240644
Abstract:
Objective To evaluate the intervention effect of Ruyi Zhenbaowan （RYZBW） on secondary brain injury and central pain in mice with hemorrhagic stroke and to explore its pharmacological mechanism of repairing the neurovascular unit from the perspective of neuroinflammation.Method A mouse model of central post-stroke pain （CPSP） was established by microinjecting type Ⅳ collagenase into the ventroposterior thalamic nucleus. The day of model establishment was recorded as D1， and the mice were divided into Sham operation group （Sham）， model group （CPSP）， low （RYZBW-L）， medium （RYZBW-M）， and high （RYZBW-H） dose groups of RYZBW， and positive drug pregabalin （PGB） group. On the 4th day （D4） after model establishment， gavage administration was performed twice daily. The Sham and CPSP groups received an equal volume of normal saline， while the RYZBW-L， RYZBW-M， and RYZBW-H groups received RYZBW at 1.214， 1.821， 2.428 g·kg-1， respectively， and the PGB group received PGB at 0.046 g·kg-1. Mechanical hyperalgesia was assessed before model establishment （D0）， on the 3rd day （D3）， and after the first gavage on D4. Nerve damage was evaluated after the second gavage on D1 and D4. On D4， peripheral blood was collected for routine blood tests， and the thalamus was collected for immune-inflammation microarray analysis. In independent samples， quantitative analysis was performed on the localization of immune-inflammatory factors， receptors， and cells via immunofluorescence staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot analysis.Result Compared with the Sham group， CPSP mice showed significant secondary nerve injury， central pain after stroke （P<0.05，P<0.01）， increased red blood cell distribution width （RDW） in peripheral blood （P<0.05）， and decreased hemoglobin （HGB） concentration （P<0.05）. Immune-inflammation microarray analysis showed that CC chemokine ligand 2 （CCL2） in the CPSP thalamus was significantly increased compared to the Sham group （P<0.01）， while CX3C chemokine ligand 1 （CX3CL1） was significantly decreased （P<0.05）. These results were confirmed by ELISA and immunofluorescence staining. Western blot analysis indicated that the protein expression of CX3CR1， the receptor for CX3CL1， was significantly decreased in the CPSP group compared to the Sham group （P<0.01）. Immunofluorescence staining revealed that the number of Ly6C+CX3CR1+ non-classical monocytes in the CPSP group did not change significantly， while the number of classical monocytes （CX3CR1-Ly6C+） significantly increased （P<0.01）. The expression of CX3CR1 in microglia was significantly increased in the CPSP group （P<0.01）. Compared with the CPSP group， RYZBW improved neurological deficits （R2=0.367 9） and central pain symptoms （R2=0.501 9） in a dose-dependent manner. RYZBW-H significantly improved peripheral blood RDW and HGB （P<0.05）. Immune-inflammation microarray analysis and ELISA results showed that RYZBW-H significantly inhibited CCL2 expression （P<0.01） and increased CX3CL1 expression （P<0.05）. Western blot results indicated that the protein expression of CX3CR1 in the RYZBW-L and RYZBW-H groups was significantly increased （P<0.05）. Immunofluorescence staining demonstrated that RYZBW increased the overall expression of CX3CR1 in a dose-dependent manner （R2=0.619 6）， inhibited the expression of CX3CR1 on microglia， and decreased both the number （R2=0.494 5） and soma area （R2=0.571 7） of microglia compared with the CPSP group. Additionally， RYZBW increased the infiltration of CX3CR1+Ly6C+ non-classical monocytes in a dose-dependent manner （R2=0.635 3） and effectively inhibited the infiltration of Ly6C+CX3CR1- classical monocytes （R2=0.483 6）.Conclusion RYZBW can effectively alleviate secondary injury and central pain in CPSP mice， and its mechanism involves regulating the CX3CL1-CX3CR1 ligand-receptor interaction， inhibiting microglial infiltration and activation， promoting non-classical monocyte infiltration for vascular repair， and suppressing the infiltration of classical monocytes for inflammatory phagocytosis.
Objective To evaluate the intervention effect of Ruyi Zhenbaowan （RYZBW） on secondary brain injury and central pain in mice with hemorrhagic stroke and to explore its pharmacological mechanism of repairing the neurovascular unit from the perspective of neuroinflammation.Method A mouse model of central post-stroke pain （CPSP） was established by microinjecting type Ⅳ collagenase into the ventroposterior thalamic nucleus. The day of model establishment was recorded as D1， and the mice were divided into Sham operation group （Sham）， model group （CPSP）， low （RYZBW-L）， medium （RYZBW-M）， and high （RYZBW-H） dose groups of RYZBW， and positive drug pregabalin （PGB） group. On the 4th day （D4） after model establishment， gavage administration was performed twice daily. The Sham and CPSP groups received an equal volume of normal saline， while the RYZBW-L， RYZBW-M， and RYZBW-H groups received RYZBW at 1.214， 1.821， 2.428 g·kg-1， respectively， and the PGB group received PGB at 0.046 g·kg-1. Mechanical hyperalgesia was assessed before model establishment （D0）， on the 3rd day （D3）， and after the first gavage on D4. Nerve damage was evaluated after the second gavage on D1 and D4. On D4， peripheral blood was collected for routine blood tests， and the thalamus was collected for immune-inflammation microarray analysis. In independent samples， quantitative analysis was performed on the localization of immune-inflammatory factors， receptors， and cells via immunofluorescence staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot analysis.Result Compared with the Sham group， CPSP mice showed significant secondary nerve injury， central pain after stroke （P<0.05，P<0.01）， increased red blood cell distribution width （RDW） in peripheral blood （P<0.05）， and decreased hemoglobin （HGB） concentration （P<0.05）. Immune-inflammation microarray analysis showed that CC chemokine ligand 2 （CCL2） in the CPSP thalamus was significantly increased compared to the Sham group （P<0.01）， while CX3C chemokine ligand 1 （CX3CL1） was significantly decreased （P<0.05）. These results were confirmed by ELISA and immunofluorescence staining. Western blot analysis indicated that the protein expression of CX3CR1， the receptor for CX3CL1， was significantly decreased in the CPSP group compared to the Sham group （P<0.01）. Immunofluorescence staining revealed that the number of Ly6C+CX3CR1+ non-classical monocytes in the CPSP group did not change significantly， while the number of classical monocytes （CX3CR1-Ly6C+） significantly increased （P<0.01）. The expression of CX3CR1 in microglia was significantly increased in the CPSP group （P<0.01）. Compared with the CPSP group， RYZBW improved neurological deficits （R2=0.367 9） and central pain symptoms （R2=0.501 9） in a dose-dependent manner. RYZBW-H significantly improved peripheral blood RDW and HGB （P<0.05）. Immune-inflammation microarray analysis and ELISA results showed that RYZBW-H significantly inhibited CCL2 expression （P<0.01） and increased CX3CL1 expression （P<0.05）. Western blot results indicated that the protein expression of CX3CR1 in the RYZBW-L and RYZBW-H groups was significantly increased （P<0.05）. Immunofluorescence staining demonstrated that RYZBW increased the overall expression of CX3CR1 in a dose-dependent manner （R2=0.619 6）， inhibited the expression of CX3CR1 on microglia， and decreased both the number （R2=0.494 5） and soma area （R2=0.571 7） of microglia compared with the CPSP group. Additionally， RYZBW increased the infiltration of CX3CR1+Ly6C+ non-classical monocytes in a dose-dependent manner （R2=0.635 3） and effectively inhibited the infiltration of Ly6C+CX3CR1- classical monocytes （R2=0.483 6）.Conclusion RYZBW can effectively alleviate secondary injury and central pain in CPSP mice， and its mechanism involves regulating the CX3CL1-CX3CR1 ligand-receptor interaction， inhibiting microglial infiltration and activation， promoting non-classical monocyte infiltration for vascular repair， and suppressing the infiltration of classical monocytes for inflammatory phagocytosis.
SUN Chunquan, XIE Yanming, GAO Jinghua, CHEN Weiheng, WANG Lianxin, WANG Shangquan, TIAN Xiangdong, XU Zujian, ZHENG Yuxin, ZHOU Mingwang, LI Chungen, XU Zhanwang, GUO Jiayi, DU Shuangqing, CHEN Qigang, JI Quan, BAI Zhiqiang, XIAO Jing, QI Wanli, YANG Weiyi, ZHANG Jingxiao
2024,30(24):57-67, DOI: 10.13422/j.cnki.syfjx.20241693
Abstract:
Objective This study aimed to evaluate the clinical efficacy of Ruyi Zhenbaowan（RYZBW）in the treatment of initial and early knee osteoarthritis （KOA） through a prospective multicenter，randomized，double-blind，and placebo-parallel controlled trial.Method From October 13th， 2021 to December 25th， 2021， 240 KOA subjects meeting the acceptance criteria were enrolled in 15 sub-centers including Wangjing Hospital， Chinese Academy of Chinese Medical Sciences， and they were randomly divided into observation group and control group， with 120 cases in each group. The intervention measures for the observation group were RYZBW + health education， and the intervention measures for the control group were RYZBW placebo + health education. The intervention period in both groups was four weeks， and they were followed up for four weeks after the intervention. The primary outcome measure was the total score of Western Ontario and McMaster University Osteoarthritis Index score （WOMAC score）， and the secondary outcome measures were the response rate of visual scale （VAS） pain score， WOMAC sub item scores （joint pain， joint stiffness， and joint function）， quality of life （SF-12） score， and traditional Chinese medicine （TCM） syndrome score.Result （1） Efficacy evaluation. The marginal model results showed that the observation group was better than the control group in improving the WOMAC total score and WOMAC pain score in the treatment of KOA with RYZBW， and the difference was statistically significant （P<0.05）. There was no significant difference between the two groups in improving VAS score response rate， WOMAC function score， WOMAC stiffness score， SF12-PCS （quality of life-physical health） score， SF12-MCS （quality of life-mental health） score， and TCM syndrome score. （2） Subgroup analysis. ① In terms of VAS score response rate， the response rate of the observation group was higher than that of the control group for subjects with baseline VAS score of （4， 5］， and the difference was statistically significant （P<0.05）. ② In terms of TCM syndrome score， for subjects aged ［56， 60］ and ［61， 65］， the decrease in total TCM syndrome score in the observation group was better than that in the control group， and the difference was statistically significant （P<0.05）.Conclusion Tibetan medicine RYZBW has good clinical efficacy in improving WOMAC total score， VAS score response rate， WOMAC pain score， WOMAC function score， and TCM syndrome score for patients with initial and early KOA， which can fill the lack of Tibetan medicine RYZBW in the treatment of KOA and make a demonstration study for the inheritance and development of ethnic medicine.
Objective This study aimed to evaluate the clinical efficacy of Ruyi Zhenbaowan（RYZBW）in the treatment of initial and early knee osteoarthritis （KOA） through a prospective multicenter，randomized，double-blind，and placebo-parallel controlled trial.Method From October 13th， 2021 to December 25th， 2021， 240 KOA subjects meeting the acceptance criteria were enrolled in 15 sub-centers including Wangjing Hospital， Chinese Academy of Chinese Medical Sciences， and they were randomly divided into observation group and control group， with 120 cases in each group. The intervention measures for the observation group were RYZBW + health education， and the intervention measures for the control group were RYZBW placebo + health education. The intervention period in both groups was four weeks， and they were followed up for four weeks after the intervention. The primary outcome measure was the total score of Western Ontario and McMaster University Osteoarthritis Index score （WOMAC score）， and the secondary outcome measures were the response rate of visual scale （VAS） pain score， WOMAC sub item scores （joint pain， joint stiffness， and joint function）， quality of life （SF-12） score， and traditional Chinese medicine （TCM） syndrome score.Result （1） Efficacy evaluation. The marginal model results showed that the observation group was better than the control group in improving the WOMAC total score and WOMAC pain score in the treatment of KOA with RYZBW， and the difference was statistically significant （P<0.05）. There was no significant difference between the two groups in improving VAS score response rate， WOMAC function score， WOMAC stiffness score， SF12-PCS （quality of life-physical health） score， SF12-MCS （quality of life-mental health） score， and TCM syndrome score. （2） Subgroup analysis. ① In terms of VAS score response rate， the response rate of the observation group was higher than that of the control group for subjects with baseline VAS score of （4， 5］， and the difference was statistically significant （P<0.05）. ② In terms of TCM syndrome score， for subjects aged ［56， 60］ and ［61， 65］， the decrease in total TCM syndrome score in the observation group was better than that in the control group， and the difference was statistically significant （P<0.05）.Conclusion Tibetan medicine RYZBW has good clinical efficacy in improving WOMAC total score， VAS score response rate， WOMAC pain score， WOMAC function score， and TCM syndrome score for patients with initial and early KOA， which can fill the lack of Tibetan medicine RYZBW in the treatment of KOA and make a demonstration study for the inheritance and development of ethnic medicine.
XU Mingzhu, LI Huaiping, MA Zhaochen, LI Tao, LIU Yudong, XIAO Ziqing, ZHANG Chu, CHEN Kedian, MA Weihua, HUANG Feng, LIN Na, ZHANG Yanqiong
2024,30(24):68-77, DOI: 10.13422/j.cnki.syfjx.20250261
Abstract:
Objective To identify the pharmacodynamic material basis of Ruyi Zhenbaowan in relieving neuropathic pain by integrating the calculation of biological network proximity and pharmacokinetic characterization.Method The interaction network of "drug candidate target-related gene of disease" was constructed by Cytoscape 3.8.2， and the average shortest path value of each drug putative target acting on neuropathic pain-related genes in this network was calculated by Pesca 3.8.0 tool so as to evaluate the network proximity between them， and screen prescription candidate targets with strong intervention efficiency and their corresponding potential effect components. After that， plasma and cerebrospinal fluid samples were collected from rats after administration of Ruyi Zhenbaowan at set time points， and the contents of potential effect components in samples was quantified by ultra performance liquid chromatography-quadrupole-ion trap mass spectrometry（UPLC-Q-TRAP/MS）， and drug concentration-time curves were plotted， then the pharmacokinetic parameters were calculated by DAS 2.1.1.Result By evaluating the network proximity between candidate targets and neuropathic pain-related genes in the interaction network， a total of 40 putative targets of Ruyi Zhenbaowan with strong intervention effects on neuropathic pain-related genes， such as estrogen receptor 1（ESR1）， cyclic adenosine monophosphate（cAMP）-dependent protein kinase catalytic subunit alpha（PRKACA） and protein kinase B1 （Akt1）， and 10 corresponding potential effect components， such as glycyrrhizic acid and betulinic acid， were obtained. Pharmacokinetic characterization showed that among the 10 potential effect components， gallic acid， apigenin-7-O-glucuronide， glycyrrhizic acid and apigenin were well absorbed and metabolized in plasma and cerebrospinal fluid， with long onset time and good bioavailability.Conclusion From the perspective of efficacy-target-constituent-pharmacokinetic， this study analyzes the main effective materials of Ruyi Zhenbaowan， such as glycyrrhizic acid， gallic acid， apigenin-7-O-glucuronide and apigenin， which have a high exposure in plasma or cerebrospinal fluid and have a strong intervention effect on neuropathic pain. The related results provide reliable experimental evidences for clarifying the material basis and developing quality standards of Ruyi Zhenbaowan.
Objective To identify the pharmacodynamic material basis of Ruyi Zhenbaowan in relieving neuropathic pain by integrating the calculation of biological network proximity and pharmacokinetic characterization.Method The interaction network of "drug candidate target-related gene of disease" was constructed by Cytoscape 3.8.2， and the average shortest path value of each drug putative target acting on neuropathic pain-related genes in this network was calculated by Pesca 3.8.0 tool so as to evaluate the network proximity between them， and screen prescription candidate targets with strong intervention efficiency and their corresponding potential effect components. After that， plasma and cerebrospinal fluid samples were collected from rats after administration of Ruyi Zhenbaowan at set time points， and the contents of potential effect components in samples was quantified by ultra performance liquid chromatography-quadrupole-ion trap mass spectrometry（UPLC-Q-TRAP/MS）， and drug concentration-time curves were plotted， then the pharmacokinetic parameters were calculated by DAS 2.1.1.Result By evaluating the network proximity between candidate targets and neuropathic pain-related genes in the interaction network， a total of 40 putative targets of Ruyi Zhenbaowan with strong intervention effects on neuropathic pain-related genes， such as estrogen receptor 1（ESR1）， cyclic adenosine monophosphate（cAMP）-dependent protein kinase catalytic subunit alpha（PRKACA） and protein kinase B1 （Akt1）， and 10 corresponding potential effect components， such as glycyrrhizic acid and betulinic acid， were obtained. Pharmacokinetic characterization showed that among the 10 potential effect components， gallic acid， apigenin-7-O-glucuronide， glycyrrhizic acid and apigenin were well absorbed and metabolized in plasma and cerebrospinal fluid， with long onset time and good bioavailability.Conclusion From the perspective of efficacy-target-constituent-pharmacokinetic， this study analyzes the main effective materials of Ruyi Zhenbaowan， such as glycyrrhizic acid， gallic acid， apigenin-7-O-glucuronide and apigenin， which have a high exposure in plasma or cerebrospinal fluid and have a strong intervention effect on neuropathic pain. The related results provide reliable experimental evidences for clarifying the material basis and developing quality standards of Ruyi Zhenbaowan.
CHEN Kedian, MA Zhaochen, CAI Bingbing, LIU Ying, LIU Yudong, LI Tao, XU Mingzhu, WANG Haiping, LIN Na, ZHANG Yanqiong
2024,30(24):78-84, DOI: 10.13422/j.cnki.syfjx.20241443
Abstract:
Objective To identify the chemical constituents of Ruyi Zhenbaowan in vitro and in vivo.Method The chemical constituents of Ruyi Zhenbaowan were identified based on UHPLC-Q Exactive Orbitrap HRMS. A total of 12 male SD rats were randomized into two groups： control （pure water） and Ruyi Zhenbaowan （1.8 g·kg-1）. The rats were administrated with the suspension of Ruyi Zhenbaowan or pure water by gavage. After 1.5 h， the plasma and cerebrospinal fluid were collected. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C18 column （2.1 mm × 150 mm， 1.7 μm） with a mixture of 0.1% formic acid aqueous solution （A） and acetonitrile （B） as the mobile phase. Gradient elution was carried out according to the procedure of 0~15 min，97%~80%A；15~30 min ，80%~60%A；30~40 min，60%~30%A；40~45 min，30%~5%A. The ion source was electrospray ionization， and scan range was m/z 100-1 500. The prototype components and the components in the plasma and cerebrospinal fluid were analyzed qualitatively by scanning in positive and negative ion modes and identified by comparison with the data in published literature and the information of standard substances.Result A total of 126 chemical constituents were identified from the 80% methanol solution of Ruyi Zhenbaowan， and 14 and 7 prototype constituents were detected in the plasma and the cerebrospinal fluid， respectively. In addition， the fragmentation rules of apigenin， apigenin-7-O-glucuronide， galangin， liquiritin， piperine， glycyrrhizic acid， eugenol， gallic acid， and cholic acid were deduced.Conclusion This study achieved rapid multicomponent characterization and identification of Ruyi Zhenbaowan in vitro and in vivo， providing theoretical support for exploring active substances and performing quality control.l.
Objective To identify the chemical constituents of Ruyi Zhenbaowan in vitro and in vivo.Method The chemical constituents of Ruyi Zhenbaowan were identified based on UHPLC-Q Exactive Orbitrap HRMS. A total of 12 male SD rats were randomized into two groups： control （pure water） and Ruyi Zhenbaowan （1.8 g·kg-1）. The rats were administrated with the suspension of Ruyi Zhenbaowan or pure water by gavage. After 1.5 h， the plasma and cerebrospinal fluid were collected. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C18 column （2.1 mm × 150 mm， 1.7 μm） with a mixture of 0.1% formic acid aqueous solution （A） and acetonitrile （B） as the mobile phase. Gradient elution was carried out according to the procedure of 0~15 min，97%~80%A；15~30 min ，80%~60%A；30~40 min，60%~30%A；40~45 min，30%~5%A. The ion source was electrospray ionization， and scan range was m/z 100-1 500. The prototype components and the components in the plasma and cerebrospinal fluid were analyzed qualitatively by scanning in positive and negative ion modes and identified by comparison with the data in published literature and the information of standard substances.Result A total of 126 chemical constituents were identified from the 80% methanol solution of Ruyi Zhenbaowan， and 14 and 7 prototype constituents were detected in the plasma and the cerebrospinal fluid， respectively. In addition， the fragmentation rules of apigenin， apigenin-7-O-glucuronide， galangin， liquiritin， piperine， glycyrrhizic acid， eugenol， gallic acid， and cholic acid were deduced.Conclusion This study achieved rapid multicomponent characterization and identification of Ruyi Zhenbaowan in vitro and in vivo， providing theoretical support for exploring active substances and performing quality control.l.
TIAN Shiqiu, ZUO Zeping, TIAN Yingying, LI Yilin, PEI Hailuan, LIN Zhaozhou, LYU Yingnan, WANG Jianfang, WANG Zhibin
2024,30(24):85-94, DOI: 10.13422/j.cnki.syfjx.20241404
Abstract:
Objective To explore the therapeutic effect and mechanism of the Danhe granules on hypercholesterolemia rats by observing the changes in the efficacy indicators and the levels of proteins related to the cholesterol metabolism pathway in the rats under the intervention of Danhe granules.Method SD rats were randomly assigned to either the blank group or the model group based on their body weight. The blank group had normal chow diets， while the model group was fed high-fat diets for seven weeks. One week after the establishment of the model， the content of the serum total cholesterol （TC） in the model rats was detected. According to the TC value， the model group was further randomly divided into a control group， pravastatin sodium tablet group（4.02 mg·kg-1）， Xuezhikang capsule group（0.12 g·kg-1）， high-dose， middle-dose， and low-dose groups of Danhe granules（4.536， 2.268， 1.134 g·kg-1）. After grouping the model groups， each treatment group received continuous oral gavage for six weeks， with weekly measurements of body weight and food intake （the difference between feed intake and feed surplus）. Six weeks later， the levels of serum TC， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） were measured. The liver pathology and lipid droplet distribution were evaluated by hematoxylin-eosin （HE） staining and oil red O staining， with scoring and calculation conducted. Rat liver tissue was collected， and western blot and immunohistochemistry （IHC） were used to detect the expression levels of cholesterol metabolism-related proteins namely phosphorylated adenosine 5'-monophosphate （AMP）-activated protein kinase （p-AMPK）， AMPK， 3-hydroxy-3-methyl glutaryl coenzyme A reductase （HMGCR）， low-density lipoprotein receptor （LDLR）， cholesterol 7α-hydroxylase （CYP7A1）， Acyl-coenzyme A： cholesterol acyltransferase 2 （ACAT2）， and apolipoprotein B （ApoB） in hypercholesterolemia rats.Result Compared with the blank group， the model group showed a significantly higher level of serum TC （P<0.01）. The TG level had no significant change， and the HDL-C level was significantly decreased （P<0.05）. The liver index， steatosis score， total score of pathological state， and the positive area ratio of oil red O staining were significantly increased （P<0.01）， and the protein expression levels of p-AMPK， p-AMPK/AMPK， LDLR， and CYP7A1 were significantly decreased （P<0.05， P<0.01）， while the protein expression levels of AMPK， HMGCR， and ACAT2 were significantly increased （P<0.05， P<0.01）. Compared with the model group， the TC level in each dose group of Danhe granules was significantly decreased （P<0.05）， and the positive area ratio of oil red O staining in the pravastatin sodium tablet group and medium-dose group of Danhe granules was significantly decreased （P<0.05）. In each administration group， the protein expression levels of p-AMPK and p-AMPK/AMPK were significantly increased （P<0.05， P<0.01）， and the levels of HMGCR and ACAT2 were significantly decreased （P<0.01）. The ApoB level showed a downward trend. The CYP7A1 level in the pravastatin sodium tablet group and each dose group of Danhe granules was significantly increased （P<0.05， P<0.01）， and the LDLR level in the pravastatin sodium tablet group， Xuezhikang capsule group， and high-dose and medium-dose groups of Danhe granules was significantly increased （P<0.05， P<0.01）.Conclusion Danhe granules can reduce serum TC levels and improve hepatic steatosis. It may activate AMPK， down-regulate the expression of HMGCR， and inhibit cholesterol synthesis. It can also up-regulate the expression of LDLR and CYP7A1， promote cholesterol uptake and excretion， down-regulate the expression of ACAT2 and ApoB， reduce cholesterol absorption and assembly of LDL and other lipoproteins， and thus play a role in the treatment of hypercholesterolemia.
Objective To explore the therapeutic effect and mechanism of the Danhe granules on hypercholesterolemia rats by observing the changes in the efficacy indicators and the levels of proteins related to the cholesterol metabolism pathway in the rats under the intervention of Danhe granules.Method SD rats were randomly assigned to either the blank group or the model group based on their body weight. The blank group had normal chow diets， while the model group was fed high-fat diets for seven weeks. One week after the establishment of the model， the content of the serum total cholesterol （TC） in the model rats was detected. According to the TC value， the model group was further randomly divided into a control group， pravastatin sodium tablet group（4.02 mg·kg-1）， Xuezhikang capsule group（0.12 g·kg-1）， high-dose， middle-dose， and low-dose groups of Danhe granules（4.536， 2.268， 1.134 g·kg-1）. After grouping the model groups， each treatment group received continuous oral gavage for six weeks， with weekly measurements of body weight and food intake （the difference between feed intake and feed surplus）. Six weeks later， the levels of serum TC， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） were measured. The liver pathology and lipid droplet distribution were evaluated by hematoxylin-eosin （HE） staining and oil red O staining， with scoring and calculation conducted. Rat liver tissue was collected， and western blot and immunohistochemistry （IHC） were used to detect the expression levels of cholesterol metabolism-related proteins namely phosphorylated adenosine 5'-monophosphate （AMP）-activated protein kinase （p-AMPK）， AMPK， 3-hydroxy-3-methyl glutaryl coenzyme A reductase （HMGCR）， low-density lipoprotein receptor （LDLR）， cholesterol 7α-hydroxylase （CYP7A1）， Acyl-coenzyme A： cholesterol acyltransferase 2 （ACAT2）， and apolipoprotein B （ApoB） in hypercholesterolemia rats.Result Compared with the blank group， the model group showed a significantly higher level of serum TC （P<0.01）. The TG level had no significant change， and the HDL-C level was significantly decreased （P<0.05）. The liver index， steatosis score， total score of pathological state， and the positive area ratio of oil red O staining were significantly increased （P<0.01）， and the protein expression levels of p-AMPK， p-AMPK/AMPK， LDLR， and CYP7A1 were significantly decreased （P<0.05， P<0.01）， while the protein expression levels of AMPK， HMGCR， and ACAT2 were significantly increased （P<0.05， P<0.01）. Compared with the model group， the TC level in each dose group of Danhe granules was significantly decreased （P<0.05）， and the positive area ratio of oil red O staining in the pravastatin sodium tablet group and medium-dose group of Danhe granules was significantly decreased （P<0.05）. In each administration group， the protein expression levels of p-AMPK and p-AMPK/AMPK were significantly increased （P<0.05， P<0.01）， and the levels of HMGCR and ACAT2 were significantly decreased （P<0.01）. The ApoB level showed a downward trend. The CYP7A1 level in the pravastatin sodium tablet group and each dose group of Danhe granules was significantly increased （P<0.05， P<0.01）， and the LDLR level in the pravastatin sodium tablet group， Xuezhikang capsule group， and high-dose and medium-dose groups of Danhe granules was significantly increased （P<0.05， P<0.01）.Conclusion Danhe granules can reduce serum TC levels and improve hepatic steatosis. It may activate AMPK， down-regulate the expression of HMGCR， and inhibit cholesterol synthesis. It can also up-regulate the expression of LDLR and CYP7A1， promote cholesterol uptake and excretion， down-regulate the expression of ACAT2 and ApoB， reduce cholesterol absorption and assembly of LDL and other lipoproteins， and thus play a role in the treatment of hypercholesterolemia.
YUAN Yulu, HE Zhanzhan, CHU Ce, TAO Xuguang, YANG Zhen, CHEN Xiangyun, DING Wei, XU Yongqi, ZHANG Yuxin, ZHAO Peizhang, CHEN Wanping, ZHAO Hongxia, WANG Wenlai
2024,30(24):95-102, DOI: 10.13422/j.cnki.syfjx.20241501
Abstract:
Objective To investigate the mechanism and effect of Qingfei Paidu decoction on transient receptor potential vanilloid-1/Transient receptor potential ankyrin1 （TRPV1/TRPA1） based on heat-sensitive channel and inflammatory response.Method According to body weight， 80 8-week-old C57BL/6 mice were randomly divided into the normal group， model group， dexamethasone group （5 mg·kg-1）， and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction （14.865， 29.73， 59.46 g·kg-1）， with 12 mice in each group. In addition to the normal group， the other groups were administered 20 μL （1×10-3 g·kg-1） to each mouse by airway infusion to establish the acute lung injury （ALI） model. In the administration group， the drug was given 1 h after modeling and again after an interval of 24 h. The lung tissue was taken 36 h after modeling. Double lung wet/dry weight ratio（W/D）， hematoxylin-eosin （HE） staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot were used to observe and detect the pathological changes of lung tissue， expression levels of inflammatory cytokine tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）， and expressions of TRPV1 and TRPA1 proteins in heat-sensitive channel， nuclear factor kappa-B （NF-κB）， inhibitor of NF-κB （IκBα） in inflammatory pathway， and phosphorylated proteins. The phosphorylated protein/total protein ratio was calculated.Result Compared with that in the normal group， the lung tissue of mice in the model group was seriously damaged， and pulmonary capillary permeability increased. Alveolar capillary congestion and dilation destroyed the complete structure of the alveolar， and the alveolar wall thickened. A large number of inflammatory cells and red blood cells were infiltrated， and pulmonary edema was significantly aggravated. The expressions of TNF-α， IL-6， TRPV1， TRPA1， phosphorylated NF-κB p65/NF-κB p65， and phosphorylated IκBα/IκBα were significantly increased （P<0.01）， and the whole lung W/D was significantly increased （P<0.01）. Compared with the model group， the dexamethasone group and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction could significantly improve pulmonary edema. TNF-α， IL-6， TRPV1， TRPA1， lung tissue NF-κB p65， and IκBα phosphorylated protein/total protein ratio decreased significantly （P<0.05， P<0.01）. The whole lung W/D also decreased significantly （P<0.05， P<0.01）.Conclusion Qingfei Paidu decoction has anti-inflammatory and protective effects on LPS-ALI mice， which can effectively reduce inflammation， induce diuresis， and alleviate edema. Its mechanism may be related to the regulation of the expression of TRPA1 and TRPV1 and the inhibition of the activation of the NF-κB pathway.
Objective To investigate the mechanism and effect of Qingfei Paidu decoction on transient receptor potential vanilloid-1/Transient receptor potential ankyrin1 （TRPV1/TRPA1） based on heat-sensitive channel and inflammatory response.Method According to body weight， 80 8-week-old C57BL/6 mice were randomly divided into the normal group， model group， dexamethasone group （5 mg·kg-1）， and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction （14.865， 29.73， 59.46 g·kg-1）， with 12 mice in each group. In addition to the normal group， the other groups were administered 20 μL （1×10-3 g·kg-1） to each mouse by airway infusion to establish the acute lung injury （ALI） model. In the administration group， the drug was given 1 h after modeling and again after an interval of 24 h. The lung tissue was taken 36 h after modeling. Double lung wet/dry weight ratio（W/D）， hematoxylin-eosin （HE） staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot were used to observe and detect the pathological changes of lung tissue， expression levels of inflammatory cytokine tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）， and expressions of TRPV1 and TRPA1 proteins in heat-sensitive channel， nuclear factor kappa-B （NF-κB）， inhibitor of NF-κB （IκBα） in inflammatory pathway， and phosphorylated proteins. The phosphorylated protein/total protein ratio was calculated.Result Compared with that in the normal group， the lung tissue of mice in the model group was seriously damaged， and pulmonary capillary permeability increased. Alveolar capillary congestion and dilation destroyed the complete structure of the alveolar， and the alveolar wall thickened. A large number of inflammatory cells and red blood cells were infiltrated， and pulmonary edema was significantly aggravated. The expressions of TNF-α， IL-6， TRPV1， TRPA1， phosphorylated NF-κB p65/NF-κB p65， and phosphorylated IκBα/IκBα were significantly increased （P<0.01）， and the whole lung W/D was significantly increased （P<0.01）. Compared with the model group， the dexamethasone group and low-dose， medium-dose， and high-dose groups of Qingfei Paidu decoction could significantly improve pulmonary edema. TNF-α， IL-6， TRPV1， TRPA1， lung tissue NF-κB p65， and IκBα phosphorylated protein/total protein ratio decreased significantly （P<0.05， P<0.01）. The whole lung W/D also decreased significantly （P<0.05， P<0.01）.Conclusion Qingfei Paidu decoction has anti-inflammatory and protective effects on LPS-ALI mice， which can effectively reduce inflammation， induce diuresis， and alleviate edema. Its mechanism may be related to the regulation of the expression of TRPA1 and TRPV1 and the inhibition of the activation of the NF-κB pathway.
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2013,19(20):246-250, DOI: 10.11653/syfj2013200246
Abstract:
Objective: To investigate the intervention effect and potential mechanism of Qingre Huashi decoction(QHD) for preventing the onset of colon cancer by observing SGC-7901 cell proliferation and apoptosis along with the expression of CD14 gene, tumor necrosis factor alpha and interleukin 1 beta. Method: SGC-7901 cells were divided into control group, intervention group. In vitro, the control group was offered fetal calf serum. The intervention group was fed with serum containing QHD for 5%,10%,15%,20%,30%. The multiplication, proliferation and apoptosis of SGC-7901 cell were detected by MTT and flow cytometry respectively. The expression of CD14, TNF-α and IL-1β gene was assayed by immunohistochemistry and real-time PCR. Result: Compared with control group, the survival of SGC-7901 cell was decreased in QHD group(P<0.05). In QHD group, the cells in G0/G1 phase were increased but the S phase cell was on the decline. Those changes were correlated with the culturing content. There is a significant difference between QHD group and control group in those changes as mentioned above(P<0.05). Especially, the effect of middle concentration serum containing QHD on inducing cell apoptosis and down regulating the expression of CD14, TNF-α, IL-1β was obvious (P<0.05). Conclusion: QHD exerts a distinct effect on preventing the onset of colon cancer by mediating SGC-7901 cell S time, cell composition and regulating the expression of CD14, TNF-α and IL-1β and inducing cell apoptosis.
Objective: To investigate the intervention effect and potential mechanism of Qingre Huashi decoction(QHD) for preventing the onset of colon cancer by observing SGC-7901 cell proliferation and apoptosis along with the expression of CD14 gene, tumor necrosis factor alpha and interleukin 1 beta. Method: SGC-7901 cells were divided into control group, intervention group. In vitro, the control group was offered fetal calf serum. The intervention group was fed with serum containing QHD for 5%,10%,15%,20%,30%. The multiplication, proliferation and apoptosis of SGC-7901 cell were detected by MTT and flow cytometry respectively. The expression of CD14, TNF-α and IL-1β gene was assayed by immunohistochemistry and real-time PCR. Result: Compared with control group, the survival of SGC-7901 cell was decreased in QHD group(P<0.05). In QHD group, the cells in G0/G1 phase were increased but the S phase cell was on the decline. Those changes were correlated with the culturing content. There is a significant difference between QHD group and control group in those changes as mentioned above(P<0.05). Especially, the effect of middle concentration serum containing QHD on inducing cell apoptosis and down regulating the expression of CD14, TNF-α, IL-1β was obvious (P<0.05). Conclusion: QHD exerts a distinct effect on preventing the onset of colon cancer by mediating SGC-7901 cell S time, cell composition and regulating the expression of CD14, TNF-α and IL-1β and inducing cell apoptosis.
2015,21(11):7-10, DOI: 10.13422/j.cnki.syfjx.2015110007
Abstract:
Objective: To investigate correlation between content and anti-proliferative effect against lung carcinoma cell line SPC-A-1 and A549 of active ingredients in Coicis Semen from five places. Method: HPLC-ELSD and UV spectrophotometry were employed to determine contents of glycerol trioleate and total triglycerides in Coicis Semen,respectively.In addition,MTT assay was performed to explore anti-proliferative effect of Coicis Semen extract from various origins towards SPC-A-1 and A549 cells. Result: The content of glycerol trioleate in Coicis Semen from Hebei,Liaoning,Fujian,Guizhou,Shandong province was 0.91%,1.14%,0.82%,0.77%,1.09%,the content of total triglycerides was 14.83,10.88,7.85,5.26,7.71 mg·g-1,respectively.IC50(median inhibitory concentration) against the lung carcinoma cell line A549 were 2.33,2.77,3.15,3.24,3.08 g·L-1,IC50 against SPC-A-1 were 1.64,2.15,2.63,3.01,2.99 g·L-1,respectively. Conclusion: All samples originated from five provinces are in line with standards of the 2010 edition of Chinese Pharmacopoeia.However,samples from Hebei province have the highest content of total triglycerides and the strongest anti-lung cancer activity,suggesting a positive correlation between active ingredients content and pharmacological activity.
Objective: To investigate correlation between content and anti-proliferative effect against lung carcinoma cell line SPC-A-1 and A549 of active ingredients in Coicis Semen from five places. Method: HPLC-ELSD and UV spectrophotometry were employed to determine contents of glycerol trioleate and total triglycerides in Coicis Semen,respectively.In addition,MTT assay was performed to explore anti-proliferative effect of Coicis Semen extract from various origins towards SPC-A-1 and A549 cells. Result: The content of glycerol trioleate in Coicis Semen from Hebei,Liaoning,Fujian,Guizhou,Shandong province was 0.91%,1.14%,0.82%,0.77%,1.09%,the content of total triglycerides was 14.83,10.88,7.85,5.26,7.71 mg·g-1,respectively.IC50(median inhibitory concentration) against the lung carcinoma cell line A549 were 2.33,2.77,3.15,3.24,3.08 g·L-1,IC50 against SPC-A-1 were 1.64,2.15,2.63,3.01,2.99 g·L-1,respectively. Conclusion: All samples originated from five provinces are in line with standards of the 2010 edition of Chinese Pharmacopoeia.However,samples from Hebei province have the highest content of total triglycerides and the strongest anti-lung cancer activity,suggesting a positive correlation between active ingredients content and pharmacological activity.
2014,20(5):54-56, DOI: 10.11653/syfj2014050054
Abstract:
Objective: To establish a high performance liquid chromatographic method (HPLC) for simultaneous determination of rubrofusarin gentiobioside, casside, cassiaside C in Cassia obtusifolia. Method: The HPLC with Dionex Acclaim C18(4.6 mm×250 mm, 5 μm) column was used, acetonitrile-THF-1% acetic acid (17 ：2 ：81) was used as a mobile phase, with flow rate of 1 mL·min-1, column temperature at 30 ℃ and detection wavelength at 278 nm. Result: Rubrofusarin gentiobioside, casside, cassiaside C showed good linearity (r=0.999 9) in the range of 0.218-1.09, 0.188-0.94, 0.200-1.000 μg, regressive equation were Y=54.017X-0.344 6 (r=0.999 9),Y=78.63X-0.354(r=0.999 9),Y=74.098X-0.433 5 (r=0.999 9)respectively;the average recoveries of the method were 98.43%, 101.89%, 102.46%, RSD were 1.75%, 1.10%, 1.15%. Conclusion: The method was proved to be simple, rapid, sensitive, precise, reliable and repeatable. It can be applied to the quality control of Semen Cassia.
Objective: To establish a high performance liquid chromatographic method (HPLC) for simultaneous determination of rubrofusarin gentiobioside, casside, cassiaside C in Cassia obtusifolia. Method: The HPLC with Dionex Acclaim C18(4.6 mm×250 mm, 5 μm) column was used, acetonitrile-THF-1% acetic acid (17 ：2 ：81) was used as a mobile phase, with flow rate of 1 mL·min-1, column temperature at 30 ℃ and detection wavelength at 278 nm. Result: Rubrofusarin gentiobioside, casside, cassiaside C showed good linearity (r=0.999 9) in the range of 0.218-1.09, 0.188-0.94, 0.200-1.000 μg, regressive equation were Y=54.017X-0.344 6 (r=0.999 9),Y=78.63X-0.354(r=0.999 9),Y=74.098X-0.433 5 (r=0.999 9)respectively;the average recoveries of the method were 98.43%, 101.89%, 102.46%, RSD were 1.75%, 1.10%, 1.15%. Conclusion: The method was proved to be simple, rapid, sensitive, precise, reliable and repeatable. It can be applied to the quality control of Semen Cassia.
2014,20(9):15-18, DOI: 10.13422/j.cnki.syfix.2014090015
Abstract:
Objective: To optimize extraction technology of Jiedu Tongluo particles. Method: Based on single factor tests, central composite design-response surface methodology was applied to optimize extraction process with soaking time, decoction time and solvent amounts as independent variables, while composite score of extraction amounts of baicalin, quercetin and isorhamnetin as dependent variable.SPSS software was used to fit multivariate linear equation and second-order polynomial equation for experimental data, response surface and contour plot were delineated according to best-fit mathematic models by Design Expert 8.0.0 statistic analysis software. Result: Optimum extraction process was as follows:soaking time 90 min, decoction time 85.63 min, the amount of solvent 9.21 times;Composite score was (93.2±0.051) %, which had deviation of -2.32% with the predicted value of 94.30%. Conclusion: Model established by central composite design-response surface methodology had good predictability, which was suitable for optimizing extraction process of Jiedu Tongluo particles.
Objective: To optimize extraction technology of Jiedu Tongluo particles. Method: Based on single factor tests, central composite design-response surface methodology was applied to optimize extraction process with soaking time, decoction time and solvent amounts as independent variables, while composite score of extraction amounts of baicalin, quercetin and isorhamnetin as dependent variable.SPSS software was used to fit multivariate linear equation and second-order polynomial equation for experimental data, response surface and contour plot were delineated according to best-fit mathematic models by Design Expert 8.0.0 statistic analysis software. Result: Optimum extraction process was as follows:soaking time 90 min, decoction time 85.63 min, the amount of solvent 9.21 times;Composite score was (93.2±0.051) %, which had deviation of -2.32% with the predicted value of 94.30%. Conclusion: Model established by central composite design-response surface methodology had good predictability, which was suitable for optimizing extraction process of Jiedu Tongluo particles.
Abstract:
【Abstract】Objective To study the mechanism of the chinese medicine Weichangshu on mitochondrial kinetic energy in diabetic mellitus rats stomach by measuring the changes of morphology, structure, function and protein of mitochondria. Methods: Standard Sprague Dawley rats with a mean weight of 150-170g were randomly divided into 4 groups, including a control group, DM model group, DM WeichangshuⅠgroup, DM Weichangshu Ⅱ group. The mitochondrial morphology and structure were detected using transmission electron microscopy. The changes of mitochondrial transmembrane potential, respiratory chain and mitochondrial protein were detected using flow cytometry, fluorescence labeling and Western Blot respectively. Results: Weichangshu could ameliorate the mitochondrial morphology, structure and respiratory chain in diabetic mellitus rats stomach. Mitochondrial transmembrane potential, ATP level and activity of cytochrome C oxidase were increased. The release of ROS was decreased. The expression of Bcl-2 up-regulated and bax down-regulated. Conclusion: Weichangshu could promoted the gastrointestinal motility, which might be related with the regulation of mitochondrial kinetic energy.
【Abstract】Objective To study the mechanism of the chinese medicine Weichangshu on mitochondrial kinetic energy in diabetic mellitus rats stomach by measuring the changes of morphology, structure, function and protein of mitochondria. Methods: Standard Sprague Dawley rats with a mean weight of 150-170g were randomly divided into 4 groups, including a control group, DM model group, DM WeichangshuⅠgroup, DM Weichangshu Ⅱ group. The mitochondrial morphology and structure were detected using transmission electron microscopy. The changes of mitochondrial transmembrane potential, respiratory chain and mitochondrial protein were detected using flow cytometry, fluorescence labeling and Western Blot respectively. Results: Weichangshu could ameliorate the mitochondrial morphology, structure and respiratory chain in diabetic mellitus rats stomach. Mitochondrial transmembrane potential, ATP level and activity of cytochrome C oxidase were increased. The release of ROS was decreased. The expression of Bcl-2 up-regulated and bax down-regulated. Conclusion: Weichangshu could promoted the gastrointestinal motility, which might be related with the regulation of mitochondrial kinetic energy.
Abstract:
Objective:To optimize matrix prescription of Guyanxiao cataplasm. Method: Matrix prescription was optimized by orthogonal test with initial viscosity,sustained adhesive strength and general sensory as comprehensive evaluation index,the amount of NP700,CMC-Na, glycerol and titania were chosen as factors. Result: Optimal matrix prescription was:NP-700, CMC-Na, glycerol and titania of 3,0.6,16,0.8g,respectively. Conclusion: Guyanxiao catapasm with good appearance and adhesion could be prepared by optimized matrix formulation.
Objective:To optimize matrix prescription of Guyanxiao cataplasm. Method: Matrix prescription was optimized by orthogonal test with initial viscosity,sustained adhesive strength and general sensory as comprehensive evaluation index,the amount of NP700,CMC-Na, glycerol and titania were chosen as factors. Result: Optimal matrix prescription was:NP-700, CMC-Na, glycerol and titania of 3,0.6,16,0.8g,respectively. Conclusion: Guyanxiao catapasm with good appearance and adhesion could be prepared by optimized matrix formulation.
2013,19(8):46-50, DOI: 10.11653/syfj2013080046
Abstract:
Objective: To optimize formulation of ursolic acid self-microemulsion drug delivery system using central composite design/response surface methodology. Method: With ratio of surfactants and cosurfactants and oil content as independent variables,particle size and saturated drug doading were taken as dependent variables,optimum formulation was screened by SAS9.1.3 software,and verified. Result: Optimized formulation of ursolic acid self-microemulsion was oil phase(MCT)-emulsifier(HS15)-cosurfactants(alcohol) 12.5:62.5:25. Conclusion: This optimized formulation of ursolic acid self-microemulsion drug delivery system could be obtained exactly and conveniently by central composite design response surface methodology,and established model had a reliable predictability.
Objective: To optimize formulation of ursolic acid self-microemulsion drug delivery system using central composite design/response surface methodology. Method: With ratio of surfactants and cosurfactants and oil content as independent variables,particle size and saturated drug doading were taken as dependent variables,optimum formulation was screened by SAS9.1.3 software,and verified. Result: Optimized formulation of ursolic acid self-microemulsion was oil phase(MCT)-emulsifier(HS15)-cosurfactants(alcohol) 12.5:62.5:25. Conclusion: This optimized formulation of ursolic acid self-microemulsion drug delivery system could be obtained exactly and conveniently by central composite design response surface methodology,and established model had a reliable predictability.
2014,20(8):15-18, DOI: 10.13422/j.cnki.syfix.2014080015
Abstract:
Objective: To optimize ultrafine grinding process of Chuanxiong Rhizoma. Method: Taking feeding amount, moisture content and grinding time as independent variables, average particle size as dependent variable, Box-Behnken response surface methodology was adopted to optimizeultrafine grinding process of Chuanxiong Rhizoma, its powder properties and hygroscopicity before and after smashed were investigated. Result: Optimum ultrafine grinding process was as follows:feeding amount1.3 kg, moisture content 4%, grinding time 19 min;Deviation between the predicted value(18.357 7 μm) and the measured value(19.187 μm) of average particle size was only 4.5%.Response angles of ordinary and ultrafine powder were (49.4±0.03) and (50.2±0.02) °, bulk densities were (0.429±0.003) and (0.363±0.005) g·mL-1, tap densities were (0.609±0.05) amd (0.556±0.10) g·mL-1, moisture equilibrium times were about 80 h. Conclusion: Optimized ultrafine grinding process was reasonable and feasible, preparation technology of ultrafine powder of Chuanxiong Rhizoma should add flow aid and other auxiliary materials in order to improve fluidity of powder, ultrafine grinding process should strictly control humidity.
Objective: To optimize ultrafine grinding process of Chuanxiong Rhizoma. Method: Taking feeding amount, moisture content and grinding time as independent variables, average particle size as dependent variable, Box-Behnken response surface methodology was adopted to optimizeultrafine grinding process of Chuanxiong Rhizoma, its powder properties and hygroscopicity before and after smashed were investigated. Result: Optimum ultrafine grinding process was as follows:feeding amount1.3 kg, moisture content 4%, grinding time 19 min;Deviation between the predicted value(18.357 7 μm) and the measured value(19.187 μm) of average particle size was only 4.5%.Response angles of ordinary and ultrafine powder were (49.4±0.03) and (50.2±0.02) °, bulk densities were (0.429±0.003) and (0.363±0.005) g·mL-1, tap densities were (0.609±0.05) amd (0.556±0.10) g·mL-1, moisture equilibrium times were about 80 h. Conclusion: Optimized ultrafine grinding process was reasonable and feasible, preparation technology of ultrafine powder of Chuanxiong Rhizoma should add flow aid and other auxiliary materials in order to improve fluidity of powder, ultrafine grinding process should strictly control humidity.
ZHAO Guozhen, LI Huizhen, LIANG Ning, ZHANG Haili, LIU Bin, CHE Qianzi, ZHOU Feng, LI He, CHEN Xiaowen, YE Long, LIN Jiahao, ZONG Xingyu, WANG Dingyi, SHI Nannan, WANG Yanping
2024,30(22):87-93, DOI: 10.13422/j.cnki.syfjx.20240824
Abstract:
The clinical practice guidelines of traditional Chinese medicine （TCM） have problems such as limited clinical application and unclear implementation effects， which may be related to the lack of clinical practice evidence. To provide reliable and precise evidence for clinical practice， this article proposes a model of combining TCM guidelines with real-world study， which includes 4 steps. Firstly， during the implementation process of the guidelines， a high-quality research database is established. Secondly， the recommendations in the guidelines are evaluated based on the established database in multiple dimensions， including applicability， effectiveness， safety， and cost-effectiveness， and thus their effectiveness in practical applications can be determined. Thirdly， based on the established database， core prescriptions are identified， and the targeted populations and medication plans are determined. That is， the best treatment regimen is established based on the analysis of abundant clinical data regarding the effects of different medication frequencies， dosages， and duration on efficacy. Fourthly， the guidelines are updated according to the real-world evidence. The research based on this model can provide real-world evidence for ancient and empirical prescriptions， improving their application in clinical practice. Moreover， this model can reduce research costs and improve research efficiency. When applying this model， researchers need to pay attention to the quality of real-world evidence， ensuring that it can truly reflect the situation in clinical practice. In addition， importance should be attached to the clinical application of guideline recommendations， ensuring that doctors can conduct standardized diagnosis and treatment according to the guidelines. Finally， full-process participation of multidisciplinary experts is encouraged to ensure the comprehensiveness and scientificity of the study. In conclusion， the application of this model will contribute to the development of TCM guidelines responsive to the needs of clinical practice and achieve the goal of promoting the homogenization of TCM clinical diagnosis and treatment.
The clinical practice guidelines of traditional Chinese medicine （TCM） have problems such as limited clinical application and unclear implementation effects， which may be related to the lack of clinical practice evidence. To provide reliable and precise evidence for clinical practice， this article proposes a model of combining TCM guidelines with real-world study， which includes 4 steps. Firstly， during the implementation process of the guidelines， a high-quality research database is established. Secondly， the recommendations in the guidelines are evaluated based on the established database in multiple dimensions， including applicability， effectiveness， safety， and cost-effectiveness， and thus their effectiveness in practical applications can be determined. Thirdly， based on the established database， core prescriptions are identified， and the targeted populations and medication plans are determined. That is， the best treatment regimen is established based on the analysis of abundant clinical data regarding the effects of different medication frequencies， dosages， and duration on efficacy. Fourthly， the guidelines are updated according to the real-world evidence. The research based on this model can provide real-world evidence for ancient and empirical prescriptions， improving their application in clinical practice. Moreover， this model can reduce research costs and improve research efficiency. When applying this model， researchers need to pay attention to the quality of real-world evidence， ensuring that it can truly reflect the situation in clinical practice. In addition， importance should be attached to the clinical application of guideline recommendations， ensuring that doctors can conduct standardized diagnosis and treatment according to the guidelines. Finally， full-process participation of multidisciplinary experts is encouraged to ensure the comprehensiveness and scientificity of the study. In conclusion， the application of this model will contribute to the development of TCM guidelines responsive to the needs of clinical practice and achieve the goal of promoting the homogenization of TCM clinical diagnosis and treatment.
Abstract:
Objective:To optimize matrix formulation of Sanhuang Sanyu cataplasm. Method: Taking appearance and adhesion of cataplasm as evaluation indexes,matrix formulation of Sanhuang Sanyu cataplasm was optimized by single factor test and orthogonal test. Result: Optimized matrix formulation of Sanhuang Sanyu cataplasm was NP-700-K90-glycerol(0.5∶0.4∶6).0.02 g·mL-1 solution was received by adding carbomer-u10NF into mixed extraction concentrated liquid of Rheum palmatum and Scutellaria baicalensis,swelling overnight, as A phase,0.5 g sodium polyacrylate and 0.4 g PVP were added to 6 g glycerol as B phase,aluminum chloride and citric acid were added to water as C phase,single extraction concentrated liquid of Phellodendron chinense was as D phase,four phases was fully mixed with A-B-C-D(10∶3∶3∶5), stirred 15 min, coating,covered with anti-sticking layer. Conclusion: Optimized matrix formulation of Sanhuang Sanyu cataplasm was simple,stable and feasible,which could be applied to industrial production.
Objective:To optimize matrix formulation of Sanhuang Sanyu cataplasm. Method: Taking appearance and adhesion of cataplasm as evaluation indexes,matrix formulation of Sanhuang Sanyu cataplasm was optimized by single factor test and orthogonal test. Result: Optimized matrix formulation of Sanhuang Sanyu cataplasm was NP-700-K90-glycerol(0.5∶0.4∶6).0.02 g·mL-1 solution was received by adding carbomer-u10NF into mixed extraction concentrated liquid of Rheum palmatum and Scutellaria baicalensis,swelling overnight, as A phase,0.5 g sodium polyacrylate and 0.4 g PVP were added to 6 g glycerol as B phase,aluminum chloride and citric acid were added to water as C phase,single extraction concentrated liquid of Phellodendron chinense was as D phase,four phases was fully mixed with A-B-C-D(10∶3∶3∶5), stirred 15 min, coating,covered with anti-sticking layer. Conclusion: Optimized matrix formulation of Sanhuang Sanyu cataplasm was simple,stable and feasible,which could be applied to industrial production.
Abstract:
Fracture healing is a process that involves the proliferation and differentiation of cells and tissues and can be stimulated by drugs. Traditional medicine has a reputation in the therapy on bone fracture. It is believed that herbal formulae promotes bone repair through multiple functions of “anti-inflammation”, “promoting blood circulation” and “stimulating bone formation”. This study aims to engineer a herbal formula under the guidance of Chinese medicine theories, then define the effective dosage ratio between the herbs and demonstrate the biological effects of the formulae on different parameters of healing using relevant in vitro assays. Six herbs which are frequently used for fractures were combined into five formulae according to the “Uniform Design” method. Assays of nitric oxide (NO) inhibition in mouse macrophages, human umbilical vein endothelial cells (HUVEC) and rat osteoblasts proliferation were used to reflect the “anti-inflammation inflammatory”, “circulation promotion” and “bone formation stimulation” functions. The results showed that all the formulae expressed significant activities of NO inhibition, and HUVEC cell and osteoblasts proliferation. Formula 5 with the largest doses (containing 30g Radix et Rhizoma Rhei, 9g Fructus Gardeniae, 9g Radix et Rhizoma Notoginseng, 9g Flos Carthami, 30g Ramulus Sambucus Willamsii, 15g Radix Dipsaci) demonstrated the best results in all the three assays.
Fracture healing is a process that involves the proliferation and differentiation of cells and tissues and can be stimulated by drugs. Traditional medicine has a reputation in the therapy on bone fracture. It is believed that herbal formulae promotes bone repair through multiple functions of “anti-inflammation”, “promoting blood circulation” and “stimulating bone formation”. This study aims to engineer a herbal formula under the guidance of Chinese medicine theories, then define the effective dosage ratio between the herbs and demonstrate the biological effects of the formulae on different parameters of healing using relevant in vitro assays. Six herbs which are frequently used for fractures were combined into five formulae according to the “Uniform Design” method. Assays of nitric oxide (NO) inhibition in mouse macrophages, human umbilical vein endothelial cells (HUVEC) and rat osteoblasts proliferation were used to reflect the “anti-inflammation inflammatory”, “circulation promotion” and “bone formation stimulation” functions. The results showed that all the formulae expressed significant activities of NO inhibition, and HUVEC cell and osteoblasts proliferation. Formula 5 with the largest doses (containing 30g Radix et Rhizoma Rhei, 9g Fructus Gardeniae, 9g Radix et Rhizoma Notoginseng, 9g Flos Carthami, 30g Ramulus Sambucus Willamsii, 15g Radix Dipsaci) demonstrated the best results in all the three assays.
Abstract:
Abstract:Objective: to investigate the absorption mechanism of palmatine chloride by using Caco-2 monolayer mode1.Methods: Caco-2 cell monolayer model was used to investigate the bi-directional transport of palmatine chloride. The effect of time, drug concentration, temperature, pH and inhibitors on the absorption of palmatine chloride was studied. Drug concentration was measured by HPLC and the apparent permeability coefficients (Papp) were calculated. Results: The apical to basolateral permeability (Papp) of palmatine chloride was bigger than the basolateral to apical permeability(Papp).Transport of palmatine chloride was time and concentration dependent, and it was also effected by P-glycoprotein inhibitor, pH and temperature. Conclusion:The absorption of palmatine chloride in Caco-2 cell model was an active transportation mediated by transporter, which mainly located in the apical side of Caco-2 cell monolayer.
Abstract:Objective: to investigate the absorption mechanism of palmatine chloride by using Caco-2 monolayer mode1.Methods: Caco-2 cell monolayer model was used to investigate the bi-directional transport of palmatine chloride. The effect of time, drug concentration, temperature, pH and inhibitors on the absorption of palmatine chloride was studied. Drug concentration was measured by HPLC and the apparent permeability coefficients (Papp) were calculated. Results: The apical to basolateral permeability (Papp) of palmatine chloride was bigger than the basolateral to apical permeability(Papp).Transport of palmatine chloride was time and concentration dependent, and it was also effected by P-glycoprotein inhibitor, pH and temperature. Conclusion:The absorption of palmatine chloride in Caco-2 cell model was an active transportation mediated by transporter, which mainly located in the apical side of Caco-2 cell monolayer.
Abstract:
Abstract：Objective To observe the effect of Dendrobium with different doses on ALT, AST, TC in the serum of chronic alcoholic liver injury mice. Methods Male ICR mice were divided into nine groups randomly: normal control group, model control group, positive control group, fresh Dendrobium groups with three doses(3g/kg,6g/kg,9g/kg), dried Dendrobium groups with three doses(0.45g/kg,0.90g/kg,1.35g/kg), gastric infusion for 65 days. The mice mode1 of chronic alcoholic 1iver injury was established by filling the stomach with alcohol after the administration for 65 days, once a day. The blood was taken to separate into serum and the activity of ALT, AST and the content of TC in the serum were determined. Results Compared with the normal control group, the ALT, AST, TC in the serum of model control group increased. Compared with model control group, fresh Dendrobium (3g/kg) and dried Dendrobium (0.45g/kg) could decrease ALT in the serum；fresh Dendrobium (3g/kg, 6g/kg) and dried Dendrobium (0.45g/kg) could decrease AST, TC in the serum; dried Dendrobium (0.90g/kg, 1.35g/kg) could decrease AST in the serum. Conclusion Fresh and dried Dendrobium could improve effectively the activity of ALT, AST and decrease the content of TC in the serum of chronic alcoholic 1iver injury mice.
Abstract：Objective To observe the effect of Dendrobium with different doses on ALT, AST, TC in the serum of chronic alcoholic liver injury mice. Methods Male ICR mice were divided into nine groups randomly: normal control group, model control group, positive control group, fresh Dendrobium groups with three doses(3g/kg,6g/kg,9g/kg), dried Dendrobium groups with three doses(0.45g/kg,0.90g/kg,1.35g/kg), gastric infusion for 65 days. The mice mode1 of chronic alcoholic 1iver injury was established by filling the stomach with alcohol after the administration for 65 days, once a day. The blood was taken to separate into serum and the activity of ALT, AST and the content of TC in the serum were determined. Results Compared with the normal control group, the ALT, AST, TC in the serum of model control group increased. Compared with model control group, fresh Dendrobium (3g/kg) and dried Dendrobium (0.45g/kg) could decrease ALT in the serum；fresh Dendrobium (3g/kg, 6g/kg) and dried Dendrobium (0.45g/kg) could decrease AST, TC in the serum; dried Dendrobium (0.90g/kg, 1.35g/kg) could decrease AST in the serum. Conclusion Fresh and dried Dendrobium could improve effectively the activity of ALT, AST and decrease the content of TC in the serum of chronic alcoholic 1iver injury mice.
Abstract:
Objective To establish a sensitive and specific HPLC method for quality control of Ilex hainanensis Merr.．Methods HPLC method was applied for quality assessment of Ilex hainanensis Merr.． HPLC analysis was performed on a HanbonC18 column (250mm×4．6mm，5 m)．The mobile phase consisted of acetonitrile (solvent B) and watercontaining0.1% (v/v) phosphoric acid (solvent A) at a constant flow rate of 1.0ml/min．An increasing linear gradient (v/v) of solvent B was used. The injection volume was 10μl. The column temperature was set at 25℃. The chromatogram s were monitored at 330nm. Results In different varieties of processing methods of Ilex hainanensis Merr. showing 9 characteristic peaks. It was established from its prepared samples of Ilex hainanensis Merr.．Conclusion The characteristic chromatogram of Ilex hainanensis Merr. with high specificity can be used to control its quality as well as the consistency of processing products.
Objective To establish a sensitive and specific HPLC method for quality control of Ilex hainanensis Merr.．Methods HPLC method was applied for quality assessment of Ilex hainanensis Merr.． HPLC analysis was performed on a HanbonC18 column (250mm×4．6mm，5 m)．The mobile phase consisted of acetonitrile (solvent B) and watercontaining0.1% (v/v) phosphoric acid (solvent A) at a constant flow rate of 1.0ml/min．An increasing linear gradient (v/v) of solvent B was used. The injection volume was 10μl. The column temperature was set at 25℃. The chromatogram s were monitored at 330nm. Results In different varieties of processing methods of Ilex hainanensis Merr. showing 9 characteristic peaks. It was established from its prepared samples of Ilex hainanensis Merr.．Conclusion The characteristic chromatogram of Ilex hainanensis Merr. with high specificity can be used to control its quality as well as the consistency of processing products.
Abstract:
Objective: To establish HPLC fingerprint of Tibetan medicine Pterocephali Herba, and compare it of Pterocephali Herba from different place, provided a new methods for its quality evaluation. Method: Adoption the method of HPLC to determine 10 groups of Pterocephali Herba from different place. Chromatographic Column was Boston Symmetrix(250×4.6mm,5μm), mobile phase was Methanol-0.1 mol.L-1 ammonium acetate solution(81:19),velocity was 0.8mL.min-1, the detection wavelength was 210nm, column temperature was 30℃. And compared the similarity of Pterocephali Herba from different place. Result: Taking advantage of < USP fingerprint similarity evaluation system (2004A version)> to establish common mode of HPLC fingerprint of Tibetan medicine Pterocephali Herba, and marked 101 characteristic peaks. The 10 groups samples had high similarity. Conclusion: This method had good repeatability, precision, and stability, providing scientific basis for quality control of Pterocephali Herba.
Objective: To establish HPLC fingerprint of Tibetan medicine Pterocephali Herba, and compare it of Pterocephali Herba from different place, provided a new methods for its quality evaluation. Method: Adoption the method of HPLC to determine 10 groups of Pterocephali Herba from different place. Chromatographic Column was Boston Symmetrix(250×4.6mm,5μm), mobile phase was Methanol-0.1 mol.L-1 ammonium acetate solution(81:19),velocity was 0.8mL.min-1, the detection wavelength was 210nm, column temperature was 30℃. And compared the similarity of Pterocephali Herba from different place. Result: Taking advantage of < USP fingerprint similarity evaluation system (2004A version)> to establish common mode of HPLC fingerprint of Tibetan medicine Pterocephali Herba, and marked 101 characteristic peaks. The 10 groups samples had high similarity. Conclusion: This method had good repeatability, precision, and stability, providing scientific basis for quality control of Pterocephali Herba.
Abstract:
Objective: To establish TLC fingerprint of antibacterial extraction of Pilea aquarum Dunn. Methods: The antibacterial activity extraction of Pilea aquarum Dunn was confirmed by antibacterial activity research, and then the TLC fingerprint of 10 batches of Pilea aquarum Dunn from Guizhou was analyzed comparatively. Results: The ethyl acetate extraction of Pilea aquarum Dunn is antibacterial activity. Another, the TLC chromatogram of 10 groups of drugs were very similar, which had 8 spots and 8 corresponding peaks, but the intensity of each peak was different. Conclusion: The method is simple and stabilized, and can be used for the quality control of Pilea aquarum Dunn.
Objective: To establish TLC fingerprint of antibacterial extraction of Pilea aquarum Dunn. Methods: The antibacterial activity extraction of Pilea aquarum Dunn was confirmed by antibacterial activity research, and then the TLC fingerprint of 10 batches of Pilea aquarum Dunn from Guizhou was analyzed comparatively. Results: The ethyl acetate extraction of Pilea aquarum Dunn is antibacterial activity. Another, the TLC chromatogram of 10 groups of drugs were very similar, which had 8 spots and 8 corresponding peaks, but the intensity of each peak was different. Conclusion: The method is simple and stabilized, and can be used for the quality control of Pilea aquarum Dunn.
Abstract:
Objecive To discuss the relationship between water-soluble sugar and cold-heat properties and furtherly reveal correlations of four properties and material basis.Methods Ten cold herbs and heat herbs were selected for the determination,and dealed with the water-soluble sugar with hydroxylamine hydrochloride,pyridine and acetic anhydride.Measuring Gas chomatography-mass(GC-MS) fingerprints of water-soluble sugar,then determining the water-soluble components of 20 herbs by mass spectrometry and analyzing the results by Fisher method.Results A discriminant function had been established by Fisher.The Fisher discriminant analysis could distinguish the cold and heat properties about 20 herbs that the group back substitution is 100%.From the results,it confirmes that there is difference between the cold and heat properties in the GC-MS fingerprints by Fisher method,which shows that there are different water-soluble sugar in the cold and the heat herbs.Conclusion The Fisher analysis could differentiate the cold and heat properties of herbs,and it is an effective statistical tool to study the relationship between the cold and heat properties and water-soluble sugar.It concludes that the water-soluble sugar is one of the material basis of four properties of herbs.
Objecive To discuss the relationship between water-soluble sugar and cold-heat properties and furtherly reveal correlations of four properties and material basis.Methods Ten cold herbs and heat herbs were selected for the determination,and dealed with the water-soluble sugar with hydroxylamine hydrochloride,pyridine and acetic anhydride.Measuring Gas chomatography-mass(GC-MS) fingerprints of water-soluble sugar,then determining the water-soluble components of 20 herbs by mass spectrometry and analyzing the results by Fisher method.Results A discriminant function had been established by Fisher.The Fisher discriminant analysis could distinguish the cold and heat properties about 20 herbs that the group back substitution is 100%.From the results,it confirmes that there is difference between the cold and heat properties in the GC-MS fingerprints by Fisher method,which shows that there are different water-soluble sugar in the cold and the heat herbs.Conclusion The Fisher analysis could differentiate the cold and heat properties of herbs,and it is an effective statistical tool to study the relationship between the cold and heat properties and water-soluble sugar.It concludes that the water-soluble sugar is one of the material basis of four properties of herbs.
Periodical information
ISSN：1005-9903
CN：11-3495/R
Editor in chief：
Proprieter：Tong Yan
Impact factor：
Publish house：Chinese Journal of Experimental Traditional Medical Formulae
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