冰片对交泰丸在抑郁模型大鼠药效学、主要活性成分药动学及脑组织分布的影响

Effect of Borneol on Pharmacodynamics，Pharmacokinetics and Brain Tissue Distribution of Main Active Ingredients of Jiaotaiwan in Depression Model Rats

摘要:

目的 探讨交泰丸与冰片联用时，对脂多糖（LPS）诱导抑郁模型大鼠药效学，以及7种主要成分药代动力学及脑组织分布的影响。方法 将40只SD大鼠随机分为空白组、模型组、氟西汀组（10 mg·kg^-1）、交泰丸组（1.50 g·kg^-1）、交泰丸冰片联用组（交泰丸1.50 g·kg^-1+冰片0.05 g·kg^-1），每组8只。除空白组外采用腹腔注射LPS建立抑郁模型，各组分别灌胃给予相应药物或等体积纯水，连续14 d。采用糖水偏好实验、旷场实验及强迫游泳实验评估大鼠的抑郁状态；采用酶联免疫吸附测定法（ELISA）检测大鼠血清中白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）水平；苏木素-伊红（HE）和尼氏（Nissl）染色观察大鼠海马组织神经元形态变化。取行为学实验后的交泰丸组和交泰丸冰片联用组，分别于灌胃后在设置的时间点采血，采用超高效液相色谱-三重四极杆串联质谱法（UPLC-QqQ-MS/MS）测定血浆中药根碱、表小檗碱、黄连碱、巴马汀、小檗碱、木兰花碱、小檗红碱的含量，采用DAS 3.2.2软件对各成分药代动力学参数进行分析。采血完成后冰上快速取脑，采用UPLC-QqQ-MS/MS比较7种成分在脑组织中的含量变化。结果 与空白组比较，模型组大鼠糖水偏好率降低、旷场总路程减少、游泳不动时间延长，血清中炎症因子表达升高（P<0.01）；与模型组比较，各给药组大鼠糖水偏好率增加、旷场总路程增加、游泳不动时间缩短，交泰丸组和交泰丸冰片联用组大鼠血清中炎症因子表达降低（P<0.05，P<0.01）。病理观察结果显示，交泰丸组和交泰丸冰片联用组均对海马神经元有一定保护作用。药动学结果显示，与交泰丸组比较，交泰丸冰片联用组中小檗碱、表小檗碱的药时曲线下面积（AUC_0-t、AUC_0-∞）、达峰浓度（C_max）、平均稳态血药浓度（C_av）增加（P<0.05，P<0.01），黄连碱的AUC_0-t、AUC_0-∞、平均驻留时间（MRT_0-∞）、C_av增加（P<0.05，P<0.01），药根碱的C_max明显升高（P<0.05）；但7种生物碱的药动学变化趋势一致。脑组织分布检测结果显示，与交泰丸组比较，交泰丸冰片联用组大鼠脑组织中药根碱、表小檗碱、黄连碱、巴马汀、小檗碱含量升高（P<0.05，P<0.01）；木兰花碱含量升高，小檗红碱含量下降，但差异无统计学意义。结论 交泰丸单用及与冰片联用均能改善大鼠抑郁样行为，减轻血清炎症因子水平，改善海马区病理损伤，具有抗抑郁作用；与冰片联用后，可增加交泰丸中多种有效成分在大鼠血浆及脑组织中的暴露量，提高其生物利用度，促进其吸收，增加药物在脑组织中的分布，更有利于交泰丸发挥抗抑郁作用。

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（AUC_0-t， AUC_0-∞）， peak concentration（C_max） and the average steady-state plasma concentration（C_av） of berberine and epiberberine in the combination group were increased（P<0.05， P<0.01）， AUC_0-t， AUC_0-∞， mean residence time（MRT_0-∞） and C_av of coptisine were significantly increased（P<0.05， P<0.01）， C_max 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.