附子与人参不同配伍对心肌细胞的减毒作用

Toxicity-reducing Effect of Compatibility of Aconiti Lateralis Radix Praeparata with Different Proportion of Panax Ginseng in Neonatal Rat Cardiomyocytes

摘要:

目的: 探讨附子配伍不同比例人参的减毒作用. 方法: 将原代培养6 d的心肌细胞分为空白(NC)组,不同比例的附子人参(AG)组,空白血清(BS)组.AG组加入含有不同配伍比例的附子人参的载药血清,观察药物对原代培养心肌细胞的自发搏动的影响,检测心肌细胞的细胞活力,测定原代心肌细胞中超氧化物歧化酶(SOD)活性与丙二醛(MDA)含量,测定上清液中一氧化氮(NO)分泌量和乳酸脱氢酶(LDH)释放率,利用分光光度法检测半胱氨酸蛋白酶(Caspases-3)活性,Western blot法检测Bcl-2和Bax的蛋白表达,RT-PCR检测Bcl-2和Caspases-3 mRNA的表达. 结果: 与附子-人参按1 :0配伍的载药血清(AG_{1 :0})组比较,加入附子配伍不同比例人参载药血清后,心肌细胞自发搏动频率显著降低,心肌细胞内SOD活性显著升高,MDA含量,NO分泌量和LDH释放率显著降低,Caspases-3活性,Bax蛋白表达和Caspases-3 mRNA表达显著降低,Bcl-2蛋白表达和Bcl-2 mRNA表达显著升高,细胞活力在作用2~4 h显著升高,均为显著性差异(P <0.05).AG_1:0.5组对心肌细胞以上各项检测指标的影响与BS组比较不具有显著性差异. 结论: 附子配伍不同比例的人参可抑制附子对心肌细胞的毒性作用,其中附子-人参配比为1:0.5时,人参可有效地抑制附子的毒性作用.

Abstract:

Objective: In order to evaluate the inducing toxicity effect of Aconiti Lateralis Radix Praeparata and different compatibility proportion of Ginseng Radix et Rhizoma in neonatal rats cardiomyocytes. Method: The primary culture cardiac cells (6 d) were randomly divided into normal(NC) group, Aconiti Lateralis Radix Praeparata and Ginsang Radix et Rhizoma(AG) group, blood serum(BS) group. The inducing toxicity effect of blood serum of different compatibility proportion of Aconiti Lateralis Radix Praeparata and Ginseng Radix et Rhizoma on spontaneous imp frequency, cell viability, superoxde dismutase(SOD), malondialdehyde(MDA), Nitric oxide (NO) and lactate dehydrogenase(LDH) was investigated in vitro in neonatal rat cardiomyocytes. The activity of Caspases-3, the expression of Bcl-2 and Bax protein, the expression of Bcl-2 and Caspases-3 mRNA was assayed by spectrophotometry, western blot and real-time PCR respectively. Result: Compared with AG_1:0 group, the content of SOD increased obviously, spontaneous imp frequency, the content of MDA, the secretory volume of NO and the release rate of LDH decreased obviously, the activity of Caspases-3, the expression of Bax protein and Caspases-3 mRNA decreased obviously, the expression of Bcl-2 protein and Bcl-2 mRNA increased gradually, cell viability increased in 2-4 h significantly (P <0.05) in other AG groups. Compared with BS group, there was no significant difference in AG_{1 : 0.5} group in the above indexes. Conclusion: There was a significant inducing toxicity effect of Aconiti Lateralis Radix Praeparata and different compatibility proportion of Ginseng Radix et Rhizoma in neonatal rats cardiomyocytes, the optimization compatibility proportion in AG_1:0.5.