苦参碱联合甘草甜素在四氯化碳慢性肝损伤中的保护作用及机制

Protective Effect and Mechanism of Matrine Combined with Glycyrrhizic Acid in the Treatment of Chronic Liver Injury Induced by Carbon Tetrachloride

目的:考察苦参碱联合甘草甜素在四氯化碳(CCl_4)慢性肝损伤中的保护作用,并从能量代谢及CYP酶的角度探讨其保护机制。方法:建立CCl_4慢性肝损伤模型,通过考察血清ALT、AST观察两药及其联合用药在慢性肝损伤模型中的保护作用;检测血清谷氨酸脱氢酶(GLDH)及肝组织中肝脏腺嘌呤核苷三磷酸(ATP)、二磷酸腺苷(ADP)、腺嘌呤核糖核苷酸(AMP)含量,评价药物对肝脏能量代谢及线粒体功能的调节作用;实时定量PCR及Western Blot法检测肝脏CYP1A2、CYP2E1 mRNA及蛋白水平,评价两药及其联合用药对肝脏CYP酶的调控作用。结果:苦参碱(72.8 mg·kg^(-1))、甘草甜素(43.4 mg·kg^(-1))在CCl_4慢性肝损伤模型中均可降低大鼠血清ALT、AST(P<0.05),两药联合(36.4 mg·kg^(-1)苦参碱+21.7 mg·kg^(-1)甘草甜素)使用保护作用更加显著(P<0.05);其中苦参碱(72.8 mg·kg^(-1))、甘草甜素(43.4 mg·kg^(-1))均可降低血清GLDH,并恢复肝脏ATP含量(P<0.05);苦参碱(72.8 mg·kg^(-1))对CYP1A2、CYP2E1 mRNA表达水平无抑制作用,甘草甜素(43.4mg·kg^(-1))对CYP1A2、CYP2E1 mRNA及蛋白表达水平均有抑制作用(P<0.05)。结论:苦参碱联合甘草甜素在慢性肝损伤模型中具有明显的线粒功能调节和肝保护作用。

Objective： To investigate the protective effects of matrine combined with glycyrrhizic acid on chronic liver injury induced by carbon tetrachloride, and explore the protective mechanism from the points of energy metabolism and CYP enzyme. Methotis： The chronic hepatic injury model of rats was induced by CCl4. The changes of activity of alanine aminotransferase （ALT） and as- partate aminotransferase （AST） in serum were measured to observe the protective effect of the two drugs and their combination. The contents of glutamate dehydrogenase （GLDH） in serum and adenine nucleoside three phosphate （ATP）, adenosine diphosphate （ADP） and adenine monophosphate （AMP） in liver tissue were determined to evaluate the regulation effect on hepatic energy metabolism and mitochondrial function. The levels of CYP1A2, CYP2E1 mRNA and protein in liver tissue were detected by real-time PCR and Western Blot to evaluate the two drugs and their combination on the regulation function of liver CYP enzyme. Results： Matrine （ 72.8 mg·kg^-1） and glycyrrhizic acid （43.4 mg·kg^-1） could decrease the serum activities of ALT and ALT in chronic hepatic injury model, and the combination （matrine 36.4 mg·kg^-1 ＋ glycyrrhizic acid 21.7 mg·kg^-1） had the most significant protective effect （P 〈 0.05 ）. Matrine （72.8 mg·kg^-1） and glycyrrhizic acid （43.4 mg·kg^-1 ） could decrease GLDH in serum, and restore the content of ATP in liver （ P 〈 0.05 ）. Matrine （72.8 mg·kg^-1 ） had no effect on the expression of CYP1 A2 and CYP2E1 mRNA, and gly- cyrrhizin （43.4 mg·kg^-1 ） could inhibit the expression of CYP1A2, CYP2E1 mRNA and protein （ P 〈 0.05 ）. Conclusion： Matrine combined with glycyrrhizin has obvious regulation effect on mitochondrial function and liver protective effect in chronic hepatic injury model.