生地黄药汁炮制降低雷公藤肝毒性的作用机制

Mechanism of Rehanniae Radix drug decoction on reducing the hepatotoxicity induced by Tripterygium wilfordii Hook.f.

目的:探讨生地黄药汁制法炮制对雷公藤诱导的肝毒性的减毒作用及其机制。方法:60只SPF级雄性ICR小鼠随机分为正常组、雷公藤生品组(2 g/kg)、生地黄药汁制雷公藤1、2倍剂量组(2、4 g/kg)、清水制雷公藤组(2 g/kg)和生地黄药汁辅料组(1 g/kg),各组均灌胃相应药物,1次/d,连续28 d,正常组给予等体积的溶媒,通过对小鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)及肝组织病理的检测分析,初步评价其减毒作用;进一步借助蛋白免疫印迹法及试剂盒方法,通过对肝凋亡相关蛋白及脂质过氧化和抗氧化指标的检测分析,评价其减毒机制。结果:与正常组比较,雷公藤生品组致使小鼠肝脏系数、血清ALT、AST水平均显著升高,出现肝细胞变性,并伴有炎性浸润及血管瘀血,显著上调肝B淋巴细胞瘤-2(Bcl-2)相关X蛋白(Bax)表达水平,下调肝Bcl-2蛋白表达水平,升高丙二醛(MDA)水平,降低超氧化物歧化酶(SOD)水平(P<0.01),而经1、2倍剂量生地黄药汁制雷公藤炮制品干预后均在不同程度上逆转了上述指标的异常(P<0.01,P<0.05)。结论:生地黄药汁制雷公藤可能通过抑制肝促凋亡蛋白、脂质过氧化水平,增强抗凋亡蛋白及抗氧化防御从而发挥保肝作用。

Objective:To investigate the attenuating effect of processing with Rehanniae Radix on the hepatotoxicity induced by Tripterygium wilfordii Hook.f.(TWH),and to explore its attenuation mechanism.Methods:Sixty SPF grade male ICR mice were randomly divided into the control group,the raw TWH(RTWH)group(2 g/kg),Rehanniae Radix decoction processed TWH group(2,4 g/kg),water processed TWH(WTWH)group(2 g/kg),and the auxiliary group in Rehanniae Radix decoction group(1 g/kg).Each group was administered the corresponding drugs by gavage,once a day for 28 consecutive days,and the control group was given an equal volume of solvent.The detoxification effect was evaluated by the detection and analysis of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and liver histopathology of mice in each group.Further,the detoxification mechanism was preliminarily evaluated by the detection and analysis of some related indicators of apoptosis and oxidation/antioxidant related indexes of liver by Western Blot and kit methods.Results:Compared with the control group,the RTWH group significantly increased the liver coefficient,serum ALT and AST levels of mice,caused hepatocyte degeneration,accompanied by inflammatory infiltration and vascular congestion.Also,it significantly up-regulated the hepatic Bcl-2 related X protein(Bax)expression protein level,and down-regulated the hepatic B-lymphoma-2(Bcl-2)expression protein level,elevated the level of malondialdehyde(MDA),and decreased superoxide dismutase(SOD)level(P<0.01).However,after the intervention of the products of Rehanniae Radix decoction processed TWH,the abnormalities of the above indexes were reversed to varying degrees(P<0.01,P<0.05).Conclusion:Rehanniae Radix drug decoction processed TWH exerts hepatoprotective effects by inhibiting the hepatic pro-apoptotic protein,hepatic lipid peroxidation,enhancing the anti-apoptotic protein and antioxidant defense.