二至丸预防和治疗性给药对大鼠损伤后肝细胞再生障碍的保护作用

目的:观察二至丸预防和治疗性给药对大鼠损伤后肝细胞再生障碍的保护作用,并探讨其作用机制。方法:将50只Wistar大鼠随机为5组,分别为正常组,模型组,二至丸预防组(6.48 g·kg~(-1)),二至丸治疗组(6.48 g·kg~(-1)),雷帕霉素组(1 mg·kg~(-1))。除正常组建立肝部分切除术(partial hepatectomy,PHx)模型外,其余各组ig给予2-乙酰氨基芴(2-acetylaminofluorene,2AAF)ig 7 d,同时二至丸预防组预防性给药7 d,第8天行肝部分切除术建立损伤后肝细胞再生抑制复合模型(2AAF/PHx),预防组和治疗组分别于术后24 h再行预防性给药组及治疗性给药3 d。大鼠处死后计算肝重指数,观察肝脏病理形态学变化,收集大鼠血清并采用自动生化分析仪检测血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST)及白蛋白。同时采用流式检测肝细胞凋亡水平及天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)表达。结果:与正常组比较,模型组大鼠肝重指数和白蛋白水平明显降低,ALT和AST水平明显升高,大鼠肝细胞坏死细胞(non-viable non-apoptotic cell,NVNAC),早期凋亡细胞(viable apoptotic cell,VAC),晚期凋亡细胞(non-viable apoptotic cell,NVAC)水平及Caspase-3表达明显降低,而活性细胞(living cell,LC)则明显降低(P0.05,P0.01),肝脏病理损伤较为明显;与模型组比较,二至丸预防组及治疗组均可明显升高大鼠肝重指数和白蛋白水平,同时降低其ALT和AST水平,且大鼠NVNAC,VAC,NVA水平及Caspase-3表达明显降低(P0.05,P0.01),而LC则明显升高(P0.05,P0.01),病理学观察提示明显好转。结论:二至丸预防性及治疗性给药可显著抑制损伤后肝细胞再生障碍大鼠肝细胞凋亡,并降低Caspase-3表达,肝功能损伤得以恢复,提示二至丸对大鼠损伤后肝细胞再生障碍具有良好的预防保护作用。

Protective Effect of Erzhiwan by Two Different Administrations for Rats with Liver Cytothesis Obstacle After Injury

Objective: To observe the protective effects of Erzhiwan by prophylactic and therapeutic administrations for the liver cytothesis obstacle in rats after injury, and investigate its mechanism. Method: The 50 Wistar rats were randomly divided into 5 groups:normal group, model group, Erzhiwan prophylactic group (6.48 g·kg^-1), Erzhiwan therapeutic group (6.48 g·kg^-1), and Rapamycin group (1 mg·kg^-1). In normal group, partial hepatectomy (PHx) was conducted to establish PHx models; in the other groups, animals received intragastric administration of 2-acetylaminofluorene/partial hepatectomy (2-AAF) for 7 days; meanwhile, in the Erzhiwan prophylactic group, those rats were administrated with Erzhiwan for 7 days, and on the 8th day, PHx was conducted to establish compound models of liver cytothesis obstacle after injury (2AAF/PHx models). 24 h after surgery, the rats in the prophylactic and therapeutic groups were administrated with Erzhiwan for three days. After rats were killed, the index of liver weight was calculated; liver pathomorphological changes were observed; and the levels of alanine transarninase (ALT), aspartate aminotransferase (AST) and albumin in serum were detected by using automatic biochemical analyzer. In addition, liver apoptosis and Caspase-3 expression levels were analyzed by flow cytometry. Result: As compared with the normal group, the index of liver weight, albumin level and count of living cell(LC) were significantly decreased, while ALT and AST levels, non-viable non-apoptotic cell(NVNAC),viable apoptotic cell(VAC),non-viable apoptotic cell(NVAC) and Caspase-3 expression levels were significantly incereased in model group (P <0.05,P <0.01). As compared with the model group, the index of liver weight, level of albumin and count of LC were increased, however, the levels of ALT, AST, NVNAC,VAC,NVAC and Caspase-3 expression levels were decreased in both Erzgu pill prophylactic and therapeutic groups (P <0.05,P <0.01). Meanwhile pathological observation hinted that injured livers were improved markedly. Conclusion: Erzhiwan remarkably inhibited liver apoptosis, and decreased Caspse-3 expression of 2-AAF/PHx rats to remit damaged liver function. So Erzhiwan play a favorable and protective role in treatment of cytothesis obstacle of damaged liver in rats