截断逆挽方降低慢加急性肝衰竭大鼠死亡率的机制探讨

Truncated inverse pull side to reduce the mechanism of the slow increase of the mortality of acute liver failure in rats

目的:探讨截断逆挽方降低慢加急性肝衰竭大鼠24 h死亡率,增加生存时间的作用机制。方法:SPF级Wistar大鼠150只,分为正常组、模型组、中药组。人血清白蛋白建立大鼠肝硬化模型后,中药组给予截断逆挽方灌胃3 d。再给予中药组和模型组D-GalN 400 mg/Kg加LPS 100μg/Kg一次性腹腔联合注射。注药后模型组和中药组各分为4组:4、8、12 h后各麻醉后处死1组,取肝组织进行HE染色和原位细胞凋亡检测(TUNEL)技术,并计算肝细胞的凋亡指数(AI);余观察24 h死亡率并记录生存时间。结果:与正常组比较,模型组各时间点HE染色可见肝内假小叶广泛形成,再生结节内肝细胞坏死、炎性浸润及出血明显,12 h最为显著,表现为大块或亚大块坏死;TUNEL检测见大量的凋亡小体、凋亡细胞,AI均显著增高(P<0.01);与模型组比较,中药组各对应时间点HE染色可见坏死和出血明显减轻,炎性细胞浸润减少;TUNEL检测中药组4 h、8 h凋亡细胞减少,12h坏死面积缩小,AI均降低(P<0.05)。中药组较模型组24 h死亡率降低,生存时间延长。结论:截断逆挽方降低ACLF大鼠24 h死亡率,延长其存活时间的机制可能与阻断肝凋亡细胞过程,或延缓肝细胞凋亡向死亡转化有关。

Objective： To investigate the truncated inverse pull side to reduce the acute on chronic liver failure in rats 24 h,mortality,and increase the survival time of the mechanism of action.Method： Wistar rats of SPF grade 150,divided into normal group,model group,the chinese medicines board.Human serum albumin in rat liver cirrhosis model,traditional chinese medicine group was given the truncated inverse pull side gavage for 3 days.Given herbal group and model group D-GalN.400 mg/Kg plus LPS-100 μg/Kg intraperitoneal joint injection.After injection model group and the group were divided into 4 groups： executed a set of 4,8,12 h after each anesthesia of liver tissue HE staining and in situ detection of apoptosis（TUNEL） techniques,and calculation of the liver cells the apoptotic index（AI）;I observed 24 h mortality and record survival time.Results： Compared with normal group,model group at all time points HE staining showed intrahepatic pseudolobule widely formation of regenerative nodules within the liver cell necrosis,inflammatory infiltration and hemorrhage significantly,12 h,most significantly,the performance for large pieces or sub-chunk necrosis;TUNEL assay to see a large number of apoptotic bodies and apoptotic cells,the AI were significantly increased（P0.01）;compared with model group,HE staining of the TCM group the corresponding time point of necrosis and hemorrhage significantly reduced inflammatory cell infiltration reduced;TUNEL assay medicine group for 4 h,8 h,apoptotic cells decreased 12 h,the necrotic area was narrow,the AI decreased（P 0.05）.TCM group than in model group 24 h mortality,and prolonged survival.Conclusion： The truncated inverse pull side reduce the ACLF rats 24 h mortality,prolong the survival time of the mechanism may block the cellular processes of hepatic apoptosis,or delay the apoptosis of liver cell transformation related to the death.