熊去氧胆酸对急性肝功能衰竭大鼠肝细胞凋亡内质网途径的阻断作用

Restraining effects of ursodesoxycholic acid on endoplasmic reticulum stress route of hepatocyte apoptosis in rats with acute liver failure

目的探讨熊去氧胆酸对急性肝功能衰竭大鼠肝细胞凋亡内质网途径的阻断作用。方法以D-氨基半乳糖（D-Gal）腹腔注射一次性诱导大鼠出现急性肝功能衰竭模型,将模型鼠（60只）随机分为A、B两组,每组30只。A组为熊去氧胆酸药物治疗组,B组为空白对照组,于不同时间点检测大鼠的生存率和肝功能,采用琼脂糖凝胶电泳检测肝细胞DNA凋亡带,用半定量逆转录聚合酶链反应检测肝组织中Caspase-12 mRNA的表达水平。结果 D-Gal成功完成了大鼠急性肝功能衰竭模型,两组大鼠在24小时内无死亡,24-48小时死亡最多,96小时后两组均无死亡。B组24小时肝组织DNA梯度分析显示出现典型凋亡带,48小时时未减弱,72小时时开始减弱,直至7天后消失,ALT于48小时达到峰值,TBil水平变化不显著,ALB水平无变化,Caspase-12 mRNA在48小时亦达到峰值,随后其表达量逐渐降低。A组24小时与48小时时肝组织DNA梯度分析均显示非典型凋亡带,72小时后无显著凋亡带出现。ALT水平和Caspase-12 mRNA表达量与B组相比,差异均有统计学意义（P均〈0.05）。结论熊去氧胆酸对急性肝衰竭大鼠肝细胞凋亡内质网途径有明显阻断作用。

Objective To explore the restraining effect of ursodeoxycholic acid（UDCA） on endoplasmic reticulum stress route of hepatocyte apoptosis in rats with acute liver failure. Methods Acute liver failure rat model was induced by intraperitoneal injection of D-Gal. Total of 60 model rats were randomly divided into two groups（group A and group B）, 30 rats in each group. Group A was the ursodesoxycholic acid group and group B was blank control group. The survival rate and liver function of rats were detected at different time points. Agarose gel electrophoresis was used to observe hepatocyte DNA apoptosis band and semi-quantitative reverse transcription polymerase chain reaction was used to detect the expression level of Caspase-12 mRNA in liver tissues. Results Acute liver failure rat model was successfully induced by D-Gal. No death occurred in both groups within 24 h, most deaths occurred in 24-48 h, and no death occurred after 96 h. In the DNA gradient analysis for liver tissues of group B, typical apoptosis band appeared at 24 h, which did not attenuated at 48 h, began to attenuate at 72 h and disappeared after 7 d. ALT reached a peak at 48 h, with no significant changes in TBil and no changes in ALB. The expression of Caspase-12 mRNA also reached a peak at 48 h, and gradually decreased later. In the DNA gradient analysis for liver tissues of group A, atypical apoptosis band appeared at 24 h, the apoptosis band at 48 h was similar to that at 24 h, and no obvious apoptosis band was present after 72 h. Compared with group B, the differences of ALT and Caspase-12 mRNA levels were statistically significant（P 0.05）. Conclusion UDCA has obvious restraining effect on the endoplasmic reticulum stress route of hepatocyte apoptosis in rats with acute hepatic failure.