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目的 研究小鼠暴发性肝衰竭 (FHF)动物模型凋亡肝细胞中凋亡基因胱门蛋白酶 (cas pase 3 )mRNA及蛋白的表达及意义。方法 尾静脉注射肿瘤坏死因子 α(TNF α)于D 氨基半乳糖(GalN)致敏的BALB/c小鼠 ,造成暴发性肝衰竭模型 ,用脱氧核糖核酸转移酶介导的缺口原位末端标记 (ISEL)技术、电镜及琼脂糖凝胶电泳检测肝组织DNALadder观察肝细胞凋亡 ,分别用免疫组化和半定量逆转录 聚合酶链反应 (RT PCR)方法检测凋亡基因caspase 3蛋白及mRNA表达情况。 结果 肝细胞凋亡率分别为 0 .0 %( 1.5h) ,0 .3 %( 3 .5h) ,3 .15 %( 6h)和 3 .19%( 9h) ,肝组织中caspase 3mRNA和蛋白表达与肝细胞凋亡率成正相关趋势。结论 在TNF α介导的小鼠暴发性肝衰竭模型中 ,caspase 3蛋白和mRNA表达上调并激活肝细胞凋亡的级联过程 ,最终导致肝细胞凋亡和坏死。
Objective To study the expression and significance of caspase 3 mRNA and protein in apoptotic hepatocytes induced by fulminant hepatic failure (FHF) in mice. Methods Tumor necrosis factor α (TNFα) was injected into the BALB / c mice sensitized with D-galactosamine (GalN) through the caudal vein, resulting in a model of fulminant hepatic failure using nick-end-transfected enzyme-mediated nick end labeling (ISEL), electron microscopy and agarose gel electrophoresis were used to detect the apoptosis of hepatocytes in DNALadder. The expressions of caspase 3 and mRNA were detected by immunohistochemistry and semi-quantitative reverse transcription polymerase chain reaction (RT PCR) Express the situation. Results The rates of hepatocyte apoptosis were 0 .0% (1.5h), 0.3% (3.5h), 3.15% (6h) and 3.19% (9h), respectively. The expressions of caspase 3 mRNA and protein Expression and apoptosis rate of hepatocytes have a positive correlation. Conclusions In the TNFα-mediated murine fulminant hepatic failure model, the caspase 3 protein and mRNA are up-regulated and a cascade of hepatocyte apoptosis is activated, eventually leading to hepatocellular apoptosis and necrosis.