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目的 :研究肿瘤坏死因子α(TNF α)在暴发性肝衰竭肝细胞凋亡中的作用。方法 :对昆明小鼠注射脂多糖 (LPS)和D 氨基半乳糖 (D GalN) ,构建暴发性肝衰竭动物模型 ,采用脱氧核糖核酸转移酶介导的缺口原位末端标记技术及肝细胞DNA提取法 ,对肝细胞凋亡进行观察 ,并用RT PCR法检测肝组织TNF αmRNA的含量 ,用ELISA法测定血清中TNF α水平。结果 :在暴发性肝衰竭小鼠中 ,用药后 2h肝组织TNF αmRNA的表达达高峰 ,伴血清TNF α水平升高 ,以后逐渐下降 ,8h时以肝细胞凋亡为主 ,12h时则以肝细胞坏死为主 ,同时亦有肝细胞凋亡存在 ,应用抗TNF α抗体可阻断LPS +D GalN介导的肝细胞凋亡。结论 :在由LPS +D GalN所致的暴发性肝衰竭中肝细胞凋亡与TNF α的产生密切相关。肝细胞凋亡主要发生在病程早期 ,后期以坏死为主 ,凋亡可能是肝细胞坏死的早期过程
Objective: To study the role of tumor necrosis factor α (TNF α) in hepatocyte apoptosis in fulminant hepatic failure. Methods: Kunming mice were injected with lipopolysaccharide (LPS) and D-galactosamine (D GalN) to establish an animal model of fulminant hepatic failure. DNA nick end labeling (TdT) Method, the apoptosis of hepatocytes was observed. The content of TNFαmRNA in liver tissue was detected by RT-PCR, and the level of TNFα in serum was determined by ELISA. Results: In the fulminant hepatic failure mice, the expression of TNFαmRNA reached a peak at 2h after treatment, and the level of serum TNFα increased with the increase of serum TNFα level. The level of TNFαmRNA gradually decreased at 8h, The main cell necrosis, but also the presence of hepatocyte apoptosis, anti-TNF α antibody can block LPS + D GalN-mediated hepatocyte apoptosis. Conclusions: Hepatocyte apoptosis is closely related to the production of TNFα in fulminant hepatic failure induced by LPS + D GalN. Apoptosis of hepatocytes mainly occurs in the early course of disease, necrosis in the later period, apoptosis may be the early course of hepatocyte necrosis