暴发性肝功能衰竭小鼠模型的建立及其治疗研究

目的:建立暴发性肝功能衰竭(FLF)小鼠模型并探讨复方甘草酸苷(SNM C)的保护作用及可能的作用机制。方法:采用D-氨基半乳糖(D-G a ln)和脂多糖(LPS)一次性腹腔注射构建FLF小鼠模型。利用光镜、电镜观察小鼠肝损伤情况;应用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测肝细胞原位凋亡情况;应用免疫组化法分别检测肝组织中细胞色素C和天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)的表达。结果:D-G a ln加LPS可成功构建FLF小鼠模型。光镜及电镜可见肝细胞大量凋亡,且凋亡情况随治疗时间的延长明显改善。TUNEL检测结果表明,随着SNM C治疗时间的延长,凋亡指数逐渐降低。细胞色素C和caspase-3在模型组细胞阳性表达明显增加,随SNM C治疗时间延长表达逐渐减少。结论:利用D-G a ln和LPS可以构建理想的FLF小鼠模型。SNM C能有效抑制小鼠FLF模型中的肝细胞凋亡。SNM C可能通过稳定线粒体膜抑制细胞色素C释放及其随后caspase-3活化,从而阻断肝细胞凋亡的进行。

Establishment of reduplicated model of fulminant liver failure and research on its treatment

Objective: To reduplicate the models of fulminant liver failure(FLF) of mice and investigate the protective effect of stronger neo-minophagen C(SNMC) on FLF.Methods: D-galactosamine(D-Galn) and lipopolysaccharide(LPS) were injected into the abdominal cavity of mice once to establish experimental model of FLF.The damage of liver was detected by light microscope and electron microscope.The hepatocyte apoptosis in-situ was estimated by terminal deoxynucleotidyl-transferase mediated dUTP-biotin nick end(labeling)(TUNEL).Cytochrome C(CYC) and caspase-3 in liver tissues were determined by immuno(histochemistry.) Results: D-Galn and LPS could be used to reduplicate the models of FLF.In the model,a lot of hepatocyte apoptosis was estimated by light microscope and electron microscope,and improved with(prolongation) of the therapeutic time.TUNEL assay revealed that the apoptosis index was gradually decreased after the prolongation of the therapeutic time of SNMC.The cellular positive expressions of CYC and(caspase-3) were strong in the model group,and they were decreased with prolongations of SNMC therapeutic time in the treatment group.Conclusion: D-Galn and LPS can be used to establish a perfect model of FLF.SNMC can effectively inhibit the hepatocyte apoptosis in the models of FLF.Its inhibitions of the release of CYC and the activation of caspase-3 are possibly by stabilizing the membrane of mitochondria inhibiting(release) of CYC and activation of caspase-3,thus the progress of hepatocyte apoptosis can be prevented.