固有免疫信号因子TRIF在肝纤维化中的表达  

Expression of TRIF of innate immunity signal pathway in hepatic fibrosis

作　　者：黄欣[1] 李艳[1] 贺雄[1] 赵苹利[1] 曹文富[1] 

机构地区：[1]重庆医科大学中医药学院,重庆401331

出　　处：《中国实验动物学报》2012年第4期26-30,I0004,共6页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金预研项目(NSFYY201108)

摘　　要：目的初步探讨TOLL样受体下游重要信号因子TRIF与肝纤维化发生发展的病理机制关系。方法以四氯化碳皮下注射+低蛋白高脂饮食+酒精饮料的方法复制大鼠肝纤维化模型。将SD大鼠随机分成正常对照组和模型组,在完成制备模型实验后进行取材。部分实验鼠进行心脏生理盐水和多聚甲醛灌注后,取肝脏组织制备石蜡切片,进行常规HE染色和免疫组化实验;另一部分实验鼠脱颈安乐死后,取新鲜肝组织进行电镜标本的制备和Western Blot实验检测。结果 HE染色结果显示与而正常对照组大鼠相比,模型组大鼠肝小叶结构明显破坏,肝细胞数量明显减少和肝纤维化程度等特点明显;电镜结果也显示,肝纤维化组可见大量胶原纤维沉积现象,胞质可见明显的溶解现象,在狄氏腔内,肝星状细胞的细胞核溶解,血窦内皮细胞胞质、胞核皆溶解;免疫组化模型组大鼠肝组织均显示内皮细胞、星形细胞等TRIF都有强烈高表达,并且以胞核表达为主,亦见胞质表达,而正常组呈现弱阳性表达;与正常对照组相比,模型组大鼠肝组织TRIF蛋白表达都明显升高,呈显著性差异(P<0.01),与形态学的表达特点相一致。结论 TRIF在肝纤维化中表达显著增强,说明在肝纤维化过程中,TOLL样受体明显激活,并且通过下游信号转导途径,在机体内产生一系列的免疫应答反应。通过该现象的观察,我们初步证实了TOLL样受体固有免疫信号因子TRIF在纤维化形成中起着重要的作用。Objective To study the characteristics of expression of TRIF（TIR-domain-containing adapter-inducing interferon-β,TRIF） innate immune signaling factors in hepatic cells and its protein expression pattern in hepatic fibrosis in rats,and to explore the relationship between the important signal downstream of TOLL-like receptor TRIF and pathogenetic mechanism of hepatic fibrosis.Methods To generate rat models of hepatic fibrosis by subcutaneous injection of carbon tetrachloride together with low protein and high fat diet and alcohol water drinking.Thirty two SD rats were randomly divided into control group and model group.Intracardiac saline and formaldehyde fixative perfusion was performed in some rats and liver tissue samples were taken for histopathological and immunohistochemical examination.Other rats were killed by direct decapitation and fresh liver tissue samples were taken for electron microscopic examination and Western blot assay.Results The HE staining showed that normal rat liver displaying regular lobular structure without inflammatory cell infiltration,but the model rat livers showed distinct disordered lobular structure,decreased amount of hepatocytes and the presence of hepatic fobrosis.Electron microscopic observation revealed deposition of collagen fibers and cytolysis of hepatocytes,karyolysis of Kuppfer cells,cytolysis of sinusoidal endothelial cells and a large number of abnormal cells infiltration.Immunohistochemistry showed a high expression of TRIF in endothelial and stellate cells,mainly,in the cell nuclei,but also could be seen in the cytoplasm.Compared with the normal control group,the TRIF protein expression was significantly increased in the model rat livers（P〈0.01）,consistent with that observed by histopathology.Conclusions TRIF expression is significantly increased in liver fibrosis,indicating an apparent activation of Toll-like receptor and a series of immunological responses via the downstream signal transduction pathways,producing a series of in vivo immune responses.It is

关 键 词：肝纤维化 固有免疫 TOLL-LIKE RECEPTORS TRIF 

分 类 号：Q95-33[生物学—动物学] R243[医药卫生—中医临床基础]