实验性自身免疫性肝炎小鼠模型Fas/FasL的表达及凋亡研究  

The research of Fas /FasL expression and apoptosis in experimental autoimmune hepatitis mouse model

作　　者：艾国[1] 肖芳[1] 李兰[1] 陶然[1] 黄加权[1] 严伟明[1] 宁琴[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院感染科,感染性疾病研究所,湖北武汉430030

出　　处：《中西医结合肝病杂志》2013年第4期211-213,I0001,共4页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：卫生部临床学科重点项目[No.卫规财函(2010)439号];湖北省自然科学基金项目(No.2012FFB02319)

摘　　要：目的:建立实验性自身免疫性肝炎小鼠模型(Experimental autoimmune hepatitis,EAH),观察Fas/FasLigand(Fas L)的表达及其介导的细胞凋亡在自身免疫肝组织损伤中的作用。方法:以同种系肝抗原与弗氏完全佐剂充分乳化后两次经腹腔注入到C57BL/6小鼠体内建立EAH,观察其血清ALT、AST水平和肝组织病理学改变;免疫荧光检查血清中自身抗体;采用实时荧光定量-聚合酶链反应(Realtime-PCR)和免疫组化方法检测小鼠肝脏组织中Fas与FasL基因转录与蛋白的表达,TUNEL(脱氧核糖核酸末端转移酶介导的缺口末端标记)法检测小鼠肝组织细胞凋亡情况。结果:建模后第28天的模型组小鼠肝脏组织中,主要表达于肝细胞及浸润淋巴细胞的Fas和FasL在mR-NA及蛋白水平均明显增强(P<0.05);光镜观察见正常小鼠肝脏组织无异常改变,而模型组小鼠可见散在或局灶性的炎性细胞浸润,并于炎性区域中心及周围出现TUNEL阳性的肝细胞和淋巴细胞,模型组小鼠肝组织内细胞凋亡指数(Apoptosis Index,AI)明显升高。结论:实验性自身免疫性肝炎小鼠肝组织炎症浸润与细胞凋亡共存,通过Fas/FasL介导的细胞凋亡与自身免疫性肝炎发生发展有关。Objective： To establish experimental autoimmune hepatitis （EAH） mouse model and observe Fas/FasL expres- sion and cell apoptosis in autoimmune liver tissue damage. Methods： The syngeneic liver antigen emulsified in complete Freund＇s adjuvant was injected intraperitoneally into adult female C57BL/6 mice twice. Liver injury was assessed by serum ALT, AST and liver histopathological changes. The autoantibody was detected by immunofluorescence. Using real-time quantita- tive-polymerase chain reaction （Realtime-PCR） and immunohistochemisty to detect gene transcription and protein expression of Fas and FasL in the mouse liver, and cell apoptosis was detected in liver tissue by TUNEL. Results： In liver tissue of model mice after 28-days modeled, Fas and FasL mRNA and protein levels which mainly expressed in liver ceils and infiltrating lympho- cytes were significantly enhanced （ P 〈 O. 05 ） . No abnormal changes in normal mouse liver tissue while model mice showed scattered or focal infiltration of lymphocytes by light microscopy. The TUNEL-positive hepatocytes and lymphocytes can be seen in the inflammatory regional centers and around, and apoptosis index （AI） in the liver tissue of the model group were significantly elevated. Conclusion： Inflammatory infiltration and apoptosis coexist in liver tissue of experimental autoimmune hepatitis. Cell apoptosis mediated by the Fas / FasL path is associated with the development of autoimmune hepatitis.

关 键 词：自身免疫性肝炎 FAS FASL 凋亡 小鼠 

分 类 号：R575.1[医药卫生—消化系统]