【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。更多还原

【Abstract】 Objective: To establish experimental autoimmune hepatitis(EAH) mouse model and observe Fas /FasL expression 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 quantitative-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 cells and infiltrating lymphocytes were significantly enhanced(P < 0.05).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.更多还原