转化生长因子β_1在白细胞介素10干预肝纤维化大鼠肝星形细胞的表达  

Expression of Transforming Growth Factor-β_1 of Hepatic Stellate Cells of Hepatic Fibrosis Rat after Interleukin-10 Intervention

作　　者：史美娜[1] 郑伟达[1] 张丽娟[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化科,福州350001

出　　处：《福建医科大学学报》2004年第4期377-380,共4页Journal of Fujian Medical University

基　　金：福建省科技开发计划项目资助课题 ( 2 0 0 3D0 5 )

摘　　要：目的　探讨转化生长因子 β1(TGF β1)在白细胞介素 10 (IL 10 )干预肝纤维化大鼠星形细胞中的表达 ,阐明外源性IL 10的抗纤维化作用及可能机制。　方法　 6 0只雄性SD大鼠随机分为正常对照组 (N组 ,8只 )、肝纤维化模型组 (C组 ,2 8只 )和IL 10干预组 (I组 ,2 4只 ) ,分别于CCl4造模第 7周及 11周采用链霉蛋白酶E、Ⅳ型胶原酶经门静脉灌注分离肝星形细胞。半定量RT PCR法检测各组新分离肝星形细胞中TGF β1mRNA的表达水平 ;SP免疫细胞化学法检测各组原代培养 72h后肝星形细胞TGF β1蛋白的表达水平。并于上述两个时间段分别将各组肝组织行苏木精 伊红染色判断炎症和肝纤维化程度。　结果　成功建立大鼠肝纤维化模型及IL 10干预模型 ,并对大鼠肝星形细胞进行分离及原代培养。与正常组相比 ,纤维化模型组TGF β1的mRNA水平显著升高(P <0 0 1) ,经IL 10干预后则明显降低 (P <0 0 1)。纤维化模型组星形细胞TGF β1mRNA水平在第 11周时低于第 7周 (P <0 0 5 )。SP免疫细胞化学法检测各组TGF β1的蛋白表达情况与mRNA结果相平行。　结论　IL 10可抑制肝纤维化大鼠星形细胞中TGF β1表达 ,具有抗纤维化作用。Objective To investigate the expression of transforming growth factor-β-1(TGF-β-1) of hepatic stellate cells(HSC) of hepatic fibrosis rat after treatment with interleukin-(IL-10) and the antifibrogenic role of IL-10. Methods Hepatic fibrosis was induced by CCl-4. Sixty male Spraque-Dawly(SD) rats were randomly divided into three groups: normal control group(group N, 8 rats), hepatic fibrosis model group(group C, 28 rats) and IL-10 treated group(group I, 24 rats). At the beginning of the 7th and 11th week, rats in each group were routinely perfused with pronase E and type Ⅳ collagnase through portal vein catheter and the suspension obtained from the digested liver was centrifuged with 11% nycodenz density gradient to isolate HSC. RT-PCR was used to analyze TGF-β-1 mRNA. Immunocytochemistry was performed to detect protein expression in cultured HSC. Results Hepatic fibrosis was developed with the increase of the injection frequency of CCl-4 and HSC were successfully isolated. At the 7th week, TGF-β-1mRNA increased in group C compared to group N(P<0.01), and significantly decreased after IL-10 intervention(P<0.01). At the 11th week, TGF-β-1mRNA in intervention group was still lower than CCl-4group(P<0.01). The hepatic fibrosis immunochemistry were paralleled with TGF-β-1 expression. Conclusion The expression of TGF-β-1 increased in the early stage of hepatic fibrosis but decreased by IL-10 intervention. IL-10 play an antifibrogenic role in suppressing TGF-β-1 expression.

关 键 词：肝 星形细胞 肝硬化 白细胞介素10 转化生长因子Β1 大鼠 

分 类 号：R575.2[医药卫生—消化系统]