肝纤维化大鼠星状细胞Smad3、Smad7基因表达差异的研究

Study of Different Expression of Smad3 and Smad7 Genes in the Hepatic Stellate Cells of Rat with Liver Fibrosis

通过检测肝纤维化大鼠原代肝星状细胞 (HSC)的 Sm ad3和 Smad7编码基因的表达 ,探讨 HSC活化及转化机制。皮下注射四氯化碳 (CCl4)制备大鼠肝纤维化模型 ,于注射 CCl4后 1、 4、 8周分批分离 HSC并计算 HSC平均得率 ;RT- PCR检测其 Sm ad3、Smad7m RNA的表达变化。结果显示 :1CCl4注射后第 1、 4、 8周大鼠 HSC平均得率分别为 :(3.34± 0 .36 )× 10 7/只、 (4 .2 1± 0 .6 4)× 10 7/只、 (5 .88± 0 .79)× 10 7/只 ,均高于正常大鼠 HSC平均得率(2 .2 3± 0 .19)× 10 7/只 ,组间比较有显著性差异 ,P<0 .0 5。 2与正常对照组相比 ,注射 CCl4后 1、 4、 8周大鼠 HSC中 Sm ad3m RNA表达增加 ,P<0 .0 5。注射 CCl4后 1周 Sm ad7m RNA较正常明显升高 ,P<0 .0 5 ,到第 4周和第 8周表达水平则持续下降 ,P<0 .0 1。提示 Sm ad3和 Smad7在肝纤维化发生、发展中对 HSC的活化及转化起不同的介导作用。

To evaluate the mechanism of activation of hepatic stellate cell (HSC) and its phenotype trans-differentiation, the expression of Smad3 and Smad7 gene in primary HSC of rat with liver fibrosis was detected. The liver fibrosis model of rat was made by subcutaneously injected with CCl 4 . HSCs were isolated at 1st , 4th and 8th week after injection of CCl 4 . The average yield of HSC was calculated. PT-PCR analysis was carried to detect the changes of Smad3 and Smad7 mRNA expression in HSCs. The results showed that the yield of HSC was (3.34±0.36)×10 7, (4.21±0.64)×10 7 and (5.88±0.79)×10 7 in rats respectively at 1st, 4th and 8th week after injection of CCl 4, all significantly higher than in the normal rats [(2.23±0.19)×10 7, P<0.05]. Compared to normal control group, the level of Smad3 mRNA expression in HSCs of fibrosis rats was increased significantly. However, Smad7 mRNA changed in the other way. Higher level was observed one week after injection of CCl 4 , but lower level seen from 4 weeks after injection of CCl 4 to the end (P<0.01). It indicated that there were different roles between Smad3 and Smad7 in activation of HSC during liver fibrogenesis.