'益肝康'大鼠药物血清对HSCs c-fos、c-jun基因表达的影响

摘要

目的 观察"益肝康"药物血清对HSCs c-fos、c-jun基因表达的影响,试图从基因转录水平探讨"益肝康"的抗肝纤维化作用机制.方法 制备肝纤维化模型鼠,造模成功后,药物组以每天20 ml/kg剂量分2次灌服"益肝康";对照组灌以等量0.9%氯化钠溶液.连续给药6 d后经下腔静脉取血并分离血清.采用盲法,用上述10%药物血清培养HSCs 24 h后,采用RT-PCR技术检测c-fos、c-jun mRNA的表达情况.结果 ①电泳条带的光密度扫描显示,B、D组c-fos mRNA的表达与C组比较有差异有统计学意义(P＜0.05);B、D组与A组比较,c-fos mRNA的表达亦明显下降(P＜0.05);② 光密度扫描显示,B、D组c-jun mRNA的表达同C组比较,差异有统计学意义(P＜0.05);B、D组与A组比较,c-jun mRNA的表达亦明显受到抑制(P＜0.05).结论 活化的HSCs表达较高水平的c-fos与c-jun mRNA,表明c-fos与c-jun立早基因可能参与了其增殖过程;"益肝康"药物血清均能显著下调c-fos与c-jun mRNA的表达,在转录水平对c-fos与c-jun的调节可能是两者发挥其抗肝纤维化作用靶点之一.%Objective To investigate the effect of medicated serum with Yigankang decoction on the expression of c-fos and c-jun in HSCs in order to explore the action mechanism of anti-hepatic fibrosis of Yigankang at genetic transcription level. Methods After the rats were modeled into hepatic fibrosis, Yigankang was given by gavage for the rats in every group except for control group in which the same volume 0.9％ NS was given twice a day for 6 days. Then the pharmacological serum was extracted from the inferior vena cava of the rats. Mter 24h incubation with related serum that was added blindly to HSCs,the expressions of c-fos mRNA and c-jun mRNA in HSCs were detected by reverse transcription polymerase chain reaction (RT-PCR). Results ① Yigankang decoction medicated serum extracted from normal rats and CC14-induccd liver fibrosis rats could inhibit c-fos mRNA expression in HSCs. As compared with HSCs treated by serum from CC14-induced liver fibrosis rats, the expression of c-fos mRNA in HSCs treated by medicated serums with Yigankang decoction was significantly decreased ( P ＜ 0.05 ). As compared with HSCs treated by serum from normal rats, the expression of c-fos mRNA in HSCs treated by medicated serums with Yigankang decoction was significantly decreased ( P ＜ 0.05 ). ②Yigankang decoction medicated serum extracted from normal rats and CCl4-induced liver fibrosis rats could inhibitn c-jun mRNA expression in HSCs. As compared with HSCs treated by serum from CC14-induced liver fibrosis rats,the expression of c-jun mRNA in HSCs treated by medicated serums was significantly decreased( P ＜ 0.05 ). As compared with HSCs treated by serum from normal rats, the expression of c-jun mRNA in HSCs treated by medicated serums was significantly decreased ( P ＜ 0.05 ). Conclusion Yigankang decoction medicated serum extracted from normal rats and CCl4-induced liver fibrosis rats can inhibit the expression of c-fos and c-jun mRNA during liver fibrosis pathogenesis.