Objective To observe two main subtypes of MAPK family extracellular signal-regulated kinase(ERK) and C-Jun-N-terminal kinase (JNK) activation in HSCs, and to investigate the effects of medicated serum with Yigankang on the two proteins activation in HSCs. Methods After the rats were prepared into hepatic fibrosis models,the rats were randomly divided into 4 groups, 8 rats in each group, group B and group D were administration groups, and group A and group C were control group. Yigankang was given by gavage to administration groups,however,the same volume 0.9％ NS was given to control groups twice a day for 6 days,then the serum with drugs was extracted from the inferior vena cava of rats. The HSCs were cultured with 10％medicated serum medium for 24h blindly, then the expressions of P-ERK and P-JNK were detected by Western Blot respectively. Results Western Blot analysis showed that there were zones of hybridization in the position of 42,44 KD in every group,which indicated that activated HSC s expressed P-ERK1 and P-ERK2 simultaneously ;there were zones of hybridization in the position of 46,54 KD in every group,which indicated that activated HSCs expressed P-JNKland P-JNK2 simultaneously. After the intervention of medicated serum with Yigankang,the expressions of P-ERK and P-JNK in HSCs were decreased significantly,as compared with those in control groups ( P ＜0.05).Conclusion ERK and JNK keep higher phosphorylation levels in activated HSCs, which suggests the signal transduction way mediated by the two enzymes is one of the important ways that activate and proliferate HSCs, while the block of MAPK signal way may be one of important ways that Yigankang treat liver fibrosis.%目的 探讨肝星状细胞丝裂原活化蛋白激酶(HSCs MAPK)通路中信号调节蛋白激酶(ERK)、C-Jun氨基末端蛋白激酶(JNK)的活化情况,并进一步研究益肝康药物血清对P-ERK、P-JNK蛋白表达的影响.方法制备肝纤维化模型鼠.分为正常大鼠血清干预组(A组);正常大鼠益肝康药物血清干预组(B组);肝纤维化模型大鼠血清干预组(C组);肝纤维化大鼠益肝康药物血清干预组(D组).每组8组,B、D组为药物组,A、C组为对照组.造模毕,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Western blot技术检测各组HSCs P-ERK、P-JNK蛋白表达. 结果 Western blot分析结果显示,各组在大概42、44 KD的位置上均出现杂交带,表明活化的HSCs同时表达P-ERK1和P-ERK2;各组在大概46、54 KD的位置上均出现杂交带,表明HSCs同时表达P-JNK1和P-JNK2.经肝纤维化大鼠益肝康药物血清干预后,P-ERK、P-JNK蛋白表达水平较病理对照组显著降低(P<0.01);正常大鼠益肝康药物血清干预后,P-ERK、P-JNK蛋白表达水平与正常大鼠血清干预组比较,也均显著减少(P<0.05).结论 活化的HSCs中ERK、JNK保持着较高的磷酸化水平.提示两者介导的信号转导通路是HSCs活化和增殖的重要途径之一;阻断MAPK信号通路可能是益肝康治疗肝纤维化的重要作用途径之一.
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