大黄素对肝星状细胞增殖及细胞外信号调节激酶的影响

目的:观察大黄素对HSC-T6细胞增殖及细胞外信号激酶(ERK1)表达的影响。方法:将培养细胞分成正常组和大黄素不同浓度干预组(大黄素终浓度分别为20μmol/L、40μmol/L、60μmol/L),采用MTT法观察大黄素对HSC-T6增殖的影响;采用流式细胞仪观察各组细胞周期的变化;采用反转录聚合酶链式反应(RT-PCR)半定量法观察ERK1mRNA的表达;采用免疫细胞化学SABC法,观察HSC-T6 ERK1表达的变化。结果:不同浓度大黄素对HSC-T6的增殖均有抑制作用(与正常组比较P0.05,P0.01),且呈一定的量效关系。大黄素作用HSC-T6细胞后,G0/G1期增加,S期减少;当浓度为40～60μmol/L时,G2/M期减少(与正常组比较均P0.05)。正常组HSC-T6有ERK1mRNA表达,而大黄素使ERK1mRNA的表达显著降低,(与正常组比较P0.05,P0.01)。正常组HSC-T6 ERK1阳性均强表达,大黄素使ERK1阳性表达细胞数量减少,阳性减弱。结论:大黄素对HSC-T6细胞增殖有抑制作用,其作用与阻滞细胞于G0/G1期、抑制其ERK1表达有关。

Effect of Emodin on the Cell Proliferation and Extracellular Signal-regulated Kinase in Hepatic Stellate Cell-T6

Objectives: To explore the effect of emodin on the cell proliferation and extracellular signal-regulate kinase in hepatic stellate cell-T6.Methods:The immortalized rat hepatic stellate cell line HSC-T6 was commonly cultured after revival,and was allocated into normal control group and treated with different concentrations emodin.The cell proliferation was assessed via MTT assay,and cell cycle was analyzed by flow cytometry,and the expression of ERK1 mRNA was measured by semi-quantitative RT-PCR,and the expression of ERK1 by immunohistochemistry.Results: After intervention of different concentration emodin(the extreme concentrations are 20μmol/mL,40μmol/mL,60μmol/mL respectively),the inhibition rates of proliferation increased significantly(compared with the normal group,P<0.05,P<0.01) and had dose-dependent manner.In cell cycle progression,emodin can inhibit cell cycle and the cell at G0/G1 phase increase obviously but descend at S phase,and G2/M phase decrease when emodin concentration is at 40~60μmol/mL(compared with the normal group,P<0.05).After exposure to emodin,the expressions of ERK1 mRNA are obviously downregulated,and there is statistical significance compared with the normal group(P<0.05,P<0.01).Immunohistiochemical results showed that the position cells ERK1 positive expression reduced obviously and the strength decrease.Conclusion: Emodin can inhibit the proliferation of HSC-T6,depress cell cycle at G0/G1 phase and restrain the expressions of ERK1 and ERK1 mRNA in HSC-T6.