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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P<0.05,P<0.01),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P<0.05)。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P<0.05,P<0.01)。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。
Objective: To observe the effects of emodin on the proliferation and the expression of extracellular signal kinase (ERK1) in HSC-T6 cells. Methods: The cultured cells were divided into normal group and different concentrations of emodin intervention group (final concentrations of emodin were 20μmol / L, 40μmol / L, 60μmol / L). The effects of emodin on the proliferation of HSC-T6 were observed by MTT method. The changes of cell cycle in each group were observed by flow cytometry. The expression of ERK1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) semi-quantitative method. The expression of ERK1 was observed by immunocytochemical SABC method. Results: Different concentrations of emodin inhibited the proliferation of HSC-T6 (P <0.05, P <0.01 compared with the normal group), and showed a certain dose-effect relationship. Emodin increased the number of G0 / G1 phase and decreased the number of S phase in HSC-T6 cells. When the concentration of 40-60 μmol / L, G2 / M phase decreased (all P <0.05 compared with the normal group). The normal group HSC-T6 ERK1mRNA expression, and emodin ERK1mRNA expression was significantly reduced (compared with the normal group P <0.05, P <0.01). In normal group, the expression of ERK1 in HSC-T6 was strongly positive. Emodin reduced the number of ERK1-positive cells and decreased the expression of ERK1. CONCLUSION: Emodin can inhibit the proliferation of HSC-T6 cells and its effect is related to arresting cells in G0 / G1 phase and inhibiting the expression of ERK1.