【摘要】 目的:探讨丹参单体IH764-3对H2O2刺激的肝星状细胞(HSCs)增殖、凋亡等细胞行为的影响及细胞外信号调节激酶1(ERK1)在其中的调节作用。方法:应用体外细胞培养技术,采用直接细胞计数法、3H-胸腺嘧啶核苷(3H-TdR)掺入法测定HSCs增殖;透射电镜、膜联蛋白(Annexin-V)/碘化丙啶(PI)双标记流式细胞术测定HSCs凋亡;分别应用Western blot和逆转录-聚合酶链反应(RT-PCR)技术测定ERK1蛋白及其mRNA的表达。结果:①H2O2具有刺激HSCs增殖的作用;②丹参单体IH764-3剂量依赖性抑制H2O2刺激的HSCs增殖;③Annexin-V/PI检测显示,10 mg/L,20 mg/L,30 mg/L及40 mg/L IH764-3干预48 h后各组凋亡率分别为6.35%、9.28%、15.10%、19.69%,而H2O2组为2.30%;30 mg/L IH764-3干预HSCs不同时间(12 h、24 h、48 h)的凋亡率分别是6.73%、10.34%、15.10%,呈时间依赖性;④丹参单体IH764-3干预组,HSCs的ERK1蛋白及其mRNA表达下调。结论:丹参单体IH764-3可以抑制HSCs增殖并诱导其凋亡;这种作用与其抑制ERK1蛋白和ERK1mRNA表达有关。更多还原

【Abstract】 Objective: To explore the effect of Salvia miltiorrhiza monomer IH764-3 on apoptosis in hydrogen peroxide(H2O2)-stimulated hepatic stellate cells(HSCs).Methods: HSCs were cultured in medium with different IH764-3 doses(10 mg/L,20 mg/L,30 mg/L,40 mg/L) and without IH764-3.Direct cell count,3H-thymidine incorporation,Annexin-V/Propidium Iodide double-labeled flow cytometry, TUNEL and transmission electron microscopy were employed to estimate the influence of IH764-3 on proliferation and apoptosis of HSCs.The expression of extracellular signal-regulated kinase 1(ERK1) mRNA and protein in HSCs were detected using RT-PCR and Western blot respectively. Results: It was showed that H2O2 could promote HSC proliferation.In contrast,IH764-3 at concentrations of 10 mg/L,20 mg/L,30 mg/L and 40 mg/L inhibited its proliferation.The inhibition rates were 7.13%,28.36%,53.80% and 73.10%(P<0.01).And the inhibition rates of IH764-3 at concentrations of 30 mg/L at 12 h,24 h and 48 h were 22.24%,40.51% and 61.65%.Furthermore,IH764-3 could also induce the HSC apoptosis in dose-dependent and time-dependent manners(P<0.01).In addition,after exposed of HSCs to IH764-3 for 24 h,ERK production decreased and ERK1 mRNA was down-regulated earlier about 2 h after exposure to IH764-3.Conclusion: IH764-3 may inhibit the proliferation and induce apoptosis of HSCs in both dose-dependent and time-dependent manners,which may be related to down-regulation of ERK expression.更多还原