罗格列酮对肝星状细胞的生物学特性及PDGF表达的影响

Effect of rosiglitazone on biological characteristics of hepatic stellate cells and their expression of PDGF

目的观察罗格列酮对体外培养的肝星状细胞(hepatic stellate cells,HSC)增殖、凋亡、细胞周期、超微结构及表达α-SMA、PDGF-B的影响。方法 MTT法检测罗格列酮对HSC增殖的影响;流式细胞仪(FCM)检测罗格列酮对HSC细胞周期和凋亡率的影响;免疫组化SABC法检测罗格列酮对HSC表达α-SMA、PDGF-B的影响;电镜观察罗格列酮对HSC超微结构的影响。结果 MTT法检测显示罗格列酮对HSC的增殖具有抑制作用,抑制率随着罗格列酮浓度的增加而增加;FCM检测:罗格列酮作用后,HSC的凋亡率、G0/G1期细胞比例均随着浓度的增加而增加,各浓度组凋亡率的增加均具有显著性意义(P<0.01);免疫组化染色显示HSC中α-SMA、PDGF-B的表达水平随着罗格列酮浓度的增加而减少,且两者的表达具有紧密相关性(r=0.971);电镜观察显示HSC的生长状态受到抑制,部分细胞呈现凋亡相关的超微结构改变。结论罗格列酮能抑制HSC的增殖、活化,其作用机制可能与阻滞HSC于G0/G1期、诱导HSC凋亡及降低HSC的PDGF表达有关。

Purpose To observe the influence of rosiglitazone on ultrastructure,proliferation,apoptosis and cell circle of hepatic stellate cells（HSC） in vitro,and to study the effect of rosiglitazone on the expression of α-SMA and PDGF-B in HSC.Methods MTT assay was used to detect the influence of rosiglitazone on the proliferation of HSC,flow cytometry（FCM） was employed to investigate the effect of rosiglitazone on the ratio of apoptosis and the distribution of cell cycle,and the expression levels of α-SMA and PDGF-B in HSC treated by rosiglitazone were semi-quantitatively analyzed by IHC.Transmission electron microscope（TEM） was applied to observe the ultrastructural change of HSC after treatment by rosiglitazone.Results MTT results showed that rosiglitazone could inhibit the proliferation of HSC.Inhibitory rate increased with increasing rosiglitazone concentration in a dose-effect manner,FCM results showed that both apoptosis rate and the percentage of G0/G1 phase increased with increasing rosiglitazone concentration.The increase of apoptosis rate in each group was very significant（P0.01）.IHC results showed that： expression of α-SMA and PDGF-B in HSC was decreased with the increase of rosiglitazone concentration,and the expression of both antigens was closely related（r=0.971）.TEM results showed that after rosiglitazone treatment（15 μmol/L）,growth state HSC was inhibited,and a part of HSC presented the ultrastructural changes of apoptosis.Conclusion Rosiglitazone could inhibit the proliferation and activation of HSC.The mechanism could be related with arresting HSC in the G0/G1 phase,inducing the apoptosis of HSC and decreasing the expression of PDGF in HSC.