平阳霉素对人脐静脉内皮细胞株EA.hy926的作用及其机制

目的:观察平阳霉素(PYM)对人脐静脉内皮细胞株EA.hy926的作用及可能机制。方法:采用甲基三氯硅烷(MTS)比色法检测PYM对EA.hy926细胞体外生长的作用;流式细胞术测定PYM作用细胞24h后细胞凋亡、半胱氨酸天冬氨酸蛋白酶(Caspase)-3表达、线粒体膜电位的变化;免疫印迹(Western-blot)法检测PYM对EA.hy926细胞中Caspase-8、Caspase-9蛋白表达水平的影响。结果:PYM可明显抑制EA.hy926细胞的增殖,且生长抑制率具有浓度时间依赖性;流式细胞术检测结果显示PYM各浓度作用24h后,细胞凋亡百分率随药物浓度增加而增高,活化Caspase-3表达在PYM各浓度组均增强,线粒体膜电位在各浓度组均下降;Western-blot法检测结果显示PYM作用EA.hy926细胞后,Caspase-8、Caspase-9蛋白均可出现明显的剪切带。结论:PYM能抑制EA.hy926细胞的增殖,这种抑制作用是可通过诱导细胞凋亡实现的,Caspase-3、Caspase-8、Caspase-9参与了PYM诱导EA.hy926细胞凋亡的过程。

The effect of Pingyangmycin on cultured human umbilical vein endothelial cells line FA.hy926 and its mechaism

Objective: To observe the effect and mechanism of Pingyangmycin(PYM)on cultured human umbilical vein endothelial cells line EA.hy926.Methods: The inhibitory rates of EA.hy926 cells,which were treated with PYM for 24 h and 48 h,were compared respectively by MTS assay.The changes in the apoptosis,Caspase-3 protein expression and mitochondrial transmembrane potential of the cells managed with PYM for 24 h were examined with flow cytometry(FCM).Results: The results of MTS colourimetry showed that the proliferation of EA.hy926 cells dealed with PYM for 24 h and 48 h could markedly be inhibited,and the growing inhibition ratio was concentration-time-dependent.FCM results showed that the apoptosis rate and the expression of active caspase-3 protein were also increased in every group of different concentration,and mitochondrial transmembrane potential was declined with the increasing of concentration of drug.Western-bolt results showed that the shearing band were showed up in caspase-8,caspase-9 protein.Conclusion: PYM can possibly inhibit the growth of EA.hy926 cells,and the inhibition may include apoptosis induction.Caspase-3,caspase-8,caspase-9 may play an important role in the induced apoptosis.