淫羊霍苷对HCY诱导的血管内皮细胞内质网应激的影响

目的:研究淫羊藿苷(icariin,ICA)对同型半胱氨酸(homocysteinemia,HCY)诱导的血管内皮细胞内质网应激的影响,探讨ICA抗动脉粥样硬化的机制。方法:建立HCY诱导的兔血管内皮细胞损伤模型,与不同浓度的ICA共同培养48h后,TUNEL法检测细胞凋亡率;RT-PCR检测葡萄糖调节蛋白78(glucose regulated protein78,GRP78)基因表达;目的基因克隆、鉴定并测序。结果:荧光显微镜观察显示HCY模型组可见多数凋亡细胞,ICA组细胞未见明显凋亡形态学改变;ICA处理组细胞GRP78mRNA表达明显减少,与HCY组比较差异显著(P0.01);测序结果表明,来源于血管内皮细胞的全长648bp的基因片断与兔GRP78基因高度同源。结论:ICA拮抗HCY诱导的血管内皮细胞凋亡,减缓血管内皮细胞内质网应激可能是其作用机制之一。

Effects ofICA on HCY-induced Vascular Endothelial Cells Endoplasmic Reticulum Stress

Objective: To study the effects of ICA on HCY-induced vascular endothelial cells endoplasmic reticulum stress, and to explore ICA anti-atherosclerotic mechanisms. Method: HCY-induced rabbit endothelial cell injury models were established, and co-cultured with different concentrations of ICA for 48 hours, then apoptosis rate was assayed by TUNEL ; Glucose-regulated protein 78 (GRP78) gene expression was done by RT-PCR detection; The wanted gene was cloned, identified and sequenced. Result: The fluorescence microscope observation showed that there were majority of apoptotic cells in HCY model group, and in ICA group there was no significant morphological changes of apoptosis. Compared with HCY group, cell GRP78 mRNA expression in ICA treatment group significantly reduced (P<0.01). Sequencing results showed that full-length 648bp gene fragment derived from vascular endothelial cells was highly homologous with rabbit GRP78 gene. Conclusion: ICA can resist HCY-induced endothelial cell apoptosis and slow down vascular endothelial cells endoplasmic reticulum stress, which may be one of the mechanisms of its role.