淫羊藿苷对缺氧诱导血管内皮细胞损伤的保护作用

Effect of Icariin on Hypoxia Induced Vascular Endothelial Cells Injury

目的研究淫羊藿苷(icariin,ICA)对缺氧所致血管内皮细胞(vascularendothelialcells,VECs)损伤的影响。方法建立体外细胞缺氧模型,用MTT法观察ICA对缺氧损伤的血管内皮细胞活性的影响,测定细胞中丙二醛(MDA)含量、超氧化物歧化酶(SOD)及乳酸脱氢酶(LDH)活力,并对细胞进行Hoechst33342荧光染色、电镜观察细胞超微结构,运用流式细胞仪及DNA琼脂糖凝胶电泳分析DNA断裂情况,以观察ICA对缺氧诱导的内皮细胞凋亡的影响。结果ICA能抑制缺氧引起的血管内皮细胞的减少,降低LDH活力,并抑制缺氧条件下MDA生成,提高SOD活力;缺氧能诱导血管内皮细胞凋亡,表现为细胞核浓缩,沿核膜排列成块状,DNA琼脂糖凝胶电泳显示典型“梯形条带”,流式细胞仪分析呈现典型凋亡亚二倍体峰。ICA能显著抑制缺氧诱导的内皮细胞凋亡。结论ICA具有保护缺氧诱导的血管内皮细胞损伤的作用,其作用机制与抗脂质过氧化物产生、提高SOD活力以及抗细胞凋亡有关。

ObjectiveTo study the effect of icariin on vascular endothelial cells (VECs) injury induced by hypoxia. MethodsThe hypoxia-ischemia model was established. The effect of icariin on injury of VECs activity induced by hypoxia was determined by MTT assay. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and lactate dehydrogenase (LDH) activity in cell homogenate were measured with corresponding kit. Effect of icariin on cells apoptosis induced by hypoxia was determined by Hoechst 33342 fluorescent staining, cell ultrastructure observation under transmission electron microscopy and analysis on gene fragmentation by flow cytometry and DNA gel electrophoresis. ResultsICA could inhibit the hypoxia induced VECs reduction, suppress LDH activity, reduce the MDA production, and enhance SOD activity under hypoxia.Hypoxia could induce VECs apoptosis, revealed chromation condensed in nuclei with the fragments arranged along the nuclear membrane. DNA gel electrophoresis showed typical ladder strands of DNA. Cells displayed a typical sub-diploid peak in flow cytometry. ICA could significantly inhibit the hypoxia induced apoptosis of VECs. ConclusionICA has the protective effect on hypoxia injured VECs, which may be related to its effect of anti-apoptosis, anti-lipid peroxidation and SOD activity enhancing.