消退素E1对氧化型低密度脂蛋白诱导的血管内皮细胞损伤的保护作用

Resolvin E1 protects against ox-LDL-induced injury on vascular endothelial cells

目的研究消退素E1是否对氧化型低密度脂蛋白诱导的人脐静脉内皮细胞损伤有保护作用，并探讨其分子机制。方法将人脐静脉内皮细胞（HUVEC）随机分成6组，分别用生理盐水、消退素E1、P13K抑制剂wortmanin、氧化型低密度脂蛋白、氧化型低密度脂蛋白＋消退素E1、氧化型低密度脂蛋白＋消退素E1＋PBK抑制剂wortmanin处理人脐静脉内皮细胞48h。随后用噻唑蓝法分析内皮细胞存活率、流式细胞仪检测细胞凋亡率、酶联免疫法分析细胞上清中肿瘤坏死因子-α（TNF-α）含量，酶标仪测量细胞内半胱天冬蛋白酶（caspase）3、9的活性，免疫印迹法检测激活的AKT和血凝素样氧化低密度脂蛋白受体-1（LOX-1）的表达。结果氧化型低密度脂蛋白组细胞活力显著低于生理盐水组（P〈0．01），细胞凋亡率、TNF-α含量、caspase3和9活性，LOX-1蛋白的表达均显著高于生理盐水组（P均〈0．01）。氧化型低密度脂蛋白＋消退素E1组细胞凋亡率、TNF．d含量、caspase3和9活性，LOX．1蛋白的表达均显著低于氧化型低密度脂蛋白组（P〈0．01），细胞活力和激活的AKT均显著高于氧化型低密度脂蛋白组（p均〈0．01）。氧化型低密度脂蛋白＋消退素E1＋P13K抑制剂wortmanin组的细胞活力和细胞凋亡率、TNF-α含量、caspase3和9活性，LOX-1蛋白的表达均显著高于氧化型低密度脂蛋白＋消退素E1组（P〈0．05）。结论消退素E1能有效地抑制氧化型低密度脂蛋白诱导的内皮细胞凋亡，此作用可能通过P13K／Akt信号通路介导。

Objective To investigate whether Resolvin E1 （ RvE1 ） could protect against ox-LDL- induced injury on human vein vascular endothelial cells and reveal related molecular mechanisms. Methods Human vein vascular endothelial ceils were randomly assigned to six groups, which were treated with saline, RYE1, wortmanin, ox-LDL, ox-LDL and RvE1, ox-LDL and RvE1 and wortmanin, respectively. After 48 h, survival rates were determined by MTI＇, apoptosis rate of cells were determined by flow cytometry, TNF-ct contents were assayed by ELISA, caspase 3 and 9 activities were measured by microplate reader, and the expression of p-AKT and LOX-1 were determined by Western blot. Result Compared with normal saline group, survival rate was markedly decreased and apoptosis rate, TNF-α content, caspase 3 and 9 activities, and the expression of LOX-1 were significantly increased in ox-LDL group （ P 〈 0. 01 ）. Survival rate was significantly increased and apoptosis rate, TNF-α content, caspase 3 and 9 activities, and the expression of LOX-1 were significantly decreased in ox-LDL ＋ RvE1 group compared to ox-LDL group （ P 〈 0.01 ）, these beneficial effects of RyE1 could be blocked by PI3K inhibitor wortmanin （ P 〈 0. 05 ）. Conclusion The present data showed that RvE1 could effectively protect against ox-LDL-induced endothelial cell injury, which might be mediated by PI3 K-AKT signaling pathway.