槲皮素对过氧化氢诱导的人脐静脉内皮细胞AKT/mTOR信号通路及自噬的影响

目的：分析槲皮素对人脐静脉内皮细胞自噬和增殖的影响，探讨其与AKT/mTOR信号通路调控的关系及机制。方法：培养人脐静脉内皮细胞，MTT法检测细胞活力，MDC染色法检测自噬泡水平，Western印迹检测自噬相关蛋白水平，运用3-MA研究槲皮素对自噬和细胞增殖的影响。结果：槲皮素能增加人脐静脉内皮细胞活力(P<0.01),上调细胞中MDC染色标记的荧光颗粒，并伴随浓度的上调明显增加(P<0.01),能促进脐静脉内皮细胞中自噬蛋白LC3Ⅱ/LC3Ⅰ水平表达(P<0.01),降低p-AKT/AKT、p-mTOR/mTOR水平(P<0.01)。与槲皮素组比较，槲皮素+3-MA组细胞活力和LC3Ⅱ/LC3Ⅰ均明显降低(P<0.05),p-mTOR/mTOR明显增加(P<0.05)。结论：槲皮素可通过抑制AKT/mTOR信号通路诱导细胞自噬来调控脐静脉内皮细胞增殖。

Effect of quercetin on hydrogen peroxide-induced AKT/mTOR signaling pathway and autophagy in human umbilical vein endothelial cells

ObjectiveTo analyze the effect of quercetin on autophagy and proliferation of human umbilical vein endothelial cells and its relationship with AKT/mTOR signaling pathway.MethodsMTT method was used to detect cell viability.MDC staining was used to detect the level of autophagy vesicles.Western blotting was used to detect the levels of autophagy-related proteins, and 3-MA was used to study the effect of quercetin on autophagy and cell proliferation.ResultsQuercetin increased the viability of human umbilical vein endothelial cells and up-regulated the fluorescent particles labeled by MDC staining in the cells followed with the increase in concentration, which promoted the expressions of autophagy protein LC3 Ⅱ/LC3 Ⅰ in umbilical vein endothelial cells and reduce the levels of p-AKT/AKT and p-mTOR/mTOR(P < 0.01).Compared with the quercetin group, the cell viability and LC3 Ⅱ/LC3 Ⅰ of the quercetin+3-MA group were significantly reduced, and p-mTOR/mTOR was significantly increased(P < 0.05).ConclusionsQuercetin can regulate the proliferation of umbilical vein endothelial cells by inhibiting the AKT/mTOR signaling pathway and inducing autophagy.