银杏内酯B抑制高糖诱导内皮细胞凋亡及机制研究

摘要

目的 探讨银杏内酯B对高糖诱导的内皮细胞凋亡的影响及相关的分子机制.方法 使用脐静脉血管内皮细胞作为细胞模型,分为对照组、高糖组、银杏内酯B处理组.用Transwell实验检测内皮细胞迁移,AnnexinⅤ试剂盒检测细胞凋亡,荧光试剂盒检测活性氧(reactive oxygen species,ROS)水平,Western blot检测Bax,Bcl-2,caspase-3的表达以及p53的磷酸化.结果 高糖(30 mmol·L-1)处理组内皮细胞迁移数量明显降低,迁移率为(66.6±15.6)%,而银杏内酯B(0.6 g·L-1)处理增加了内皮细胞的迁移数量,迁移率为(94.8±11.4)%.高糖处理导致内皮细胞ROS水平增高2.75倍,与高糖处理组比较,0.6 g·L-1银杏内酯B完全抑制了高糖诱导的ROS产生.流式细胞仪分析表明,高糖刺激内皮细胞凋亡率为(53.8±2.6)%,银杏内酯B处理组内皮细胞凋亡率为(44.0±3.1)%.对凋亡相关蛋白的分析表明,高糖组Bax、caspase-3表达增加,Bcl-2表达降低,银杏内酯B处理抑制了Bax、caspase-3表达,并增加了Bcl-2表达.此外,高糖处理增加了内皮细胞p53表达及磷酸化,而银杏内酯B抑制高糖刺激的p53活化.结论 银杏内酯B能抑制高糖诱导的内皮细胞凋亡,改善内皮细胞迁移功能,对高糖引起的内皮细胞损伤具有保护作用.%Aim Toinvestigatetheeffectofginkgolide B on apoptosis in high glucose-treated endothelial cells.Methods Humanumbilicalveinendothelial cells(HUVECs)were used in the present study.The level of transmigration of HUVECs was analyzed by Tr-answell experiment.Apoptosis was detected by flow cy-tometry.Reactive Oxygen Species (ROS ) was meas-ured by immunofluorescence kit.The protein expres-sionwasanalyzedbyWesternblot.Result Highglu-cose treatment resulted in a reduction in transmigration of HUVECs and ginkgolide B recovered the phenome-non in glucose-treated endothelial cells.The level of ROS generation was increased in high glucose-treated group,whereas ginkgolide B inhibited ROS genera-tion.Immunofluorescence data showed high glucose in-creased apoptosis,whereas ginkgolide B inhibited ap-optosis in high glucose-treated HUVECs.Moreover, the expressions of Bax and caspase-3 were increased and Bcl-2 was reduced in high glucose-treated group. In contrast,ginkgolide B abolished the expressions of Bax and caspase-3 and increased Bcl-2 expression. Moreover,high glucose enhanced the expression and phosphorylation of p53,while ginkgolide B suppressed the expression and phosphorylation of p53 induced by highglucose.Conclusions GinkgolideBcaninhibit apoptosis and improve transmigration function in high glucose-treated HUVECs.Ginkgolide B has protection against high glucose-induced endothelial cell injury.