银杏内酯B抑制高糖诱导内皮细胞凋亡及机制研究  

Ginkgolide B inhibits apoptosis in high glucose-stimulated human umbilical vein endothelial cells

作　　者：陈坤[1] 张明[1] 陈北冬[1] 赵艳阳[1] 吴伟[1] 齐若梅[1] 

机构地区：[1]北京医院/北京老年医学研究所国家老年医学中心卫生部老年医学重点实验室,北京100730

出　　处：《中国药理学通报》2017年第3期378-383,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81471051;81270379;81070231)

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.

关 键 词：银杏内酯B 高糖 人脐静脉内皮细胞 ROS 细胞凋亡 p53 

分 类 号：R284.1[医药卫生—中药学] R322.123[医药卫生—中医学]