SIRT1及其相关信号通路在白藜芦醇抑制血管内皮 - 第三军医大学学报.DOC

SIRT1/UCP2通路在白藜芦醇抑制血管内皮细胞氧化应激损伤中的作用 周 曦，易 龙，金 鑫，陈明亮，陈春烨，王 丽，高燕翔，施琳颖，糜漫天 （400038重庆，第三军医大学军事预防医学院营养与食品安全研究中心，重庆市营养与食品安全重点实验室，重庆市医学营养研究中心） [摘要] 目的 观察白藜芦醇对血管内皮细胞氧化应激损伤的抑制作用，并围绕SIRT1/UCP2信号通路探讨其作用机制。方法 原代培养人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs），用叔丁基过氧化氢（t-BHP）而Sirt1抑制剂尼克酰胺能削弱白藜芦醇的抑制作用。结论 白藜芦醇可能通过活化SIRT1抑制UCP2表达，削弱t-BHP诱导的细胞内ROS生成，从而抑制血管内皮细胞氧化应激损伤。 [关键词] 动脉粥样硬化；白藜芦醇；人脐静脉内皮细胞；氧化应激；SIRT1；UCP2 [中图法分类号] [文献标志码] A Role of SIRT1/UCP2 signalling pathway in the inhibition of oxidative injury in the vascular endothelial cells by resveratrol Zhou Xi,Yi Long, Jin Xin,Chen Mingliang,Chen Chunye,Wang Li,Gao Yanxiang, Shi Linyin, Mi Mantian (Research Center for Nutrition and Food Safety, Chongqing Key laboratory of Nutrition and Food Safety, Institute of Military Preventive Medicine, Third Military Medical University, Chongqing, 400038,China) [Abstract] Objective To assess the role of SIRT1/UCP2 signalling pathway in the inhibition of oxidative damage in vascular endothelial cells by resveratrol. Methods Human umbilical vein endothelial cells were cultured. Cells were treated by t-BHP with different concentrations. The cell viability was measured by CCK-8 assay. The intracellular ROS level was observed by fluorospectrophotometer assay . The mRNA expression of SIRT1 and UCP2 were determined by qRT-PCR assay. The protein expression of SIRT1,UCP2 and Caspase3 were determined by western blot assay . Results Cell viability was significantly（P0.05）inhibited by t-BHP treatment and the IC50 was 80 μmol/L. Pretreatment with different doses (0.1,1,10μmol/L) of resveratrol significantly（P0.05）inhibited the decrease of cell viability and increased intracellular ROS level induced by t-BHP Furthermore, the mRNA and protein expression of SIRT1 were upregulated by resveratrol pretreatment, and that of UCP2 was downregulatedyet abolished by niacinamide pretreatement. Conclusion Resveratrol may inhibit t-BHP-induced endothelial injury t