白藜芦醇修复内皮细胞氧化损伤抑制促血栓分子表达的作用和机制  

Resveratrol protects endothelial cells from oxidative damage and inhibits expression of pro-thrombosis molecules:the underlying mechanisms

作　　者：何卫 娄振凯[1] 王兵[1] 李兴国[1] 赵冲宇 赵学凌[1] He Wei;Lou Zhenkai;Wang Bing;Li Xingguo;Zhao Chongyu;Zhao Xueling(Department of Orthopedics,the First Affiliated Hospital of Kunming Medical University,Kunming 650032,Yunnan Province,China)

机构地区：[1]昆明医科大学第一附属医院骨科,云南省昆明市650032

出　　处：《中国组织工程研究》2019年第3期464-469,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81160236);项目负责人:王兵;国家自然科学基金项目(81760029);项目负责人:王兵;国家自然科学基金项目(81760030);云南省卫生科技计划项目(2016NS021;2017NS021;2017NS022);昆明医科大学第一附属医院博士科研基金项目(2017BS030)~~

摘　　要：背景:相关研究表明,白藜芦醇具有抗氧化、抑制动脉血栓的作用。目的:探讨白藜芦醇保护内皮细胞氧化损伤,抑制深静脉血栓形成的分子作用。方法:使用人脐静脉内皮细胞进行传代培养;实验分3组:①对照组为正常培养液中培养24 h;②H_2O_2组在培养液中加入200μmol/L H_2O_2培养24 h;③H_2O_2+白藜芦醇组:先分别予以10,20,30μmol/L的白藜芦醇预处理2 h,再加入200μmol/L H_2O_2培养24 h。采用MTT法检测细胞活力;Hoechst 33258染色法、荧光显微镜观察细胞凋亡;Westernblot检测P-选择素、P-选择素糖蛋白配体1、血管性血友病因子及PI3-K/Akt/GSK3β、NF-κB信号通路关键蛋白表达。结果与结论:①与对照组相比,H_2O_2能够诱导内皮细胞损伤,白藜芦醇对H_2O_2诱导的内皮细胞损伤具有修复作用(P <0.05);②与对照组相比,H_2O_2组细胞凋亡率明显增加(P <0.05);与H_2O_2组相比,H_2O_2+白藜芦醇组细胞凋亡率明显减少(P<0.05);③与对照组相比,H_2O_2组P-选择素、P-选择素糖蛋白配体1、血管性血友病因子、NF-κB和GSK3β蛋白的表达显著上调(P <0.05),Akt总蛋白无明显变化(P> 0.05),pAkt蛋白表达下调(P <0.05);与H_2O_2组相比,H_2O_2+白藜芦醇组上述蛋白表达被逆转(P <0.05);④结果表明,白藜芦醇能够修复静脉内皮细胞氧化损伤,对P-选择素、P-选择素糖蛋白配体1、血管性血友病因子蛋白表达具有抑制作用,其机制可能与PI3-K/Akt/GSK3β、NF-κB信号通路相关。BACKGROUND:Resveratrol has been shown to hold anti-oxidant and anti-arterial thrombosis effects.OBJECTIVE:To explore the mechanism of resveratrol in repairing endothelial cell injury and inhibiting deep vein thrombosis at molecular level.METHODS:Human umbilical vein endothelial cells were isolated and cultured.The cells were divided into three groups:control group(common medium),H2O2 group(cultured with 200μmol/L H2O2 for 24 hours),and resveratrol plus resveratrol group(pretreated with 10,20 and 30μmol/L resveratrol for 2 hours,and then cultured with 200μmol/L H2O2 for 24 hours).The cell viability was evaluated by MTT assay.The apoptotic cells were observed by Hoechst 33258 staining and fluorescence microscopy.The protein expression levels of P-selectin,P-selectin glycoprotein ligand-1,von Willebrand Factor and PI3-K/Akt/GSK3βand NF-κB signaling pathways were tested by western blot assay.RESULTS AND CONCLUTION:Compared with the control group,H2O2 could induce endothelial cell injury and resveratrol could repair endothelial cells injured by H2O2(P<0.05).The apoptotic rate in the H2O2 group was significantly higher than that in the control group(P<0.05).Compared with the H2O2 group,the apoptosis rate in the H2O2 plus resveratrol group was significantly decreased(P<0.05).Compared with the control group,the expression levels of P-selectin,P-selectin glycoprotein ligand-1,von Willebrand Factor,NF-κB and GSK3βin the H2O2 group were significantly increased(P<0.05),the expression of Akt protein showed no significant change(P>0.05),and the expression level of p-Akt protein was significantly down-regulated(P<0.05).Meanwhile,resveratrol could reverse all above protein expression(P<0.05).To conclude,resveratrol can repair human umbilical vein endothelial cells and inhibit the expression of P-selectin,P-selectin glycoprotein ligand-1 and von Willebrand Factor proteins,which may be related to PI3-K/Akt/GSK3βand NF-κB signaling pathways.

关 键 词：白藜芦醇 内皮细胞 氧化应激 国家自然科学基金 组织构建 藜芦属 内皮细胞 氧化性应激 静脉血栓形成 组织工程 

分 类 号：R446[医药卫生—诊断学]