表没食子儿茶素没食子酸酯改善Nε-羧甲基赖氨酸诱导的牛主动脉内皮细胞高通透性  

EPIGALLOCATECHIN-3-GALLATE AMELIORATES BOVINE AORTIC ENDOTHELIAL CELLS HYPERPERMEABILITY INDUCED BY Nε-CARBOXYMETHYLLYSINE

作　　者：燕乔乔 王小宁 许芳溢 衣卫杰 应晨江[3] 叶晓蕾 YAN Qiao-qiao;WANG Xiao-ning;XU Fang-yi;YI Wei-jie;YING Chen-jiang;YE Xiao-lei(School of Public Health and Management,Wenzhou Medical University,Wenzhou 32503;1Department of Nutrition,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052;2 School of Public Health,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030;3School of Public Health and Management,Binzhou Medical College,Yantai 264003,China)

机构地区：[1]温州医科大学公共卫生与管理学院,温州325035 [2]郑州大学第一附属医院营养科,郑州450052 [3]华中科技大学同济医学院公共卫生学院,武汉430030 [4]滨州医学院公共卫生与管理学院,烟台264003

出　　处：《营养学报》2018年第5期477-482,共6页Acta Nutrimenta Sinica

基　　金：国家自然科学基金(No.81273060)

摘　　要：目的探讨表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate, EGCG)改善Nε-羧甲基赖氨(Nε-carboxymethyl-L-lysine,CML)诱导的血管内皮细胞高通透性及其机制。方法牛主动脉内皮细胞(bovineaortic endothelial cells,BAECs)经EGCG(10μmol/L)预处理12 h后,用100μg/ml羧甲基赖氨酸结合白蛋白(CML-BSA)刺激24 h(EGCG+CML-BSA组)。同时设定正常对照组(BSA组),CML模型组(CML-BSA组)及EGCG处理组(EGCG+BSA组)。采用Transwell弥散模型,以FITC-Dextran标志物漏出法检测细胞单层通透性;试剂盒检测胞内ROS水平;qPCR法测定VE-Cadherin(vascularendothelialcadherin)mRNA表达水平;Western-bloting法测定RAGE(receptor of advanced glycation end products)、VE-Cadherin和MCP-1(monocyte chemoattractant protein-1)蛋白表达水平。结果 EGCG预处理能改善CML诱导的内皮细胞通透性升高(P<0.05),降低细胞内活性氧水平(P<0.05),增加VE-Cadherin蛋白和mRNA表达水平(P<0.05),降低MCP-1蛋白表达水平(P<0.05)。结论 CML可诱导内皮细胞通透性升高,而EGCG预处理能改善CML诱导的内皮细胞高通透性,其机制与EGCG抑制CML-RAGE轴有关。Objective To investigate the effect of epigallocatechin-3-gallate （EGCG） on the enhancement of vascular endothelial cells permeability induced by Nε-carboxymethyl lysine （CML） and its molecular mechanism. Methods Bovine aortic endothelial cells （BAECs） were pretreated with EGCG （10 μmol/L） for 12 h and stimulated with 100 μg/ml carboxymethyllysine-conjugated bovine serum albumin （CML-BSA） for 24 h （EGCG＋ CML-BSA group）. At the same time, the control group （BSA group）, CML group （CML-BSA group） and EGCG treatment group （EGCG＋BSA group） were set up. Transwell was used to detect endothelial cell monolayer permeability by FITC-dextran leakage. The level of reactive oxygen species （ROS） was detected by assay kit. The mRNA expression of VE-cadherin （vascular endothelial cadherin） was detected by qPCR. The protein expression levels of RAGE （receptor of advanced glycation end products）, VE-cadherin and MCP-1 （monocyte chemoattractant protein-l） were measured by Western-blot. Results EGCG pretreatment could ameliorate the hyperpermeability of endothelial cells induced by CML （P〈0.05）, decrease intracellular ROS （P〈0.05）, increase the protein and mRNA expression of VE-cadherin （P〈0.05）, and decrease the protein expression of MCP-1 （P〈0.05）. Conclusion CML can induce endothelial cells hyperpermeability, while EGCG pretreatment can ameliorate endothelial cells hyperpermeabilityinduced by CML. The mechanism is related to the inhibition of CML-RAGE axis by EGCG.

关 键 词：表没食子儿茶素没食子酸酯 Nε-羧甲基赖氨酸 糖基化终末产物 内皮细胞通透性 氧化应激 

分 类 号：R151.2[医药卫生—营养与食品卫生学]