转录因子EB在白藜芦醇抑制氧化应激损伤中的内皮保护作用  

Resveratrol protects human umbilical vein endothelial cells against oxidative injury by activating transcription factor EB in vitro

作　　者：周曦[1] 易龙[1] 周敏[1] 张玉 侯鹏飞[1] 刘洋[1] 糜漫天[1] ZHOU Xi;YI Long;ZHOU Min;ZHANG Yu;HOU Pengfei;LIU Yang;MI Mantian(Center of Nutrition and Food Safety,Chongqing Center of Medical Nutrition,Chongqing Key Laboratory of Nutrition and Food Safety,Faculty of Military Preventive Medicine,Army Medical University（Third Military Medical University）,Chongqing,400038,China)

机构地区：[1]陆军军医大学(第三军医大学)军事预防医学系营养与食品安全研究中心,重庆市医学营养研究中心,重庆市营养与食品安全重点实验室,重庆400038

出　　处：《第三军医大学学报》2018年第20期1839-1845,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81502804)

摘　　要：目的观察白藜芦醇(resveratrol,RSV)对血管内皮细胞氧化应激损伤的抑制作用,并探索转录因子EB(transcription factor EB,TFEB)在其中的影响。方法用棕榈酸(palmitate,PA)处理原代人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)建立氧化应激损伤模型。RSV(10μmol/L)预处理2 h,再用PA(200μmol/L)处理24 h,CCK-8法检测细胞增殖活力,荧光酶标仪检测细胞内活性氧(ROS)生成,脂质氧化丙二醛(malondialdehyde,MDA)试剂盒检测细胞脂质过氧化水平。Western blot检测RSV对TFEB蛋白表达的影响。结果 PA(200μmol/L)处理24 h可显著抑制内皮细胞增殖活力,上调胞内ROS水平和MDA水平,导致内皮细胞氧化应激损伤(P <0. 01)。RSV预处理后能显著抑制PA诱导的氧化应激损伤,显著上调TFEB的mRNA和蛋白表达(P <0. 05),而TFEB siRNA处理以后,RSV的保护性作用被明显削弱。结论白藜芦醇是通过激活转录因子EB,从而抑制棕榈酸诱导的内皮细胞氧化应激损伤。Objective To observe the inhibitory effect of resveratrol on oxidative stress in human umbilical vein endothelial cells （HUVECs） in vitro and explore the role of transcription factor EB （TFEB） in mediating the anti-oxidative effect of resveratrol. Methods Primary culture of HUVECs was pretreated with 10 μmol/L resveratrol for 2 h followed by exposure to 200 μmol/L palmitate for 24 h. The proliferation of the cells was assessed using CCK-8 assay, and reactive oxygen species （ROS） and malondialdehyde （MDA） levels in the cells were determined using a commercial detection kit. The mRNA and protein expressions of TFEB were measured using qRT-PCR and Western blotting, respectively. Results Palmitate treatment significantly decreased the viability and increased ROS and MDA levels in cultured HUVECs （P〈0.01）. Pretreatment with resveratrol significantly inhibited PA-induced oxidative injury and up-regulated the mRNA and protein levels of TFEB in the cells; knockdown of TFEB with a specific siRNA obviously attenuated the protective effect of resveratrol against oxidative cell injury （P〈0.05）. Conclusion Resveratrol can protect cultured HUVECs against oxidative injury by activating TFEB.

关 键 词：转录因子EB 白藜芦醇 人脐静脉内皮细胞 氧化应激 动脉粥样硬化 

分 类 号：R282.71[医药卫生—中药学] R285.5[医药卫生—中医学]