摘要
目的:探讨内质网应激是否参与氧化三甲胺(TMAO)促人脐静脉内皮细胞(HUVECs)氧化应激的过程。方法:体外培养HUVECs;CCK-8法测定细胞活力;DCFH-DA染色、倒置相差荧光显微镜观察及流式细胞术检测内皮细胞内活性氧簇(ROS)水平;Western blot法检测内皮细胞中phospho-IRE-1α、IRE-1α和GRP78/BiP的蛋白水平。结果:TMAO未表现出对HUVECs活力有显著影响;作用较长时间(>24 h),较低浓度(10μmol/L)的TMAO即可引起原代HUVECs氧化应激增加(P<0.05);TMAO可引起HUVECs IRE-1α磷酸化水平和GRP78/BiP蛋白水平显著升高(P<0.01);IRE1α特异性抑制剂STF-083010预处理1 h可缓解TMAO促HUVECs氧化应激作用(P<0.05)。结论:内质网应激参与了TMAO致HUVECs氧化应激的过程。
AIM:To investigate whether endoplasmic reticulum stress is involved in trimethylamine N-oxide(TMAO)-mediated oxidative stress in human umbilical vein endothelial cells(HUVECs).METHODS:The cell viability was examined by CCK-8 assay.The cells were stained by DCFH-DA,and the intracellular level of reactive oxygen species(ROS)was observed by phase-contrast microscopy and detected by flow cytometric analysis.The protein levels of phospho-IRE-1α,IRE-1αand GRP78/BiP were detected by Western blot.RESULTS:TMAO exerted no significant effect on the viability of HUVECs.For a long period(>24 h),even a low concentration(10μmol/L)of TMAO increased the oxidative stress level in the HUVECs(P<0.05).TMAO increased the phosphorylation level of IRE-1αand significantly up-regulated the protein level of GRP78/BiP in HUVECs(P<0.01).Pretreatment with STF-083010,an inhibitor of IRE1α,for 1 h reduced TMAO-induced oxidative stress in HUVECs(P<0.05).CONCLUSION:Endoplasmic reticulum stress is involved in TMAO-induced oxidative stress in HUVECs.
作者
温仁辉
柯世业
石光耀
刘定辉
余显冠
凌叶盛
朱洁明
钱孝贤
WEN Ren-hui;KE Shi-ye;SHI Guang-yao;LIU Ding-hui;YU Xian-guan;LING Ye-sheng;ZHU Jie-ming;QIAN Xiao-xian(Department of Cardiology,The Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510630,China;Institute of Integrated Traditional Chinese and Western Medicine,Sun Yat-sen University,Guangzhou 510630,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第2期193-199,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81370447)
关键词
氧化三甲胺
内质网应激
氧化应激
人脐静脉内皮细胞
Trimethylamine N-oxide
Endoplasmic reticulum stress
Oxidative stress
Human umbilical vein endothelial cells
