【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。更多还原

【Abstract】 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-1α and 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(10 μmol/L) of TMAO increased the oxidative stress level in the HUVECs(P<0.05). TMAO increased the phosphorylation level of IRE-1α and 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.更多还原