【摘要】 目的：探讨多酚类化合物3,5-二羟基-4-甲氧基苯甲醇（DHMBA）对人脐静脉内皮细胞（EA. hy926细胞）缺氧/复氧（H/R）损伤的作用及其机制。方法：采用酸性缺氧液结合厌氧工作站构建H/R模型。将EA. hy926细胞分为对照组、H/R组、H/R+不同剂量DHMBA组、H/R+抗氧化剂依达拉奉组、H/R+活性氧（ROS）促生成剂寡霉素A+DHMBA组。采用CCK-8法检测细胞活力；ELISA法检测细胞内肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)水平；Western blot法检测内皮型一氧化氮合酶（eNOS）和核因子κB(NF-κB) p65蛋白的磷酸化水平；激光共聚焦显微镜检测细胞内一氧化氮（NO）含量；二硝基苯甲酸显色法测定谷胱甘肽（GSH）氧化还原平衡；流式细胞术检测细胞内总ROS水平；硝基四紫唑显色法测定细胞乳酸脱氢酶（LDH）漏出；划痕实验检测细胞的迁移能力。结果：DHMBA在125～1 000μmol/L的浓度下无明显细胞毒性。在H/R刺激的EA. hy926细胞中，DHMBA可以提高细胞活力，抑制NF-κB磷酸化，减少炎症因子TNF-α和IL-6含量，增加eNOS蛋白磷酸化水平及NO含量；DHMBA能通过抑制H/R诱导的EA. hy926细胞ROS超载，恢复细胞内GSH和其氧化态（GSSH）的比例，减少LDH漏出，提高细胞迁移能力。结论：DHMBA可以减轻H/R所致的人脐静脉内皮细胞氧化应激、炎症反应、细胞损伤和功能障碍，这与其降低细胞内ROS水平有关。更多还原

【Abstract】 AIM: To explore the effect of polyphenolic compound 3,5-dihydroxy-4-methoxybenzyl alcohol(DHMBA) on hypoxia/reoxygenation(H/R) injury of human umbilical vein endothelial cells(EA. hy926 cells) and its potential mechanisms. METHODS: To construct an H/R model, the EA. hy926 cells were cultured in an acidic hypoxia buffer while in an anaerobic workstation. The cells were divided into control, H/R, H/R+different doses of DHMBA, H/R+edaravone(antioxidant) and H/R+reactive oxygen species(ROS) inducer oligomycin A+DHMBA groups. Cell viability was measured by CCK-8 assay, and tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) levels in cells were measured by ELISA. Phosphorylation of endothelial nitric oxide synthase(eNOS) and nuclear factor-κB(NF-κB) p65 were measured by Western blot. Intracellular NO levels were determined by laser confocal microscopy. Glutathione(GSH)/glutathione disulfide(GSSG) oxidation balance was determined by the dinitrobenzoic acid chromogenic method. Intracellular ROS levels were measured by flow cytometry. Lactate dehydrogenase(LDH) leakage was determined using nitro blue tetrazolium staining. Scratch assays were performed to assess cell migration. RESULTS: DHMBA exhibited no significant cytotoxicity between 125 and 1 000 μmol/L. In H/R-injured human umbilical vein endothelial cells, DHMBA improved cell survival, inhibited phosphorylation of NF-κB p65, reduced the content of TNF-α and IL-6, and increased phosphorylation of eNOS and NO levels. DHMBA also suppressed ROS overload and restored the ratio between GSH and oxidized GSH, decreased in LDH leakage and increased cell migration in H/R-injured human umbilical vein endothelial cells.CONCLUSION: DHMBA can alleviate H/R-induced oxidative stress, inflammation, cellular damage, and dysfunction, which are associated with the ability of DHMBA to inhibit ROS production in human umbilical vein endothelial cells.更多还原