3, 5-二羟基-4-甲氧基苯甲醇减轻人脐静脉内皮 细胞缺氧/复氧损伤的作用及其机制

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. [ABSTRACT FROM AUTHOR]