7-二氟甲氧基金雀异黄素对H_2O_2诱导血管内皮细胞与单核细胞黏附的抑制作用

目的 探讨7-二氟甲氧基金雀异黄素对氧化应激诱导血管内皮细胞与单核细胞黏附的抑制作用及其机制.方法 荧光分光光度计检测单核细胞与血管内皮细胞的黏附,酶联免疫吸附法检测E选择素和细胞间黏附分子1等黏附分子的释放,Western blotting检测P38丝裂原活化蛋白激酶的磷酸化水平.结果 H2O2处理血管内皮细胞24 h后,内皮细胞-单核细胞的黏附率显著增加,内皮细胞E选择素和细胞间黏附分子1的释放增加;加入7-二氟甲氧基金雀异黄素后,血管内皮细胞-单核细胞的黏附率以及内皮细胞释放E选择素和细胞间黏附分子1等黏附分子呈浓度依赖性减少.H2O2处理血管内皮细胞24 h后,可显著激活P38丝裂原活化蛋白激酶,这一作用可被7-二氟甲氧基金雀异黄素所抑制.给予P38丝裂原活化蛋白激酶特异性抑制荆SB203580亦可阻断H2O2诱导的内皮细胞-单核细胞黏附及内皮细胞E选择素和细胞间黏附分子1的释放.结论 7-二氟甲氧基金雀异黄素对氧化应激诱导的血管内皮细胞与单核细胞黏附具有拮抗作用,其机制可能与其抑制P38丝裂原活化蛋白激酶的激活进而阻断内皮细胞释放黏附分子E选择素和细胞间黏附分子1有关.

Inhibitory Effect of 7-Difluoromethyl-genistein on H_2O_2-induced Adhesion Between Vascular Endothelial Cells and Mononuclear Cells

Aim To investigate the effect of 7-difluoromethyl-genistein(FMGEN) on oxidative stress-induced cell adhesion between vascular endothelial cells and mononuclear cells and the underlying mechanism.Methods Fluorescent light spectrophotometer was used to detect the cell adhesion between vascular endothelial cells and mononuclear cells.The concentrations of E-selectin and intercellular adhesion molecule(ICAM-1) in the cell culture supernatant were determined by enzyme-linked immunosorbent assay(ELISA).The activation of P38 mitogen-activated protein kinase (P38-MAPK) was analyzed by Western blotting.Results Exposure of vascular endothelial cells to H_2O_2 for 24 h increased the adhesion between vascular endothelial cells and mononuclear cells and the release of E-selectin and ICAM-1 in vascular endothelial cells;however,these effects of H_2O_2 were inhibited by FMGEN in a concentration-dependent manner. Treatment of vascular endothelial cells with H_2O_2 for 24 h resulted in the significant activation of P38-MAPK and the activation of P38 induced by H_2O_2 was inhibited by FMGEN.SB203580,a specific inhibitor of P38-MAPK blocked the adhesion between vascular endothelial cells and mononuclear cells and the release of E-selectin and ICAM-1 in vascular endothelial cells induced by H_2O_2.Conclusion FMGEN antagonizes oxidative stress-induced cell adhesion between vascular endothelial cells and mononuclear cells,which is associated with inhibition of the release of E-selectin and ICAM-1 via down-regulating the activation of P38-MAPK.