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几种抗氧化剂对细胞间粘附分子-1表达的影响
目的探讨抗氧化剂调节内皮细胞表面的粘附分子ICAM-1表达的分子生物学机制.方法内皮细胞表面的粘附分子表达用细胞ELISA方法测定,内皮细胞NF-κB的活性用电泳迁移率分析测定.结果 PDTC,DCI和chrysin能明显抑制ICAM-1的表达.Probucol仅在低浓度时对ICAM-1有轻微抑制作用.电泳迁移率分析结果,只有PDTC对NF-κB的活性有抑制作用,其它三种抗氧化药无效.结论这几种抗氧化药抑制内皮细胞表面粘附分子的表达与它们对核转录因子NF-κB的作用并无必然的联系.
关键词： 细胞粘附分子 抗氧化剂 NF-kappa-B -
Objective To investigate whether antioxidants inhibit adhesion of leukocytes to endothelium and furthermore, whether all antioxidants regulate NF-κB activation through a redox sensitive mechanism. Methods The effect of the antioxidative substances pyrrolidin dithiocarbamat (PDTC),dichloroisocumarin (DCI), chrysin and probucol on the endothelial leukocyte adhesion were examined under near physiological flow conditions. The antioxidative activity of antioxidants was measured in a DCF fluorescence assay with flow cytometry. The activation of NF-κB in endothelial cells was investigated in a gel shift assay. Results PDTC and probucol did not show an inhibitory effect to the formation of intracellular H2O2 in TNFct activated human vascular endothelial cells (HUVEC) . Chrysin showed a moderate effect.DCI showed a strong antioxidative effect. In contrast,PDTC and chrysin inhibited the adhesion of HL 60 cells to TNFa-stimulated HUVEC. DCI and probucol did not have influence on the adhesion within the area of the examined shear stresses. Only PDTC inhibited the TNFα-induced activation of NF-kB in endothelial cells.Conclusion The inhibition of the endothelial leukocyte adhesion by antioxidative substances is not to be explained by its antioxidative characteristics only. The inhibitory effect of PDTC on NF-kB activation was probably not related to its antioxidative properties.
