普罗布考和抗氧化维生素不影响体外实验条件下内皮细胞的白细胞粘附

Probucol and Antioxidative Vitamins Showed no Effect on Leukocytes-Endothelial Cells Adhesion in Vitro

为探讨预防动脉粥样硬化的药物普罗布考、维生素C和维生素E是否抑制内皮细胞表面粘附分子表达和白细胞—内皮细胞的粘附 ,以及这种抑制是否通过影响核因子 -κB的活性来实现的 ,在液体流动小室中进行细胞粘附实验 ;用ELISA方法测定内皮细胞粘附分子E -选择素的表达 ;用电泳迁移率分析测定内皮细胞核因子 -κB的活性。经肿瘤坏死因子α刺激的内皮细胞核因子 -κB活性增加 ,粘附分子E -选择素的表达上调 (是基础水平的 3.5倍 ) ,其表面HL6 0细胞的粘附增加 (是基础水平的 4～ 2 6倍 ) ,而抗氧化剂PDTC使所有这些变化都受到抑制。PDTC浓度为 18μmol L时对粘附分子E -选择素的表达呈最大半抑制 ;PDTC浓度为 5 2 μmol L时对内皮细胞表面HL6 0细胞的粘附呈最大半抑制。普罗布考、维生素C和维生素E对肿瘤坏死因子α诱导的粘附分子表达和HL6 0细胞与内皮细胞的粘附没有作用 ;对核因子 -κB的活性没有影响。临床上常用的这三种抗氧化剂并未影响作为动脉粥样硬化始动机制之一的E -选择素介导的白细胞 -内皮细胞粘附水平。

Aim To investigate whether antioxidants probucol, vitamin E and vitamin C modulate expression of endothelial cell adhesion molecules through regulating NF-κB activation. Methods The adhesion of HL60 cell on endothelial cells was measured with adhesion assay in a flow chamber. The effects of the antioxidative substances probucol, vitamin E and vitamin C on the expression of endothelial cell adhesion molecules were measured with cell-ELISA. The activation of NF-κB in endothelial cells was investigated in a gel shift assay. Results In TNFα-activated HUVEC, an increase in p65 and p50, a significantly increased expression of E-selectin (3.5 times), and a significantly increased HL60 cell adhesion to HUVECs (4～26 times) were detected. PDTC inhibited these increases. The half-maximal inhibition of PDTC on E-selectin expression and adhesion of HL60 to HUVEC induced by TNFα was at 18 μmol/L and 52 μmol/L, respectively. Probucol, vitamin E and vitamin C showed no effect on TNFα-induced E-selectin expression in endothelialcells, no influence on HL60-endothelial cell adhesion as well as the TNFα-induced activation of NF-κB in endothelial cells. Conclusions These antioxidants could not inhibit adhesion molecule expression.