卡维地洛对过氧化氢致血管内皮细胞氧化应激损伤的保护作用

Protective effects of carvedilol on oxidative stress injury induced by hydrogen peroxide in vascular endothelial cells

目的观察卡维地洛(carved ilol)对过氧化氢(hydrogenperoxide,H2O2)致内皮细胞损伤及表面粘附分子表达的影响。方法采用H2O2作为外源性自由基生成系统,模拟内皮细胞的脂质过氧化损伤,建立离体培养的ECV-304细胞氧化应激损伤模型,观察卡维地洛对H2O2致内皮细胞损伤及表面粘附分子表达的影响。结果卡维地洛各浓度组均明显改善H2O2(1.0×10-6mol.L-1)所致ECV-304细胞形态学损伤,提高细胞生存率,降低LDH释放,并可使细胞内及细胞培养液中MDA含量降低,SOD活性升高,亦可下调ICAM-1蛋白及细胞内ICAM-1mRNA表达水平,上述作用随药物浓度增加呈增强趋势。结论卡维地洛可保护内皮细胞结构和功能的完整性,提高内皮细胞抗氧化能力,并从转录水平抑制脂质过氧化诱导的粘附分子表达增加,降低单核-内皮细胞粘附,有利于减少动脉粥样硬化的始动环节和早期事件的发生。

Aim To observe the influence of carvedilol on the injury and expression of intercellular adhesion molecule-1 induced by hydrogen peroxide in ECV-304 cells and investigate the anti-atherosclerotic effect of carvedilol. Methods： The viability of ECV-304 cells was detected by MTT assay. Morphological changes of ECV-304 cells were observed under converse microscope. The level of lactate dehydrogenase released to the extracellular medium, the intracellular superoxide dismutase activity and the extracellular and intracellular Malondialdelyde level were determined using automatic biochemistry analyser. The expression of ICAM-1 in protein level and mRNA level was detected with flow cytometfic technique and RT-PCR. Results Pretreated with carvidilol （1.0 × 10^-5 ～ 1.0 × 10^-9 mol · L^-1 ） for 24 h, the cell survival rate was increased significantly in a concentration-dependent manner. Preincubation for 24 h with carvedilol results in a significant concentration-dependent decline of LDH release from hydrogen peroxide （ 1.0 × 10 ^-6mol · L^-1 ）injured cells. While ECV-304 cells were pre-incubated with carvedilol, the level of MDA decreased and the activity of SOD increased significantly. Carvedilol produced a concentration-dependent inhibition of the expression of ICAM-1 protein and mRNA in hydrogen peroxide injured ECV-304 cells in a similar manner. Conclusion： These experiments demonstrated that carvedilol was able to protect ECV-304 cells from the oxidative stress injury and inhibit ICAM-lexpression in ECV-304 cells induced by hydrogen peroxide. Therefore, we can consider that carvedilol maintains and improves the function of endothelium damaged by hydrogen peroxide from many aspects, which does indicate extensive antioxidant effects on the hydrogen peroxide-injured vascular endothelial cells and suggest promising effects in atherogenesis process.