抗体催化水的氧化对大鼠血管平滑肌细胞过氧化损伤作用

Effect of antibody-catalyzed water oxidation on peroxidation damage of vascular smooth muscle cells in rats

目的通过观察抗体催化水的氧化作用对大鼠血管平滑肌细胞(VSMCs)的过氧化损伤,探讨其在动脉粥样硬化致病过程中的作用和机制。方法体外分离培养大鼠VSMCs,随机分为4组:空白组、中性粒细胞(Neu)+IgG组、Neu+PMA组、Neu+PMA+IgG组。以佛波酯(PMA)激活大鼠中性粒细胞产生单线态氧(^(1)O_(2)),促进抗体发挥催化水的氧化作用;采用台盼蓝法检测VSMCs的死亡率,CCK-8法检测VSMCs的增殖能力,实时荧光定量PCR及免疫组化法检测细胞内增殖细胞核抗原(PCNA)的表达水平,丙二醛(MDA)法检测细胞内脂质过氧化产物。结果VSMCs在培养过程中,当与中性粒细胞和PMA共同孵育时,细胞的死亡率、增殖能力、细胞内PCNA的表达水平及胞内脂质过氧化产物均显著增高,但增加大鼠IgG共同孵育后,其增高水平得到更进一步增强。结论抗体催化水的氧化可能参与了血管平滑肌细胞的过氧化损伤作用。

Objective To investigate the role and mechanism of antibody-catalyzed water oxidation in the pathogenesis of atherosclerosis by observing its peroxidation damage of vascular smooth muscle cells(VSMCs)in rats.Methods The rat VSMCs were isolated and cultured in vitro,and the rat neutrophils were activated by phorbol ester(PMA)to produce singlet oxygen(^(1)O_(2))promoting antibody to play a catalytic role in the oxidation of wate.The mortality and proliferation of VSMCs were evaluated via trypan blue and CCK-8,respectively.The intracellular expression of proliferating cell nuclear antigen(PCNA)was measured with quantitative real-time PCR(qRT-PCR)and immunohistochemistry,and the lipid peroxidation product was determined using malondialdehyde(MDA)method,respectively.Results When VSMCs were incubated with neutrophils and PMA,the mortality and proliferation of VSMCs,the cell mortality,proliferation ability,intracellular PCNA expression level and lipid peroxidation products were significantly increased,and its level of increase was further enhanced by the co-incubation of IgG.Conclusion The antibody-catalyzed water oxidation may be involved in the oxidative damage of vascular smooth muscle cells.