异丙酚对H_2O_2增强肿瘤坏死因子-α诱导血管内皮细胞凋亡效应的抑制作用

Protective Effect of Propofol Against H_2O_2 Potentiated TNF-alpha Induced Human Vascular Endothelial Cell Apoptosis

目的:探索H2O2对TNF-α诱导血管内皮细胞凋亡的作用及异丙酚(propofol)对细胞凋亡的保护作用和可能的机制。方法:将体外培养的ECV304细胞分为5组:①对照组(Control);②10μmol/L H2O2组(H);③40 nmol/L TNF-α组(T);④TNF-α+H2O2组(T+H);⑤TNF-α+H2O2+propofol组(T+H+P)。观察:①流式细胞仪检测细胞凋亡率;②丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)含量。结果:10μmol/L的H2O2仅造成少量细胞凋亡,MDA含量变化不显著(P>0.05);与TNF-α共同作用后细胞凋亡率显著增加,且高于二者分别作用之和,细胞内MDA含量也明显增加,SOD和GSH-Px含量明显减少,与TNF-α组相比差异具有显著性(P<0.05);加入异丙酚预处理可使TNF-α和H2O2组凋亡率明显降低,同时伴随细胞内MDA含量降低,SOD和GSH-Px含量增加。结论:H2O2能显著增强TNF-α诱导细胞凋亡的效应,异丙酚通过降低细胞氧化损伤的机制,有效逆转H2O2增强TNF-α诱导细胞凋亡的效应。

Objective： To test the hypothesis that oxygen free radicals can potentiate TNF-α cellular toxicity, which might be reserved by propofol in human vascular endothelial cells. Methods： Cultured human ECV304 cells were divided into five groups： untreated （Control）, treated with 10 μmol/L hydrogen peroxide （H2O2 ） and treated with 40 nmol/L TNF-α alone （T） or in the presence of H2O2（T＋H） or inpropofol ＋ H2O2（T＋H＋P）, with each cultured for 24 hours. Cell apoptosis was assessed by flow cytometry. The concentration of malondialdehyde （MDA）, super-oxide dismutase.（SOD） and glutathione peroxidase （GSH-Px） were detected at the same time. Results： H2O2, at 10 μmol/L, did not cause significant lipid peroxidation, but enhanced TNF-α induced cell apoptosis（P〈0.01）. Propofol at 50μmol/L attenuated TNF-α and H2O2 induced cell apoptosis, accompanied with a decrease in MDA, while with increases in SOD and GSH-Px.Conclusion. H2O2, even at trace concentration, may significantly enhance TNF-α induced endothelial cell apoptosis. Propofol exerts protective effects against H2O2 potentiated TNF-α cell toxicity by reduction of oxidative injury.