摘要
目的 研究慢性肾功能衰竭病人循环晚期氧化蛋白产物 (AOPP)对人血管内皮细胞活性的影响及机制。方法 用次氯酸(HOCl)氧化牛血清白蛋白 (BSA)体外制备AOPP BSA ,将人血管内皮细胞株ECV30 4与不同浓度、不同氧化修饰程度的AOPP BSA共同培养 ,用MTT法测定细胞活性 ,用VICTORWallac 14 2 0多标记分析系统动态检测细胞内活性氧的产生量。结果 AOPP BSA使血管内皮细胞活性降低 ,其对内皮细胞活性的影响与BSA氧化程度及AOPP BSA的浓度有关 ;AOPP BSA刺激内皮细胞生成活性氧 ,细胞内活性氧的生成量随BSA氧化程度和AOPP BSA浓度增加而升高。用硒谷胱甘肽过氧化物酶小分子模拟物ebselen预处理细胞 ,可使AOPP BSA诱导产生的活性氧减少 5 3% ,并可保护细胞免受AOPP BSA造成的细胞损伤。结论 晚期氧化蛋白产物能通过氧化应激引起血管内皮细胞损伤。
Objective A close relationship between atherosclerosis and plasma levels of advanced oxidation protein products (AOPP) has been demonstrated in patients with chronic renal failure. The present study was performed to investigate the effect of AOPP on human endothelial cells. Methods Human umbilical vein endothelial cell line ECV304 was incubated with different concentrations of AOPP-BSA, which prepared by combining BSA with HOCl at different molar ratios. Cell viability was measured by MTT assay. Intracellular production of reactive oxygen species (ROS) was evaluated kinetically using VICTOR Wallac 1420 mutilabel counter. Results AOPP-BSA decreased endothelial cell viability, which was dependent on the molar ratio of BSA/HOCl and the concentration of AOPP-BSA. AOPP-BSA also enhanced the intracellular ROS production in ECV304. AOPP-induced ROS production was correlated with the molar ratio of BSA/HOCl and the concentration of AOPP-BSA. Pretreatment of cells with a small molecular glutathione peroxidase mimic (ebselen) reduced AOPP-induced ROS production by 53% with preservation of cell viability. Conclusion AOPP decreased endothelial cell viability via induction of oxidative stress.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2004年第11期951-954,共4页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金重点项目(编号 30 330 30 0 )
广东省团队项目 (编号 1 0 71 7)资助课题
关键词
晚期氧化蛋白产物
内皮
血管
活性氧组分
氧化性应激
advanced oxidation protein products
endothelium, vascular
reactive oxygen species
oxidative stress
