晚期氧化蛋白产物诱导内皮细胞产生活性氧

南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (12): 1350-1352.

晚期氧化蛋白产物诱导内皮细胞产生活性氧

袁方, 刘尚喜, 田建伟

南方医科大学南方医院肾内科, 广东, 广州, 510515

出版日期:2004-12-20 发布日期:2004-12-20
基金资助:
收稿日期:2004-5-19。
基金项目:国家自然科学基金(30370370);南方医院归国启动基金;
作者简介:袁方(1972- ),女,1995年毕业于内蒙古医学院,现为南方医科大学在读硕士研究生,主治医师.
通讯作者:刘尚喜,电话:020-61641594,E-mail:liusx@fimmu.com

Advanced oxidation protein products induce reactive oxygen species production in endothelial cells

YUAN Fang, LIU Shang-xi, TIAN Jian-wei

南方医科大学南方医院肾内科, 广东, 广州, 510515

Online:2004-12-20 Published:2004-12-20

摘要/Abstract

摘要： 目的探讨晚期氧化蛋白产物(AOPP)对内皮细胞产生活性氧的影响及机制。方法体外用次氯酸氧化牛血清白蛋白(BSA)制备AOPP-BSA,以内皮细胞株ECV304为内皮细胞模型,用VICTOR1420多标记分析系统动态检测细胞氧化2,7-二氢二氯荧光素(DCFH)产生的荧光量,间接反映活性氧的产生量。结果 AOPP能诱导内皮细胞产生活性氧。活性氧的产生量随BSA氧化程度的升高和AOPP-BSA浓度的增加而增多。用硒谷胱甘肽过氧化物酶小分子模拟物ebselen和NADPH氧化酶抑制剂apocynin预处理细胞,可分别将AOPP-BSA诱导产生的活性氧抑制65%和29%。结论 AOPP能够诱导内皮细胞产生活性氧,其部分机制可能与NADPH氧化酶激活有关。

Abstract: Objective To investigate the effect of advanced oxidation protein products (AOPP) on production of reactive oxygen species (ROS) in endothelial cells. Methods Human umbilical vein endothelial cell line ECV304 were stimulated with AOPP-modified bovine serum albumin (AOPP-BSA) prepared by oxidation of BSA with HOCl in vitro. ROS production in the cells was evaluated by kinetic measurement of dichlorofluoroscein (DCF) fluorescence produced by oxidation of an oxidant-sensitive dye 2,7-dichlorefluorescin (DCFH) using VICTOR 1420 multilabel counter. Results AOPP-BSA could induce ROS production in ECV304 cells time- and dose-dependently, and the quantity of ROS increased with the oxidation degree of BSA. Pretreatment of cells with a small-molecular-mass glutathione peroxidase mimic ebselen or NADPH oxidase inhibitor apocynin inhibited ROS production by 65% and 29% respectively. Conclusion AOPP-BSA induces ROS production in endothelial cells, partially mediated by NADPH oxidase activation.

中图分类号:

Q251

袁方, 刘尚喜, 田建伟. 晚期氧化蛋白产物诱导内皮细胞产生活性氧[J]. 南方医科大学学报, 2004, 24(12): 1350-1352.

YUAN Fang, LIU Shang-xi, TIAN Jian-wei. Advanced oxidation protein products induce reactive oxygen species production in endothelial cells[J]. Journal of Southern Medical University, 2004, 24(12): 1350-1352.

参考文献

[1] Witko-Sarsat V, Friedlander M, Capeillere-Blandin C, et al. Advanced oxidation protein products as a novel marker of oxidative stress in uremia[J]. Kidney Int, 1996,49(5): 1304-13.
[2] Kaneda H, Taguchi J, Ogasawara K, et al. Increased level of advanced oxidation protein products in patients with coronary artery disease[J]. Atherosclerosis, 2002, 162(1): 221-5.
[3] Drueke T, Witko-Sarsat V, Massy Z, et al. Iron therapy, advanced oxidation protein products, and carotid artery intima-media thickness in end-stage renal disease [J]. Circulation, 2002, 106(17): 2212-17.
[4] Witko-Sarsat V, Friedlander M, Nguyen AT, et al. Advanced oxidation protein products as novel mediators of inflammation and monocyte activation in chronic renal failure [J]. J Immunol, 1998,161(5): 2524-32.
[5] Reznick AZ, Packer L. Oxidative damage to proteins: spectrophotometric method for carbonyl assay [J]. Methods Enzymol, 1994,233: 357-63.
[6] Carter WO, Narayanan PK, Robinson JP intracellular hydrogen peroxide and superoxide anion detection in endothelial cells [J]. J Leukoc Biol, 1994, 55(2): 253-8.
[7] Van den Worm E, Beukelman CJ, Van den Berg AJ, et al. Effects of methoxylation of apocynin and analogs on the inhibition of reactive oxygen species production by stimulated human neutrophils [J]. Eur J Pharmacol, 2001, 433(2-3): 225-30.
[8] Liu SX, Chen Y, Zhou M, et al. Inhibitory effect ofebselen on the oxidation of low density lipoprotein[J]. Redox Rep, 1995, 1:357-60.
[9] Witko-SarsatV, GaussonV, NguyenAT, et al. AOPP-induced activation of human neutrophil and monocyte oxidative metabolism:a potential target for N-acetylcysteine treatment in dialysis patients [J]. Kidney Int, 2003, 64(1): 82-91.
[10] Wautier MP, Chappey O, Corda S, et al. Activation of NADPH oxidase by AGE links oxidant stress to altered gene expression via RAGE[J]. Am J Physiol Endocrinol Metab, 2001, 280(5): E685-94.