晚期氧化蛋白产物通过活性氧诱导单核细胞分泌肿瘤坏死因子

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (05): 493-497.

晚期氧化蛋白产物通过活性氧诱导单核细胞分泌肿瘤坏死因子

张志辉, 刘尚喜, 侯凡凡, 田建伟, 王力, 刘志强, 陈瑗

南方医科大学南方医院肾内科, 广东, 广州, 510515

出版日期:2005-05-20 发布日期:2005-05-20
基金资助:
收稿日期:2004-11-25。
基金项目:国家自然科学基金重点项目(30330300);国家自然科学基金面上项目(30370370)
作者简介:张志辉(1963- ),男,1985年毕业于第一军医大学医疗系,在读研究生,主治医师,E-mail:zhangzhihui@126.com
通讯作者:刘尚喜,电话:020-61641114-86103,E-mail:liusx@fimmu.com

Advanced oxidation protein products-induced tumor necrosis factor α secretion in monocytes via reactive oxygen species generation

ZHANG Zhi-hui, LIU Shang-xi, HOU Fan-fan, TIAN Jian-wei, WANG Li, LIU Zhi-qiang, CHEN Yuan

南方医科大学南方医院肾内科, 广东, 广州, 510515

Online:2005-05-20 Published:2005-05-20

摘要/Abstract

摘要： 目的探讨晚期氧化蛋白产物(AOPP)对单核细胞分泌肿瘤坏死因子(TNFα)的影响及可能机制.方法以THP-1和人外周血单核细胞为单核细胞模型,与用次氯酸氧化牛血清白蛋白(BSA)制备的AOPP-BSA共同培养,用ELISA法检测培养上清TNFα的释放量,用VICTOR Wallac1420多标记分析系统检测细胞氧化2,7-二氢二氯荧光素产生的荧光量反映活性氧的产生量:用抗氧化剂吡咯烷二硫代氨基甲酸盐(PDTC),NADPH氧化酶抑制剂apocynin和P38磷酸化抑制剂SB20380预处理细胞,探讨AOPP诱导单核细胞分泌TNFα的可能机制.结果 AOPP-BSA能诱导单核细胞TNFα的分泌和细胞内活性氧(ROS)的产生.用PDTC预处理细胞,可以清除AOPP-BSA诱导产生的ROS,同时抑制TNFα的分泌.用apocynin抑制NADPH氧化酶及用SB20380抑制P38磷酸化均能有效地抑制AOPP-BSA诱导的TNFα分泌.结论 AOPP可能通过激活单核细胞NADPH氧化酶,释放ROS,使P38磷酸化,导致TNFα的分泌.

Abstract: Objective To investigate the effect of advanced oxidation protein products (AOPP) on the secretion of tumor necrosis factor α (TNFα) in monocytes and its possible mechanism.Method Human monocyte cell line THP-1 and peripheral blood monocytes were incubated with AOPP-bovine serum albumin(BSA) prepared by incubation of BSA with hypochlorous acid. TNFα in the supernatant of the culture medium of THP-1 cells was measured by enzyme-linked immunosorbent assay and the production of reactive oxygen species (ROS) evaluated by measuring the fluorescent product from the oxidation of an oxidant-sensitive 2,7-dichlorefluorescin using Wallac 1420 multilabel counter. The intracellular signal was observed by pre-treatment of the cells with antioxidant pyrrolidine dithiocarbamate, NADPH oxidase inhibitor apocynin or p38 phosphorylation inhibitor SB203580.Results AOPP-BSA induced TNF-α secretion and ROS production in monocytes. Pretreatment of the cells with pyrrolidine dithiocarbamate scavenged most of ROS and almost completely blocked TNF-α secretion induced by AOPP-BSA. Inhibition of NADPH oxidase by apocynin and p38 phosphorylation by SB203580 could both effectively block AOPP-BSA-induced TNF-α secretion.Conclusion AOPP-BSA induced TNF-α secretion in monocytes, and the intracellular signaling involves ROS produced by activated NADPH oxidase and subsequent p38 phosphorylation.

中图分类号:

Q251

张志辉, 刘尚喜, 侯凡凡, 田建伟, 王力, 刘志强, 陈瑗. 晚期氧化蛋白产物通过活性氧诱导单核细胞分泌肿瘤坏死因子[J]. 南方医科大学学报, 2005, 25(05): 493-497.

ZHANG Zhi-hui, LIU Shang-xi, HOU Fan-fan, TIAN Jian-wei, WANG Li, LIU Zhi-qiang, CHEN Yuan. Advanced oxidation protein products-induced tumor necrosis factor α secretion in monocytes via reactive oxygen species generation[J]. Journal of Southern Medical University, 2005, 25(05): 493-497.

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: 1304-13.
[2] Descamps-Latscha B, Witko-Sarsat V. Oxidative stress in chronic renal failure and hemodialysis[J]. Nephrologie, 2003, 24: 377-9.
[3] Albert AW. Lipid hydroperoxide generation, turnover, and effector action in biological systems[J]. J Lipid Res, 1998, 39(8): 1529-42.
[4] Witko-Sarsat V, Friedlander M, Nguyer-Khoa T, 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] Witko-Sarsat V, Gausson V, Descamps-Latscha B. Are advanced oxidation protein products potential uremic toxins [J] ? Kidney Int Suppl, 2003, 84:S11-4.
[6] Yuo A, Kitagawa S, Motoyoshi K, et al. Rapid priming of human monocytes by human hematopoietic growth factors [J]. Blood,1992, 79: 1553-7.
[7] 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.
[8] Zimmerman J, Herrlinger S, Pruy A, et al. Inflammation enhances cardiovascular risk and mortality in hemodialysis patients [J].Kidney Int, 1999, 55: 648-58.
[9] Panichi V,Miglior M,De Pietro S, et al. C-reactive protein as a marker of chronic inflammation in uremic patients [J]. Blood Purif,2000, 18(3): 183-90.
[10] Stenvinkel P, Heimburger O, Paultre F, et al. Strong association between malnutrition, inflammation, and atherosclerosis in chronic renal failure[J]. Kidney Int, 1999, 55(5): 1899-911.
[11] 梁敏,王力,侯凡凡,等单核细胞活化与终末期肾病炎症反应状态的关系[J].第一军医大学学报,2003,23(8):781-4.Liang M, Wang L, Hou FF, et al. Inflammatory stares in patients with end-stage renal disease: role of monocyte activation [J]. J First Mil Med Univ/Di Yi Jun Yi Da Xue Xue Bao, 2003, 23(8): 781-4.
[12] 陈瑗,周攻,侯凡凡,等.氧化应激与慢性肾功能衰竭透析患者动脉粥样硬化的发生[J].中华肾脏病杂志,2002,18(5):377-8.Chen Y, Zhou M, Hou FF, et al. Oxidative stress and atherogenesis in hemodialysis patients with chronic renal failure [J]. Chin J Nephrol, 2002, 18(5): 377-8.
[13] Schomig M, Eisenhardt A, Ritz E. The microinflammatory state of uremia [J]. Blood Purif, 2000, 18(4): 327-32.
[14] Kalantar-Zadeh K, Stenvinkel P, Pillon L, et al. Inflammation and nutrition in renal insufficiency[J]. Adv Ren Replace Ther, 2003, 10(3): 155-69.
[15] 侯凡凡,张训.慢性肾功能衰竭时晚期糖基化终产物的病理生理[J].中华肾脏病杂志,1997,13(5):312-4.Hou FF, Zhang X. Pathophysiological role of advanced glycation end products in patients with chronic renal failure [J]. Chin J Nephrol,1997, 13(5): 312-4.