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  

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

作　　者：张志辉[1] 刘尚喜[1] 侯凡凡[1] 田建伟[1] 王力[1] 刘志强[1] 陈瑗[1] 

机构地区：[1]南方医科大学南方医院肾内科,广东广州510515

出　　处：《第一军医大学学报》2005年第5期493-497,共5页Journal of First Military Medical University

基　　金：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)~~

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VICTORWallac1420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SB203580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SB203580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。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.

关 键 词：晚期氧化蛋白产物 单核细胞 肿瘤坏死因子 NADPH氧化酶 P38 

分 类 号：R692.5[医药卫生—泌尿科学]