细胞内活性氧在糖基化终末产物促人腹膜间皮细胞分泌单核细胞趋化因子1中的作用

The role of reactive oxygen species in the promotion of monocyte chemoattractant protein-1(MCP-1) production by advanced glycation end products-human serum albumin(AGE-HSA) in human peritoneal mesothelial cells

目的观察糖基化终末产物(advanced glycation end products,AGEs)对人腹膜间皮细胞(humanperitoneal mesothelial cell,HPMC)分泌单核细胞趋化蛋白1(monocyte chemoattractant protein-1,MCP-1)的作用及细胞内活性氧族(reactive oxygen species,ROS)在其中的作用。方法分别用不同浓度的糖基化人血清白蛋白(advanced glycation end product s-human serum albumin,AGE HSA)及抗氧化剂N乙酰-L半胱氨酸(N acetyl-L cysteine,NAC)作用于细胞,用逆转录多聚酶链反应(RT PCR)法和酶联免疫吸附法(E L I S A)测定HPMC中MC P 1的表达;再以氧化敏感的荧光染料2,7-二氢二氯荧光素(2,7-dichlorofluorescin,DCFH)染色,流式细胞仪测定ROS强度。结果 AGE HSA能使细胞内ROS水平明显升高,呈现浓度依赖效应;AGE HSA同时以时效和量效方式促进HPMC中MCP-1的表达;而NAC能够明显抑制AGE-HSA所导致的细胞内ROS升高,同时抑制HPMC中MCP-1的分泌。结论 AGE HSA可能部分通过诱导细胞内ROS,促进HPMC表达MCP-1。

Objective To study the effects of advanced glycation end products-human serum albumin （AGE- HSA） on the production of monocyte chemoattractant protein- 1 （MCP- 1） in human peritoneal mesothelial cells （HPMC） and the role of reactive oxygen species （ROS） in this process. Methods AGE-HSA （0, 100, 500 and 1000μg/ mL） with or without 30mM N-acetyl-L-cysteine （NAC） were added to the cell culture medium to stimulate HPMC. MCP-1 mRNA was measured by semi-quantitative RT-PCR, and MCP-1 protein in HPMC was determined by ELISA. The cells were marked with oxidation-susceptible flurorescent probe 2,7-dichorofluoresin diacetate （DCFH） and then were assayed by flow cytometry. Results AGE-HSA increased the concentration of ROS in HPMC with a dose-dependent manner. AGE-HSA also stimulated the production of MCP-1 in dose- and time- dependent manners. The effects of AGE-HSA on HPMC could be blocked by antioxidant NAC. Conclusion The induction of ROS by AGE-HSA in HPMC stimulates the expression of MCP-1. This process may play an important role in the inflammatory reaction and may lead to the ultrafiltration failure.