【摘要】目的 观察糖基化终末产物(advanced glycation end products,AGEs)对人腹膜间皮细胞(human peritoneal 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 products-human serum albumin,AGE-HAS)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(ELISA)测定HPMC中MCP-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。
【Abstract】 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.
