细胞内活性氧在甲基乙二醛促人腹膜间皮细胞分泌血管内皮生长因子中的作用  

Effect of glucose degradation products on the expression of VEGF and the regulation of reactive oxygen species in human peritoneal mesothelial cells

作　　者：洪富源[1] 孙芳[2] 刘军[2] 姚建[2] 黄一新[2] 唐知还[2] 

机构地区：[1]复旦大学 [2]上海市第一人民医院肾内科,上海200080

出　　处：《肾脏病与透析肾移植杂志》2005年第4期329-332,376,共5页Chinese Journal of Nephrology,Dialysis & Transplantation

摘　　要：目的:观察葡萄糖降解产物甲基乙二醛(MGO)对人腹膜间皮细胞(HPMC)分泌血管内皮生长因子(VEGF)的影响及细胞内活性氧(ROS)在其中的作用。方法:分别用不同浓度的MGO及抗氧化剂N-酰-L-半胱氨酸(NAC)作用于细胞,用RT-PCR和ELISA方法测定HPMC中VEGF的表达;再以氧化敏感的荧光染料2、7-二氢二氯荧光素(DCFH)染色,流式细胞仪测定细胞内ROS强度。结果:MGO能使细胞内ROS水平明显升高,呈浓度依赖效应;MGO同时以时效和量效方式促进HPMC中VEGF的表达;而NAC能够明显抑制MGO导致的细胞内ROS升高,同时抑制HPMC中VEGF的分泌。结论:MGO可能部分通过诱导细胞内ROS,促进HPMC表达VEGF,从而引起腹膜新生血管的增加,最终导致腹膜失超滤现象。Objective： To study the effects of glucose degradation products （methylglyoxal, MGO） on the production of VEGF in human peritoneal mesothelial cells（HPMC） in vitro and the role of reactive oxygen species （ROS） in this course. Methodology： The MGO （0, 10, 20, 40, 80μM） with or without N-acetyl-L-cysteine （NAC 30 mM） was used in culture medium to stimulate the HPMC. The mRNA level of VEGF was measured with semi-quantitative RT-PCR, more-over, the protein level of VEGF in HPMC was detected by ELISA. The cells were marked with oxidation-susceptible fiuroscent probe 2,7-dichorofiuoresin diacetate （DCFH） and were assayed by flow cytometry. Results： MGO increased the concentration of ROS in the HPMC with a dose-dependent manner. MGO also stimulated the production of VEGF in a dose and time dependent way; however, the effects of MGO on HPMC could be ameliorated by antioxidant NAC. Conclusion： The induction of ROS in HPMC by MGO, which stimulates the expression of VEGF. It may play an important role in the anglogenesis of peritoneum and leads to ultrafihration failure.

关 键 词：人腹膜间皮细胞 活性氧 血管内皮生长因子 腹膜透析 分泌血管内皮生长因子 细胞内活性氧 甲基乙二醛 血管内皮生长因子(VEGF) 细胞内ROS ELISA方法 

分 类 号：R363[医药卫生—病理学]