糖化终末产物对人腹膜间皮细胞氧化应激的影响

Effects of advanced glycation end products on oxidative stress of human peritoneal mesothelial cells

目的研究不同浓度糖化终末产物（AGEs）对人腹膜间皮细胞（HPMC）氧化应激的影响。方法通过胰酶消化法从非。肾脏疾病腹腔手术患者的大网膜中得到HPMC，进行原代培养和传代，免疫组化法对培养的细胞进行鉴定。不同浓度的AGEs干预HPMC一定时间后，采用活性氧（ROS）捕获剂双氢-乙酰乙酸二氯荧光黄（DCFH-DA）孵育细胞，通过流式细胞仪检测细胞内的平均荧光强度测定细胞内ROS水平，观察不同浓度的AGEs对HPMC干预后细胞内ROS的变化。结果AGEs明显抑制HPMC活力（P〈0．05），随着浓度和时间的增加，抑制率上升。AGEs干预HPMC后，细胞内的ROS水平明显升高。结论AGEs导致细胞发生氧化应激，其对细胞的损伤作用可能与氧自由基生成过多、ROS蓄积有关。

Objective To explore the mechanism of oxidative stress in human peritoneal mesothelial cells （HPMC） induced by advanced glycation end products （AGEs）. Method The primary HPMC were isolated from human greater omenta without renal disease by trypsin-EDTA enzymatic digestion, and then cells were cultured. The HPMC were characterized by immunohistochemical staining. After being intervened with AGEs of different concentrations, cell viability was detected by MTT, 2,7-dichlorofluorescein diacetate （DCFH-DA） was used as a reactive oxygen species （ROS） capture. The mean fluorescent intensity （MFI） of 2, 7-dichlorofluorescein （DCF）, which was the product of cellular oxidation of DCFH-DA, was detected by flow cytometry, and ROS was thus measured. Result Viability of HPMC was inhibited by AGEs of different concentrations （P〈0. 05） in a doseand time dependent fashion, and the effect was apparent by AGEs above 200 mg/L （P〈0. 01）. Intracellular MFI was elevated after HPMC were intervened with AGEs. ROS production was stimulated markedly by AGEs above 100 mg/L （P〈0. 01）. Conclusion AGEs induced oxidative stress in HPMC by overproduction of ROS.