晚期氧化蛋白产物通过活性氧诱导人腹膜间皮细胞TGF-β_1的表达  

Advanced oxidation protein products induce human peritoneal mesothelial cells expressing transforming growth factor-beta 1 via the generation of reactive oxygen species

作　　者：陈吉林[1] 史全明[2] 赵久阳[1] 张怡玲[1] 张丽华[3] 徐茂兰[3] 

机构地区：[1]大连医科大学附属二院肾内科,大连116041 [2]东北电力大学校医院,吉林132012 [3]中国科学院大连化物所,大连116041

出　　处：《中国血液净化》2008年第7期370-374,379,共6页Chinese Journal of Blood Purification

摘　　要：目的探讨晚期氧化蛋白产物(AOPP)诱导体外培养的人腹膜间皮细胞(HPMCs)对转化生长因子(TGF-β1)表达的影响及内源性活性氧(ROS)在此过程中的调节机制。方法体外制备AOPP-HSA模型,原代培养人腹膜间皮细胞。采用ELISA法检测TGF-β1蛋白水平,采用RT-PCR半定量分析HPMCs中TGF-β1mRNA的基因表达水平,同时应用流式细胞仪检测细胞内活性氧的水平。结果AOPP-HSA能显著诱导人腹膜间皮细胞TGF-β1的分泌与基因表达,同时增加细胞内ROS的生成,呈时间与剂量依赖关系(P<0.01),不同浓度的抗氧化剂维生素E和N-乙酰半胱胺酸预处理细胞,可明显地降低细胞内ROS的生成量,同时显著地抑制AOPP-HSA诱导人腹膜间皮细胞TGF-β1的分泌与基因表达,呈剂量依赖关系(P<0.01),其中维生素E(50μmol/L)组和NAC(10mmol/L)组抑制更加显著。结论体外制备的AOPP可显著诱导人腹膜间皮细胞TGF-β1的分泌与基因表达,部分可能通过内源性ROS介导细胞内信号转导途径调节此过程,抗氧化剂维生素E和N-乙酰半胱胺酸可显著降低细胞ROS的生成量和显著抑制TGF-β1的分泌与基因表达。此研究揭示抗氧化剂在防治腹膜纤维化发生过程也许是一种可行的治疗策略。Objective To investigate advanced oxidation protein products （AOPP） on the expression of transforming growth factor-β1 （TGF-β1） in human peritoneal mesothelial cells （HPMCs） in vitro, and the regulatory effect of endogenous reactive oxygen species （ROS） in this process. Method The model of AOPP-human serum albumin （HSA） was prepared in vitro. TGF-β1 in cultured primary HPMCS was measured by ELISA, TGF-β1 mRNA was determined by RT-PCR, and intracellular ROS was assessed by flow cytometry. Result AOPP-HSA induced the secretion and gene expression of TGF-β1 and the increase of intracellular ROS production in cultured HPMCs in a time- and dose-dependent manner （P 〈 0.01）. Pretreatment of HPMCs with antioxidant vitamin E and N-acetyl-cyteine inhibited the AOPP-induced ROS production and the secretion and gene expression of TGF-β1 in the cells in a dosedependent manner （P 〈 0.01）. Cells treated with 50μmol/L vitamin E and 10mmol/L N-acetyl-cyteine exhibited the strongest inhibition. Conclusion AOPP induces the expression of TGF-β1 in HPMCs in vitro, which may be mediated, at least partially, by the ROS-dependent cell signaling pathway. Vitamin E and N-acetyl-cyteine significantly inhibit the secretion and gene expression of TGF-β1 via scavenging ROS in HPMCs. The results of this study may provide a therapeutic strategy for the prevention of peritoneal fibrosis.

关 键 词：晚期氧化蛋白产物 人腹膜间皮细胞 转化生长因子Β1 腹膜透析 

分 类 号：R446.6[医药卫生—诊断学]