活性氧与高糖对人腹膜间皮细胞增殖的影响  

Reactive oxygen species and high glucose induce growth inhihition of human peritoneal mesothelial cells through arresting cell cycle progression

作　　者：唐知还[1] 姚建[1] 刘军[1] 张树俭[1] 谢匡成[2] 徐琴君[1] 

机构地区：[1]上海市第一人民医院肾内科,200080 [2]上海市第一人民医院中心实验室,200080

出　　处：《中华肾脏病杂志》2003年第4期223-227,共5页Chinese Journal of Nephrology

基　　金：上海市卫生局青年课题基金(131014Y4)

摘　　要：目的 研究不同浓度葡萄糖、活性氧(过氧化氢)、抗氧化剂对人腹膜间皮细胞(HPMC)细胞增殖的影响及其机制。方法 用~3H-胸腺嘧啶(TdR)掺入法测定细胞增殖:用流式细胞仪检测细胞周期的改变;用半定量RT-PCR检测细胞周期调控蛋白p27^(Kip1)mRNA水平;用细胞免疫组化方法和蛋白印迹(Western blotting)的方法检测p27^(Kip1)蛋白水平。结果 高糖及较大剂量的过氧化氢(0.5 mmol/L)均可以抑制HPMC增殖,而细胞周期分析提示细胞停滞于G_1期。高糖加过氧化氢,则增加了后者的毒性作用。高糖及外源性过氧化氢均可以增加p27^(Kip1)蛋白表达。高糖加抗氧化剂,可以改善细胞周期停滞、增殖抑制作用,但p27^(Kip1)的表达未见明显改变。结论 高糖及外源性过氧化氢均可通过增加p27^(Kip1)的表达使细胞周期发生停滞,从而抑制增殖。而高糖的增殖抑制作用还与内源性的活性氧有关,提示临床上抗氧化治疗可能有益。Objective To investigate the effects of high concentration glucose(HG), exogenous hydrogen peroxide and antioxidants on the cell proliferation of human peritoneal mesothelial cells (HPMCs) and its mechanism. Methods All tests were performed in the cultured HPMCs (HMrSV5) in vitro with different concentrations of glucose(0. 1% , 1. 35% ,3. 86% ), hydrogen peroxide(0. 5 mmol/L,0. 1 mmol/L) and antioxidants (pyruvate, catalase) The DNA synthesis was detected by 3H-thymidine incorporation to indicate the cell proliferation. Flow cytometry was used to measure the proportion of G1 phase in the cell cycle. Gene expression of p27Klp1(one of the important cyclin-dependent kinase inhibitor) was examined with semiquantitative reverse-transcription polymerase chain reaction (RT-PCR). Protein expression of p27Klp1 was measured by immunocytochemistry and Western blotting. Results (1) High glucose and small dose hydrogen peroxide (0. 5 mmol/L) inhibited the proliferation of HPMCs. (2) High glucose and exogenous small dose hydrogen peroxide resulted in the G1 phase arrest of HPMC, and the proportion of G1 phase was increased. Besides, the HG enhanced the toxicity effects of exogenous hydrogen peroxide above-mentioned. (3) High glucose, exogenous hydrogen peroxide increased the p27Kip1 protein expression. Adding the antioxidant to high glucose medium, both the arrest of G1 phase and the inhibition of cell proliferation situation were improved, but the p27Kip1 expression did not change significantly. Conclusions Both the HG and exogenous hydrogen peroxide can induce growth inhibition of HPMC through arresting cell cycle progression, which is partly by the increasing expression of the p27Kip1. The effects of the HG are associated with the endogenous reactive oxygen species (ROS). Therefore, it may be helpful to use the antioxidant in the peritoneal dialysis.

关 键 词：活性氧 高糖 腹膜间皮细胞增殖 腹膜透析 葡萄糖 

分 类 号：R692.9[医药卫生—泌尿科学]