氧化应激在腹膜纤维化大鼠模型腹膜间皮细胞转分化中的作用(英文)  

Epithelial-to-mesenchymal transdifferentiation of peritoneal mesothelial cells mediated by oxidative stress in peritoneal fibrosis rats

作　　者：段绍斌[1] 于洁[2] 刘庆[1] 王予慧[1] 潘鹏[1] 肖力[1] 凌光辉[1] 刘伏友[1] 

机构地区：[1]中南大学湘雅二医院肾内科,中南大学肾病研究所,肾脏疾病与血液净化学湖南省重点实验室,长沙410011 [2]河南省安阳市人民医院,河南安阳455000

出　　处：《中南大学学报（医学版）》2011年第1期34-43,共10页Journal of Central South University ：Medical Science

基　　金：supported by the Department of Science and Technology of Hunan Province,P.R.China(2008JT3005)

摘　　要：目的:研究氧化应激在腹膜纤维化大鼠腹膜间皮细胞转分化中的作用;观察抗氧化剂普罗布考的干预效应。方法:采用4.25%高糖腹透液以100mL/kg腹腔注射法复制腹膜纤维化大鼠模型,实验分为正常对照组、生理盐水对照组、腹膜纤维化模型组和普罗布考干预组。4周后行腹膜平衡试验,处死大鼠,准确测量超滤量,检测腹膜功能;取肠系膜组织匀浆后测定氧化应激指标丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)含量;留取壁层腹膜组织行HE及Masson染色,并测量腹膜厚度;采用免疫组织化学和Western印迹检测大鼠壁层腹膜组织腹膜间皮细胞转分化指标E-cadherin蛋白和α-平滑肌肌动蛋白(SMA)的表达。结果:腹膜纤维化模型组大鼠腹膜间皮细胞脱落,与正常对照组比较,壁层腹膜厚度和肠系膜匀浆MDA含量增加(P<0.01),腹腔超滤量和GSH-Px活性降低(P<0.01);转分化指标α-SMA蛋白表达明显上调(P<0.01),E-cadherin蛋白显著下调(P<0.01)。普罗布考干预组间皮细胞基本完好,与纤维化模型组比较,壁层腹膜厚度和肠系膜匀浆MDA含量降低(P<0.01),腹腔超滤量(P<0.05)和GSH-Px活性(P<0.01)上升;腹膜α-SMA蛋白表达明显下调(P<0.01),E-cad-herin蛋白表达明显上调(P<0.01)。结论:氧化应激在高糖腹透液诱导的大鼠腹膜间皮细胞转分化中起一定作用;普罗布考可改善腹膜纤维化大鼠模型腹膜结构和功能,部分逆转腹膜间皮细胞转分化。Objective To investigate the role of oxidative stress in the epithelial-to-mesenchymal transdifferentiation（EMT） of peritoneal mesothelial cells in rat model of peritoneal fibrosis and the effect of probucol on peritoneal fibrosis.Methods The rat model of peritoneal fibrosis was induced by 4.25% high glucose peritoneal dialysis fluid（PDF）.The rats were randomly divided into 4 groups：the control group,the saline group,the peritoneal fibrosis group,and the probucol group.A 4 hour peritoneal equilibration test（PET） was performed 4 weeks later.The peritoneal function and net ultrafiltration（UF） volume were determined.The level of malondialdehyde（MDA） and glutathione peroxidase（GSH-Px） in peritoneal tissue were examined.The histology of peritoneal membrane was evaluated by light microscopy.E-cadherin and α-smooth muscle actin（α-SMA） protein expression was evaluated by immunohistochemical method and Western blot.Results The mesothelial cells were detached from peritoneal membrane in peritoneal firbosis rats.Comparing with the control rats,the thickness of visceral peritoneum,the level of MDA,and the-SMA protein expression were increased while the net ultrafiltration volume,the level of GSH-Px and E-cadherin protein expression were decreased in peritoneal firbosis rats.All these changes were reversed in the rats treated with probucol.Conclusion Oxidative stress plays an important role in transdifferentiation of peritoneal mesothelial cell in the peritoneal fibrosis rats.Probucol can improve structure and function of peritoneum,and partially reverse the EMT by reducing the oxidative stress.

关 键 词：腹膜透析 腹膜纤维化 腹膜间皮细胞 转分化 氧化应激 普罗布考 

分 类 号：R965[医药卫生—药理学]