高糖对腹膜间皮细胞β-连环蛋白及骨桥蛋白表达的作用  

The expression of -catenin and osteopontin in rat peritoneal mesothelial cells stimulated with high glucose

作　　者：霍爱晶[1] 马健飞[1] 杨丽娜[1] 孙立霞[1] 王力宁[1] 

机构地区：[1]中国医科大学附属第一医院肾内科,沈阳110001

出　　处：《中国血液净化》2012年第1期33-37,共5页Chinese Journal of Blood Purification

摘　　要：目的观察高糖对大鼠腹膜间皮细胞(ratperitoneal mesothelial cell RPMC)中β-连环蛋白(β-Catenin)、骨桥蛋白(Osteopontin,OPN)2者mRNA及蛋白表达的影响。方法胰蛋白酶法行RPMC的原代培养和传代,经鉴定第3代细胞融合至80%时分组。分为:正常对照组;不同浓度葡萄糖(1.5%,2.5%,4.25%)组作用24h;2.5%高糖作用不同时间(8,12,24,34,48h)组,细胞免疫组织化学法检测β-Catenin蛋白在细胞上的表达情况,RealTime-RT-PCR技术检测β-Catenin、OPN、TGF-β1mRNA的表达。ELISA法检测OPN蛋白表达。结果高糖刺激下RPMC的β-catenin表达增加,与0h组比较,在8h即出现(但不具有统计学意义),12h具有统计学意义,24h达到高峰,48h仍存在,组间差异均有统计学意义(P<0.05);高糖可刺激RPMCOPN的表达显著增加,呈剂量、时间依赖性(P<0.05);TGF-β表达增加,在8h表达于较高水平,12h降低,24h达到高峰,48h仍存在,与0h组比较,组间差异均有统计学意义(P<0.01);高糖刺激下,β-Catenin、TGF-β1表达显著增加,呈剂量依赖性,组间差异具有统计学意义(P<0.05)。结论高糖通过上调β-Catenin、OPN的表达,引起腹膜损伤导致腹膜纤维化,β-Catenin、OPN参与了腹膜间皮细胞透析相关性腹膜纤维化的病理过程。Objective To observe the expression of-catenin and osteopontin(OPN) in rat peritoneal mesothelial cells(RPMCs) stimulated with high glucose.Methods RPMCs were isolated from rat colic omentum.RPMCs were incubated in the medium containing higher glucose(2.5%) for 8,12,24,34 or 48 h,or in medium containing 1.5%,2.5% or 4.25% glucose for 24 h.RPMCs cultured in regular medium was used as the control group.-catenin and OPN were detected by immunohistochemistry,real-time RT-PCR and ELISA.TGF--mRNA was measured by real-time RT-PCR.Results The expression of -catenin and OPN were significantly increased in the RPMCs groups stimulated with high glucose as compared with those in the control group.In the RPMCs pre-incubated with glucose,the expression of -catenin and OPN were significantly up-regulated,as compared with those cultured in high glucose medium.Conclusion Higher glucose in medium up-regulates the expression of -catenin and OPN in RPMCs.

关 键 词：高糖 腹膜间皮细胞 Β-连环蛋白 骨桥蛋白 转移生长因子-Β 

分 类 号：R459.5[医药卫生—治疗学]