厄贝沙坦对高糖诱导肾小管上皮细胞转分化中Wnt/β-catenin信号途径表达的影响  

Modulation of Wnt/β-catenin signaling pathway by irbesartan in highglucose-induced tubular epithelial-mesenchymal transition

作　　者：闫喆[1,2] 姚芳[2] 张丽萍[1] 刘力强[3] 郝军[2] 傅淑霞[1] 段惠军[2] 

机构地区：[1]河北医科大学第二医院肾内科,河北石家庄050000 [2]河北医科大学病理学教研室,河北石家庄050017 [3]河北医科大学第二医院神经外科,河北石家庄050000

出　　处：《中国药理学通报》2009年第12期1630-1634,共5页Chinese Pharmacological Bulletin

基　　金：河北省自然科学基金资助项目(NoC2006000825);河北省卫生厅资助课题(No08282)

摘　　要：目的探讨Wnt/β-catenin信号途径在高糖诱导肾小管上皮细胞转分化中的表达及血管紧张素受体拮抗剂(ARB)厄贝沙坦对该途径活性的影响。方法体外培养人近端肾小管上皮细胞(HKC),分为正常糖组、甘露醇对照组、高糖组、高糖+厄贝沙坦干预组。采用免疫细胞化学观察β-连环蛋白(β-catenin)表达情况;Western印迹检测Wnt4、β-catenin、E-钙粘蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA)表达水平;逆转录-聚合酶链反应检测Wnt4和β-catenin mRNA表达水平。结果高糖组较正常糖及渗透浓度对照组Wnt4蛋白及mRNA、α-SMA蛋白表达增高,E-cadherin表达降低,β-catenin总蛋白及mRNA水平无明显变化,细胞质及核蛋白表达增强。Wnt4和β-catenin核蛋白表达在高糖诱导24h达高峰;厄贝沙坦下调Wnt4、α-SMA及β-catenin核蛋白表达,升高E-cadherin表达水平。结论Wnt/β-catenin信号途径可能参与了高糖介导的肾小管上皮细胞转分化过程。厄贝沙坦可能通过调节Wnt/β-catenin信号途径活性而抑制该过程。Aim To investigate the effects of irbesartan on Wnt/β-catenin signaling pathway in tubular epithelial-mesenchymal transition（EMT）in HKCs induced by high-glucose.Methods Human kidney proximal tubular epithelial cell line（HKCs）cultured in vitro was divided into four groups：normal-glucose group,mannitol control group,high-glucose group and high-glucose plus irbesartan group.Immunocytochemistry staining was used to observe the expression of β-catenin;the protein expression of Wnt4,β-catenin,E-cadherin and α-SMA was assessed by Western blot;Wnt4 and β-catenin mRNA were detected by reverse transcription-polymerase chain reaction（RT-PCR）.Results Compared with normal-glucose and mannitol control group,both the protein and the mRNA of Wnt4 were up-regulated in HKCs stimulated by high-glucose.α-SMA expression significantly increased but E-cadherin decreased in HG group.The cytoplastic and nuclear fraction of β-catenin enhanced with highglucose stimulation.But no difference of the total protein and mRNA of β-catenin was observed between highglucose-treatment and control groups.Highglucose induced Wnt4 and β-catenin expression in a time-dependent manner,both peaking at 24 h.Irbesartan reduced the promotional effect of HG on Wnt4 and α-SMA expression,and nuclear translocation of β-catenin.HG-mediated inhibition of E-cadherin was also restored by irbesartan.Conclusion These data supported a functional role for Wnt/β-catenin signaling pathway during epithelial-mesenchymal transition of HKCs induced by high glucose and suggested that irbesartan might reverse tubular EMT by regulating activity of Wnt/β-catenin pathway.

关 键 词：Wnt4 Β-连环蛋白 E-钙粘蛋白 Α-平滑肌肌动蛋白 上皮细胞转分化 厄贝沙坦 

分 类 号：R-331[医药卫生] R322.61