厄贝沙坦对高糖诱导肾小管上皮细胞转分化中GSK-3β表达的影响  

Effects of irbesartan on expression of glycogen synthase kinase-3β in tubular epithelial-mesenchymal transition induced by high glucose

作　　者：闫喆[1,2] 姚芳[1] 史永红[1] 郝军[1] 傅淑霞[2] 李绍梅[2] 段惠军[1] 

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

出　　处：《中国药理学通报》2009年第2期225-229,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：目的探讨糖原合成酶激酶-3β(glycogen synthase ki-nase-3β,GSK-3β)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Ⅱ受体1拮抗剂(AT1Ra)厄贝沙坦对其活性的影响。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磷酸化GSK-3β(p-GSK-3β)在肾小管细胞表达情况;Western blot检测GSK-3β、p-GSK-3β、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(RT-PCR)检测GSK-3β和TGF-β1mRNA表达。结果高糖组较正常糖及甘露醇对照组p-GSK-3β、α-SMA蛋白及TGF-β1mRNA表达增高,E-cadherin表达降低。高糖刺激细胞12hp-GSK-3β表达增强,24h达到高峰。厄贝沙坦能够下调高糖诱导的p-GSK-3β、α-SMA蛋白及TGF-β1mRNA表达,同时逆转E-cadherin下降水平。结论GSK-3β可能参与了高糖诱导的肾小管上皮细胞转分化过程,厄贝沙坦抑制该过程可能是通过调节GSK-3β的活性而实现的。Aim To investigate the effects of irbesartan on the expression of glycogen synthase kinase-3β （GSK-3β） in tubular epithelial-mesenehymal transition（EMT） in HKC induced by high-glucose. Methods human kidney proximal tubular epithelial cell line （HKCs） cultured in vitro were divided into four groups at random ： normal-glucose group, mannitol control group, high-glucose group and high-glucose plus irbesartan group. Immunocytochemistry staining was used to observe expression of phospho-GSK-3β （ p-GSK-3β） ; The protein expressions of GSK-3β, p-GSK-3β, E-cadherin,α-SMA were examined by Western blot;GSK-3β and TGF-β1 mRNA were detected by reverse transcription-polymerase chain reaction （RT-PCR ）. Results Compared with normal-glucose and mannitol control group,the expression of p-GSK-3β and α-SMA significantly increased in HKC cells stimulated by high glucose. However, the expression of E-cadherin decreased in high-glucose group. GSK-3β phosphorylation in HKC was in a time-dependent manner, being significant at 12 h and peak occurring at 24 h. The expression of TGF-β1 mRNA was up-regulated with high-glucose stimulation and reduced in irbesartan group. Irbesartan inhibited high expression of p-GSK-3β and α-SMA, and reversed the expression of E-cadherin induced by high glucose. Conclusions These data indicate that GSK-3β is possibly involved in high glucose-induced tubular epithelial-mesenchymal transition. Irbesartan may reverse tubular EMT by regulating activity of GSK- 3β.

关 键 词：糖原合成酶激酶-3Β E-钙粘蛋白 Α-平滑肌肌动蛋白 上皮细胞转分化 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学] R329.2[医药卫生—基础医学]