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目的研究上调整合素连接激酶(integerin-linked kinase,ILK)对肾小管上皮细胞(human kidney tubular cells,HKC)E-钙黏素(E-Cadherin)表达的影响。肾小管上皮细胞转分化(epithelial-mesenchymal transition,EMT)在肾间质纤维化中发挥重要作用。E-Cadherin是维持肾小管上皮细胞极性的关键蛋白。ILK是调控上皮细胞转分化的重要蛋白,其作用机制目前还不清楚。方法本研究中我们将ILK转染到HKC中过表达,然后用RT-PCR、Western blot及免疫荧光方法观察了ILK对E-Cadherin表达及糖原合成激酶-3β(glycogen synthase kinase 3β,GSK-3β)活性的影响。进一步用磷酸化下游底物GSK-3β抑制剂SB415286观察对E-Cadherin表达的影响。结果 ILK过表达可以明显抑制E-Cadherin表达,增加GSK-3β活性。用GSK-3β抑制剂处理后可以下调E-Cadherin的表达。免疫荧光显示ILK过表达可使上皮细胞发生极性改变,E-Cadherin表达下调。表明ILK上调可通过激活GSK抑制E-Cadherin表达,进而诱导肾小管上皮细胞出现转分化表型。
Objective To investigate the effects of up-regulation of the expression of integrin-linked kinase (IL-1) on the expression of E-cadherin in human renal tubular cells (HKC). Epithelial-epithelial-mesenchymal transition (EMT) plays an important role in renal interstitial fibrosis. E-Cadherin is a key protein that maintains the tubular epithelial cell polarity. ILK is an important protein that regulates epithelial cell transdifferentiation, and its mechanism of action is unclear. Methods In this study, we transfected ILK into HKC overexpressing cells. The effects of ILK on the expression of E-Cadherin and glycogen synthase kinase 3β (GSK-3β) were observed by RT-PCR, Western blot and immunofluorescence. 3β) activity. The effect of GSK-3β inhibitor SB415286, a downstream substrate of phosphorylation, on E-Cadherin expression was further observed. Results ILK overexpression significantly inhibited E-Cadherin expression and increased GSK-3β activity. Treatment with GSK-3β inhibitor down-regulates E-Cadherin expression. Immunofluorescence showed that the overexpression of ILK could cause the polarity of epithelial cells to change and the expression of E-Cadherin down-regulated. It shows that upregulation of ILK can inhibit the expression of E-Cadherin by activating GSK, and then induce the transdifferentiation phenotype of renal tubular epithelial cells.