[1]白志勋,陆静,杨亦彬.TGF-β1/ILK/FSP1信号通路在环孢素诱导肾小管上皮细胞转分化中的作用[J].南方医科大学学报,2019,(07):804.[doi:10.12122/j.issn.1673-4254.2019.07.09]
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TGF-β1/ILK/FSP1信号通路在环孢素诱导肾小管上皮细胞转分化中的作用()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2019年07期
页码:
804
栏目:
出版日期:
2019-07-15

文章信息/Info

Title:
Role of TGF-β1/ILK/FSP1 signaling pathway in cyclosporin A-induced epithelialmesenchymal transition in cultured renal tubular epithelial cells
作者:
白志勋陆静杨亦彬
关键词:
环孢素肾病上皮细胞转分化转化生长因子-β1整合素连接激酶成纤维细胞特异蛋白1
Keywords:
chronic cyclosporine nephropathy epithelial-mesenchymal transition transforming growth factor-β1 integrinlinked kinase fibroblast-specific protein 1
DOI:
10.12122/j.issn.1673-4254.2019.07.09
摘要:
目的通过体外特异阻断试验,探讨转化生长因子-β1/整合素连接激酶/成纤维细胞特异蛋白1(TGF-β1/ILK/FSP1)信号通 路在环孢素A致肾小管上皮细胞转分化中的作用。方法构建并鉴定ILK-RNAi慢病毒表达载体ILKshRNA。将NRK52E细 胞培养分为5组,分别为空白对照组;环孢素诱导组:细胞培养液加环孢素A1 mg/L;TGF-β1干预组:阻断剂(SB431542)10 μmol/L 加环孢素A1 mg/L;ILK干预组:阳性转染ILKshRNA后加入环孢素A1 mg/L;阴性对照组:阴性转染ILKshRNA后加入环孢素 A1 mg/L。实时荧光定量PCR检测TGF-β1、ILK和FSP-1 mRNA的表达,Western blot检测TGF-β1、ILK、FSP-1的蛋白表达。免 疫组化检测细胞爬片α-SMA阳性表达细胞数。结果与空白对照组比较,环孢素诱导组细胞TGF-β1、ILK、FSP-1基因及蛋白表 达量均显著增加(P<0.05);TGF-β1 干预组经SB431542 处理后,TGF-β1、ILK 和FSP1 水平较环孢素诱导组降低(P<0.05);经 ILKshRNA沉默后,ILK和FSP1水平较环孢素诱导组降低(P<0.05)。α-SMA阳性细胞数经SB431542和ILKshRNA处理后较 环孢素诱导组降低(P<0.05)。结论TGF-β1/ILK/FSP1信号通路激活是环孢素A诱导大鼠肾小管上皮细胞转分化的重要机制 之一,ILK参与环孢素A诱导大鼠肾小管上皮细胞间充质转分化过程。
Abstract:
Objective To explore the role of transforming growth factor-β1/integrin- linked kinase/fibroblast-specific protein 1 (TGF- β1/ILK/FSP1) signaling pathway in cyclosporine A (CsA)-induced renal tubular epithelial cell transdifferentiation. Methods Rat renal tubular epithelial NRK-52E cells were induced with 1 mg/L CsA, treated with TGF-β1 inhibitor (SB431542, 10 μmol/L), or transfected with the ILK-RNAi lentiviral expression vector (ILKshRNA) or a negative control vector before CsA induction. The expressions of TGF-β1, ILK and FSP-1 mRNAs and proteins in the cells were detected using real-time PCR and Western blotting. The positive cells for α-SMA expression were detected by immunohistochemistry. Results Compared with the blank control cells, the cells treated with CsA showed significantly increased levels of TGF-β1, ILK and FSP-1 mRNAs and proteins (P<0.05). The expressions of TGF-β1, ILK and FSP-1 were significantly lower in TGF-β1 inhibitor group than in CsA group (P<0.05). The levels of ILK and FSP-1 were significantly decreased after shRNA-mediated ILK silencing (P<0.05). The number of positive cells for α-SMA was significantly lower in cells treated with SB431542 and in cells with ILK silencing than in the cells treated with CsA alone (P<0.05). Conclusion The activation of TGF-β1/ILK/FSP-1 signaling pathway is an important mechanism for CsA-induced transdifferentiation in rat renal tubular epithelial cells. ILK participates in CsA-induced epithelialmesenchymal transition of renal tubular epithelial cells.
更新日期/Last Update: 1900-01-01