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目的研究转化生长因子β1(TGF-β1)诱导肾小管上皮细胞合成纤连蛋白(FN)与整合素连锁激酶(ILK)表达的关系。方法培养人肾小管上皮细胞(HKC),用蛋白印迹方法检测TGF-β1诱导HKC合成FN和表达ILK的作用。通过基因转染的方法,将含人野生型ILK基因的表达质粒pCMV-wtILK和无激酶活力的人ILK基因表达质粒pCMV-kdILK转染HKC细胞,观察ILK过度表达和抑制ILK活力对TGF-β1诱导的HKC合成FN的影响。结果TGF-β1能够刺激HKC合成FN,其作用呈剂量依赖性。TGF-β1刺激8 h即可上调HKC细胞ILK的表达,与TGF-β1所致的FN合成的增加相一致。转染pCMV-wtILK质粒的HKC细胞,其FN的表达呈增加趋势。转染pCMV-kdILK抑制ILK的活力能够显著拮抗TGF-β1刺激HKC表达FN的作用。结论TGF-B1刺激肾小管上皮细胞FN合成与其所致的ILK表达密切相关。抑制ILK的活力能够拮抗TGF-B1导致的FN合成。
Objective To investigate the relationship between TGF-β1-induced fibronectin (FN) and integrin-linked kinase (ILK) expression in renal tubular epithelial cells. Methods Human renal tubular epithelial cells (HKC) were cultured and the role of TGF-β1 in inducing FN and ILK expression in HKCs was detected by Western blotting. The recombinant plasmid pCMV-wtILK containing human wild-type ILK gene and the human ILK gene expression plasmid pCMV-kdILK without kinase activity were transfected into HKC cells by gene transfection to observe the effect of ILK overexpression and ILK activity on TGF-β1 Effect of induced HKC FN synthesis. Results TGF-β1 stimulated the synthesis of FN by HKC in a dose-dependent manner. TGF-β1 stimulation 8 h can be increased HKC cells ILK expression, and TGF-β1-induced increase in the synthesis of FN consistent. HKC cells transfected with pCMV-wtILK plasmid showed an increasing tendency of FN expression. Transfection of pCMV-kdILK inhibits the activity of ILK can significantly antagonize TGF-β1 stimulation HKC FN role. Conclusion The FN synthesis induced by TGF-β1 in renal tubular epithelial cells is closely related to the ILK expression induced by TGF-β1. Inhibition of ILK viability antagonizes FN synthesis by TGF-Bl.