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目的探讨高糖介导肾小管上皮细胞转分化及Ⅰ型胶原(Collagen Ⅰ)合成的分子机制。方法含35mmol/L葡萄糖培养液培养大鼠肾小管上皮细胞(NRK52E细胞),免疫化学方法检测p-Smad2/3核转位,ELISA检测细胞培养上清中TGF-β_1浓度,RT-PCR和Western blot方法分别检测α-SMA、E-cadherin、CollagenⅠ mRNA和蛋白的表达。观察TGF-β_1中和抗体对高糖上述效应的影响。结果高糖促进NRK52E细胞合成和分泌TGF-β_1。TGF-β_1中和抗体能抑制高糖介导的p-Smad2/3核转位,下调高糖介导的α-SMA和CollagenⅠ蛋白表达,上调E-Cadherin蛋白表达(P均<0.01)。结论高糖介导的肾小管上皮细胞转分化和细胞外基质Collagen Ⅰ的合成依赖于TGF-β_1效应。
Objective To investigate the molecular mechanism of high glucose - induced transdifferentiation of renal tubular epithelial cells and the synthesis of type Ⅰ collagen (Collagen Ⅰ). Methods Rat renal tubular epithelial cells (NRK52E cells) were cultured in 35 mmol / L glucose medium. The nuclear translocation of p-Smad2 / 3 was detected by immunochemistry. The concentration of TGF-β1 in the culture supernatants was detected by ELISA. blot were used to detect the mRNA and protein expression of α-SMA, E-cadherin and CollagenⅠ. To observe the effect of TGF-β 1 neutralizing antibody on the above effects of high glucose. Results High glucose promoted NRK52E cells to synthesize and secrete TGF-β_1. The neutralizing antibody of TGF-β_1 could inhibit the hyperglycemia-mediated nuclear translocation of p-Smad2 / 3, down-regulate the expression of α-SMA and CollagenⅠ, and up-regulate the expression of E-Cadherin (all P <0.01). Conclusion High glucose-induced renal tubular epithelial cell transdifferentiation and extracellular matrix Collagen Ⅰ synthesis depend on TGF-β 1 effect.