Department of Pediatric Nephrology and Rheumatology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, PR China.
Mol Med Rep. 2013 May;7(5):1549-54. doi: 10.3892/mmr.2013.1365. Epub 2013 Mar 12.
Epithelial‑mesenchymal transition (EMT) is considered to be a crucial stage of renal fibrosis. Previous studies have indicated that paired box 2 (PAX2) affects renal fibrosis, likely by regulating EMT. Therefore, we examined whether PAX2 directly induces normal renal tubular EMT in vitro. Recombinant PAX2 plasmid was transfected into a renal tubular epithelial cell line (NRK52E) derived from normal rats. The optimal time‑point was evaluated through fluorescence detection, western blotting and real-time polymerase chain reaction (PCR). Morphological alterations were examined using phase‑contrast microscopy. At the optimal time‑point of transfection, the expression of E-cadherin, α-smooth muscle actin (α-SMA), fibronectin and snail was analyzed by western blotting and real-time PCR. PAX2 expression in NRK52E cells transfected with PAX2 plasmid reached its peak at 72 h post‑transfection. The transfected cells became elongated. PAX2 transfection induced a fibrotic phenotype of NRK52E cells with increased expression of α-SMA, fibronectin and snail as well as decreased expression of E-cadherin. The optimal time-point for transfection efficiency was 72 h. In conclusion, PAX2 induces renal tubular mesenchymal transition in vitro.
上皮-间充质转化（EMT）被认为是肾脏纤维化的关键阶段。先前的研究表明，配对盒基因 2（PAX2）影响肾脏纤维化，可能通过调节 EMT 来实现。因此，我们研究了 PAX2 是否可以直接在体外诱导正常肾小管 EMT。将重组 PAX2 质粒转染到正常大鼠来源的肾小管上皮细胞系（NRK52E）中。通过荧光检测、western blot 和实时聚合酶链反应（PCR）来评估最佳转染时间点。通过相差显微镜观察形态学变化。在转染的最佳时间点，通过 western blot 和实时 PCR 分析 E-钙黏蛋白、α-平滑肌肌动蛋白（α-SMA）、纤连蛋白和蜗牛的表达。转染 PAX2 质粒的 NRK52E 细胞中 PAX2 的表达在转染后 72 小时达到峰值。转染后的细胞变长。PAX2 转染诱导 NRK52E 细胞产生纤维化表型，α-SMA、纤连蛋白和蜗牛的表达增加，E-钙黏蛋白的表达减少。转染效率的最佳时间点为 72 小时。总之，PAX2 在体外诱导肾小管间充质转化。
