早期生长反应因子1过表达促进肾小管上皮细胞转分化  

Overexpression of Egr-1 promotes epithelial-to-mesenchymal transition on renal tubular epithelial cells

作　　者：翟英[1] 孙世仁[1] 杜锐[1] 黄晨[1] 王汉民[1] 

机构地区：[1]第四军医大学西京医院肾脏内科,西安710032

出　　处：《肾脏病与透析肾移植杂志》2009年第6期537-541,共5页Chinese Journal of Nephrology,Dialysis & Transplantation

摘　　要：目的:探讨早期生长反应因子1(Egr-1)在肾小管上皮细胞转分化中的作用。方法:(1)体外实验:用pcDNA3.1,pcDNA3.1/Egr-1转染肾小管上皮细胞株HK-2,qRT-PCR法、Western免疫印迹法检测Egr-1及转分化标志物E-cadherin、成纤维细胞特异性蛋白(Fsp-1)mRNA及蛋白水平的表达变化;用pcDNA3.1/Egr-1和siEgr-1或psilencer3.1共转染HK-2,Western免疫印迹法检测Egr-1、E-cadherin、Fsp-1蛋白水平的表达变化。(2)体内实验:建立大鼠慢性肾纤维化模型——5/6肾脏次全切除,造模成功后免疫组化法检测大鼠肾组织中Egr-1、E-cadherin、Fsp-1的表达。结果:(1)与转染空载体相比,HK-2细胞过表达Egr-1后,上皮细胞标志性蛋白E-cad-herin表达减弱,成纤维细胞标志性蛋白Fsp-1表达增强(P<0.05),提示出现肾小管上皮细胞转分化表型。而用小干扰RNA干扰掉Egr-1后,与空载体组及亲本细胞组相比,转分化表型得到逆转。(2)体内实验证实,与假手术组(SOR)相比,5/6肾脏次全切除肾组织中,转分化标志蛋白E-cadherin表达减弱、Fsp-1表达增加,同时肾小管上皮细胞Egr-1的表达明显升高,提示肾小管上皮细胞发生转分化,且其与Egr-1过表达密切相关。结论:Egr-1的过表达促进了肾小管上皮细胞转分化,参与肾间质纤维化的发生发展。Objective：To explore the role of early growth response factor-1 （Egr-1） on epithelial-to-mesenchymal transition（EMT） of renal tubular epithelial cells. Methodology： （ 1 ） Transfected the plasmids pcDNA3. 1/Egr-1 and empty vector pcDNA3.1 into ttK-2 cells with Lipofeetamine 2000 in vitro. Real-time PCR and Western Blot analyses were used to detect the expression of Egr-1, E-eadherin, Fsp-1 in mRNA and protein levels. Western blot analysis was used to detect Egr-1, E-cadberin, and Fsp-1 expression in parental cells, pSilencer3.1 empty vector-transfected cells, and Egr-1 siRNA transfected cells. （2） After successfully establishing the animal model of the 5/6 subtotal nephrectomy, the expression of Egr-1, E-cadherin, and Fsp-1 in renal tissues was also determined though immunohistochemistry analysis. Results ： （ 1 ） Overexpression of Egr-1 significantly reduced the expression of the epithelial marker E-cadherin and enhanced the expression of the fibroblastic marker Fsp-1 （ P 〈 0. 05 ） , while cotransfection with Egr-1 siRNA reversed the phenotype of EMT. （2） Immanohistocheistry examination showed that the staining of E-cadherin was decreased, whereas Fsp-1 was increased on renal tubular epithelial cells in 5/6 subtotal nephrectomy compared with that in SOR. Meanwhile, Egr-1 was also highly expressed in 5/6-nephrectmized kidney tissue, whereas almost no staining in SOR kidney. Conclusion： Overexpression of Egr-1 promotes renal EMT through modulating the expression of E-cadherin and Fsp-1.

关 键 词：上皮细胞向间质转分化 早期生K反应因子 E钙粘素 成纤维细胞特异性蛋白 

分 类 号：R692[医药卫生—泌尿科学]