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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 investigate the role of early growth factor 1 (Egr-1) in renal tubular epithelial cell transdifferentiation. METHODS: (1) In vitro experiments: The expression of Egr-1 and transdifferentiation marker E (hTERT) was detected by qRT-PCR and Western blotting using pcDNA3.1, pcDNA3.1 / Egr- cadherin and fibroblast-specific protein (Fsp-1) mRNA and protein levels; cotransfection of HK-2 with pcDNA3.1 / Egr-1 and siEgr-1 or psilencer3.1, Western blot analysis Egr-1, E-cadherin, Fsp-1 protein expression changes. (2) In vivo experiments: The rat model of chronic renal fibrosis - subhepatectomy of 5/6 kidneys was established. Immunohistochemistry was used to detect the expression of Egr-1, E-cadherin and Fsp-1 expression. Results: (1) The expression of E-cad-herin in epithelial cells decreased and the expression of Fsp-1, a marker of fibroblasts, increased after HK-2 cells overexpression of Egr-1 <0.05), suggesting that the tubular epithelial cell transdifferentiation phenotype. However, after interfering Egr-1 with small interfering RNA, the transdifferentiation phenotype was reversed compared with empty vector group and parental cell group. (2) In vivo experiments showed that compared with the sham-operation group (SOR), the expression of E-cadherin and Fsp-1 in subepithelial nephrectomy of 5/6 kidneys were decreased and the expression of Fsp- -1 expression was significantly increased, suggesting that renal tubular epithelial cells transdifferentiation, and its expression of Egr-1 is closely related. Conclusion: Overexpression of Egr-1 promotes tubular epithelial cell transdifferentiation and is involved in the development of renal interstitial fibrosis.