中文摘要：

目的研究细胞外信号调节激酶2（ERK2）信号转导通路对结缔组织生长因子（CTGF）诱导人肾小管上皮细胞（HK-2）向间充质细胞转化的调控效应。方法腺病毒介导的反义ERK2基因（Adanti-ERK2）按感染强度50、100、200转染HK-2细胞,确定最适感染强度。按作用因素的不同将HK-2细胞分为对照组、Ad-LacZ组、CTGF组和Adanti-ERK2组。免疫组织化学检测E-Cadherin和Vi mentin的表达,Western blot法检测E-Cadherin、Vi mentin及ERK2的表达变化;细胞接种Boyden小室1、3、5 d时检测各组HK-2细胞迁移数目的变化。结果①Adanti-ERK2转染HK-2细胞最适感染强度为100。②HK-2细胞在CTGF刺激下E-Cadherin蛋白表达明显减少,Vi mentin表达增加,同时细胞内的ERK2蛋白表达显著增加;而Adanti-ERK2组E-Cadherin和Vi mentin表达变化不明显,ERK2蛋白表达较对照组明显减少。③接种5 d,HK-2细胞在CTGF刺激下细胞运动能力显著高于其他各组,而Adanti-ERK2组与对照组和Ad-LacZ组相比差异无显著性意义。结论以反义ERK2基因治疗可以有效阻断CTGF诱导的肾小管上皮细胞向间充质转化。提示ERK2信号转导通路在该转化中发挥重要作用。

英文摘要：

Objective To investigate the effects of ERK2 signal transduction pathway on the epithelial mesenchymal transition of HK-2 cells induced by connective tissue growth factor（CTGF） in vitro. Methods ①The HK-2 cells were transfeeted by adenovirus-mediated antisense ERK2（Adanti-ERK2） with multiplicity of infection（MOI） of 50,100 and 200, and observed by green fluorescence 2 days later. The ratio of live cells was measured at the 2nd day after transfection. ②The cultured HK-2 cells were divided into 4 groups：control group,Ad-LacZ group, CTGF group and Adanti-ERK2 group. Immunohistochemistry was used to detect the expression of E-Cadherin and Vimentin in the cells. Western-blot was used to detect the E-Cadherin,Vimentin and ERK2 proteins. Boyden Chamber was used to test the migration of HK-2 cells at first, third, and 5th day after transfection. Results ① The optimal MOI of HK-2 cells transfected by Adanti-ERK2 was 100. ② The HK-2 cells expressed more Vimentin,ERK2 and less E-Cadherin with the stimulation of CTGF. There was no significant difference in the expression of E-Cadherin and Vimentin among control group, Ad-LacZ group and Adanti-ERK2 group, but the expression of ERK2 protein in Adanti-ERK2 group was down-regulated markedly. ③ At the 5th day after transfection, the HK-2 cells were transformed and obtained motility induced by CTGF. However,there was no significant difference in the migration of the cells among gene ther- apy group,empty control group and vector control group. Conclusion In vitro ,Adanti-ERK2 gene therapy can inhibit the tubular epithelial mesenchymal transition induced by CTGF, suggesting ERK2 signal transduction pathway may play an important role.