TIEG特异siRNA对糖基化终末产物介导的肾小管上皮细胞Smad2表达的影响  

Effect of siRNA targeting TGF-β inducible early gene on expression of advanced glycation end products-mediated Smad2 in renal tubular epithelial cells

作　　者：叶礼红[1] 舒晓春[1] 鲁红云[1] 胡芳[1] 孙辽[1] 肖海鹏[2] 

机构地区：[1]中山大学附属第五医院内分泌科,珠海519000 [2]中山大学附属第一医院内分泌科,广州510600

出　　处：《第二军医大学学报》2010年第4期354-358,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30470812);珠海市科技计划项目(PC20071006)~~

摘　　要：目的以RNA干扰技术沉默TGF-β诱导早期基因(TIEG)的表达,观察TIEG沉默对糖基化终末产物(AGEs)介导的肾小管上皮细胞Smad2表达的影响。方法以pshRNA-copGFP-lentivector作为载体,构建含TIEG特异siRNA的慢病毒载体psiRNA-TIEG,将其转染至大鼠近端肾小管上皮细胞(NRK52E),然后给予糖基化修饰的牛血清白蛋白(AGE-BSA)刺激24h和48h,采用PCR方法测定其TIEG mRNA、Smad2 mRNA表达水平,蛋白免疫印迹法测定Smad2蛋白表达水平。以转染空质粒载体组作为对照。结果TIEG特异的siRNA可有效下调AGEs诱导的TIEG mRNA、Smad2 mRNA和蛋白表达,与空质粒载体组相比差异有统计学意义(P<0.05)。结论TIEG的沉默能有效抑制AGEs介导的NRK52E细胞Smad2 mRNA和蛋白的表达。Objective To investigate the effect of siRNA targeting TGF-β inducible early gene（TIEG） on expression of Smad2 in advanced glycation end products（AGEs）-mediated renal tubular epithelial cells.Methods The pshRNA-copGFP-lentivector containing target gene was used as a vector to construct siRNA-TIEG,which was then transfected into normal rat proximal tubular epithelial cells（NRK52E）,which were then cultured in RPMI 1640 medium supplemented with AGE-BSA for 24 h and 48 h.The expression of Smad2 mRNA and protein was examined by PCR,Western blotting analysis,respectively.Blank vector group served as control.Results SiRNA-TIEG significantly reduced the expression of TIEG mRNA,Smad2 mRNA and Smad2 protein in normal rat proximal tubular epithelial cells in presence of AGE-BSA compared with those in the control group（P 0.05）.Conclusion Silence of TIEG can effectively inhibit the expression of Smad2 mediated by AGEs in NRK52E cells.

关 键 词：TIEG RNA干扰 肾小管上皮细胞 SMAD2 慢病毒载体 

分 类 号：R587.24[医药卫生—内分泌]