| 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24 h和48 h,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。 |
| 关键词: TIEG RNA干扰 肾小管上皮细胞 Smad2 慢病毒载体 |
| DOI：10.3724/SP.J.1008.2010.0354 |
| 投稿时间：2009-03-30修订日期：2009-12-24 |
| 基金项目:国家自然科学基金(30470812),珠海市科技计划项目(PC20071006). |
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| Effect of siRNA targeting TGF-β inducible early gene on expression of advanced glycation end products-mediated Smad2 in renal tubular epithelial cells |
| YE Li-hong1, SHU Xiao-chun1, LU Hong-yun1, HU Fang1, SUN Liao1*, XIAO Hai-peng2* |
| (1. Department of Endocrinology, The Fifth Affiliated Hospital of Sun Yat-Sen University,Zhuhai 519000, Guangdong, China;2. Department of Endocrinology, The First Affiliated Hospital of Sun Yat-Sen University,Guangzhou 510600, Guangdong, China) |
| Abstract: |
| 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. |
| Key words: TIEG RNA interference tubular epithelial cells Smad2 lentiviral vector |
