TIEGsiRNA对AGEs介导肾小管上皮细胞PAI-1表达的影响  

Effects of TIEG siRNA on the expression of PAI-1 induced by AGEs in renal tubular epithelial cells

作　　者：孟晓军[1] 舒晓春[1] 曾映娟[1] 文江华[1] 胡芳[1] 杨琼[1] 张瑜[1] 叶礼红[1] 孙辽[1] 

机构地区：[1]中山大学附属第五医院内分泌科,广东珠海519000

出　　处：《中国预防医学杂志》2011年第9期733-736,共4页Chinese Preventive Medicine

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

摘　　要：目的观察TIEG基因沉默对AGEs介导肾小管上皮细胞PAI-1表达的影响。方法以pshRNA-copGFP-lentivector作为载体,构建含有TIEG基因的慢病毒载体psiRNA-TIEG,转染至正常大鼠近端肾小管上皮细胞(NRK52E),然后给予AGEs刺激24 h和48 h,采用Western blot方法测定PAI-1蛋白的表达水平。结果 AGEs以时间依赖方式上调NRK52E细胞TIEG mRNA和PAI-1蛋白表达。TGF-β1中和抗体可有效降低AGEs刺激NRK52E细胞48 h后PAI-1的表达水平(1.150±0.089vs0.707±0.015,P<0.05)。TIEGsiRNA转染后可显著下调AGEs刺激NRK52E细胞48 h后TIEG mRNA表达水平(0.852±0.019vs0.448±0.028,P<0.01),及PAI-1蛋白表达水平(0.396±0.042vs0.245±0.063,P<0.05)。结论 TIEG基因沉默能有效抑制AGEs诱导NRK52E细胞PAI-1的表达。Objective To investigate the effects of TIEG gene silencing on the expression of PAI-1 induced by AGEs in renal tubular epithelial cells.Methods The plasmid of psiRNA-TIEG was a TIEG gene containing pshRNA-copGFP-lentivector.The reconstructed vector was transfected into normal rat proximal tubular epithelial cells（NRK52E） and cultured in medium with AGEs-BSA for 24hrs or 48hrs.The expression of PAI-1 was measured by Western blotting.Results The concentrations of TIEG mRNA and PAI-1 were time-dependently upregulated by AGEs in NRK52E cells.The enhanced expression of PAI-1 by AGEs stimulation was effectively inhibited by TGF-β1 antibody at 48 hrs in NRK52E cells（1.150±0.089 vs 0.707±0.015,P0.05）.TIEG siRNA effectively down-regulated the AGEs-stimulated production of TIEG mRNA（0.448±0.028 vs 0.852±0.019,P0.01）and PAI-1（0.245±0.063 vs 0.396±0.042,P0.05）at 48 hrs in NRK52E cells.Conclusion The expression of PAI-1 induced by AGEs could be effectively inhibited by TIEG gene silencing in NRE52E cells.

关 键 词：TIEG 基因沉默 肾小管上皮细胞 血浆纤溶酶原激活物抑制剂-1(PAI-1) 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]