沉默转化生长因子β诱导基因对糖尿病大鼠肾组织内Smad2、P-smad2及Ⅳ型胶原表达的影响  

Effect of TIEG-siRNA on Smad2, p-smad2 and collagen Ⅳ in diabetic nephropathy

作　　者：舒晓春[1] 尹代婵[1] 叶礼红[1] 胡芳[1] 文江华[1] 杨琼[1] 孙辽[1] 

机构地区：[1]中山大学附属第五医院内分泌科,广东省珠海市519000

出　　处：《中华医学杂志》2010年第26期1859-1862,共4页National Medical Journal of China

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 探讨沉默转化生长因子β诱导基因（TIEG-siRNA）对糖尿病大鼠肾组织Smad2、p-shred2及Ⅳ型胶原表达的影响.方法 链脲佐菌素诱导的糖尿病大鼠模型10只,分为空载体组和TIEG-siRNA治疗组,另以5只正常大鼠作为对照组.治疗组和空载体组分别给予TIEG-siRNA病毒液和空载体质粒0.5 ml,于0、72 h两次尾静脉注射,于成模后4周处死.实时荧光定量PCR检测大鼠肾组织内TIEG-mRNA的表达水平,免疫组织化学检测肾组织Smad2、p-smad2、Ⅳ型胶原蛋白表达.Western印迹检测大鼠肾组织Smad2,p-smad2蛋白表达水平.结果 TIEG-mRNA表达在TIEG-siRNA 治疗组（0.0636±0.0066）较空载体组（0.1054±0.0111）明显下调（P〈0.05）,免疫组化半定量结果示Smad2蛋白表达在治疗组[（2.13±0.19）%]较空载体组[（2.53±0.34）%]明显减少（P〈0.05）,p-smad2蛋白和Ⅳ型胶原表达在TIEG-siRNA治疗组较空载体组明显减少[（21.77±2.00）%比（27.03±2.51）%,（3.67士0.42）%比（4.85±0.43）%,P〈0.05].Western印迹显示Smad2在治疗组比空载体组明显减少（0.32±0.09比0.50±0.04,P〈0.05）.P-smad2在治疗组比空载体组明显减少（0.16±0.01比0.32±0.02,P〈0.05）.结论 TIEG-siRNA可以通过影响糖尿病大鼠肾脏内Smad2表达及其活化,从而下调Ⅳ型胶原表达来延缓糖尿病纤维化进程.Objective To investigate the effect of TIEG-siRNA on Smad2, p-smad2 and collagen Ⅳ in diabetic nephropathy rats induced by streptozotocin （STZ） . Methods Ten Sprague-Dawley rats injected with STZ were randomly divided into TIEG-siRNA and Control groups. Other five normal rats were used as control. Each of the TIEG-siRNA and Control groups were injected with TIEG-siRNA and Control 0. 5 ml via tail vein at 0 and 72 hours respectively. All rats were sacrificed at week 4 after a successful modeling. To confirmed the efficacy of TIEG-siRNA in rat kidney, the TIEG levels were determined by fluorescence quantitative PCR. The expressions of Smad2, p-smad2 and collagen Ⅳ protein were detected by immunohistochemical method while Smad2 and p-smad2 examined by Western blot Results The TIEG levels were greatly down-regulated in the TIEG-siRNA treated group （0.0636 ± 0.0066） versus the empty vector treated group （0. 1054 ± 0. 0111） （ P 〈 0. 05）. The immunohistochemical semi-quantitative method indicated that there was a decrease in the TIEG-siRNA treated group versus the empty vector treated group：（2. 13 ±0.19）% vs （2.53 ±0.34）% in Smad2, （21. 77 ± 2. 00）% vs （27. 03 ±2. 51）% in p-smad2 and （3.67 ±0.42）% vs （4. 85 ±0.43） % in collagen Ⅳ. Western blot also showed that smad2 in the TIEG-siRNA treated group （0. 32 ± 0. 09 ） was much lower than that in the empty vector treated group （0. 50 ±0. 04）. And p-smad2 in the TEEG-siRNA treated group（0. 16 ±0. 01）was much lower than that in the empty vector treated group （ 0. 32 ± 0. 02 ） （P 〈 0. 05）. Conclusion TIEG-siRNA may be useful in preventing the progression of diabetic nephropathy through influencing the expression of Smad2 and its activation and down-regulating collagen Ⅳ.

关 键 词：糖尿病肾病 转化生长因子Β 基因 

分 类 号：R587.1[医药卫生—内分泌]