CAJ | 학술논문

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.
목적 탐토침묵전화생장인자β유도기인(TIEG-siRNA)대당뇨병대서신조직Smad2、p-shred2급Ⅳ형효원표체적영향.방법 련뇨좌균소유도적당뇨병대서모형10지,분위공재체조화TIEG-siRNA치료조,령이5지정상대서작위대조조.치료조화공재체조분별급여TIEG-siRNA병독액화공재체질립0.5 ml,우0、72 h량차미정맥주사,우성모후4주처사.실시형광정량PCR검측대서신조직내TIEG-mRNA적표체수평,면역조직화학검측신조직Smad2、p-smad2、Ⅳ형효원단백표체.Western인적검측대서신조직Smad2,p-smad2단백표체수평.결과 TIEG-mRNA표체재TIEG-siRNA 치료조(0.0636±0.0066)교공재체조(0.1054±0.0111)명현하조(P<0.05),면역조화반정량결과시Smad2단백표체재치료조[(2.13±0.19)%]교공재체조[(2.53±0.34)%]명현감소(P<0.05),p-smad2단백화Ⅳ형효원표체재TIEG-siRNA치료조교공재체조명현감소[(21.77±2.00)%비(27.03±2.51)%,(3.67사0.42)%비(4.85±0.43)%,P<0.05].Western인적현시Smad2재치료조비공재체조명현감소(0.32±0.09비0.50±0.04,P<0.05).P-smad2재치료조비공재체조명현감소(0.16±0.01비0.32±0.02,P<0.05).결론 TIEG-siRNA가이통과영향당뇨병대서신장내Smad2표체급기활화,종이하조Ⅳ형효원표체래연완당뇨병섬유화진정.
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.