CAJ | 학술논문

目的探讨以胎儿肉瘤样酪氨酸激酶1(FLT-1)为启动子携带目的基因蛋白C(PC)的重组腺病毒(Ad-FLT-1/PC)对糖尿病肾病动脉粥样硬化(AS)大鼠血管壁炎症因子表达的影响.方法将68只糖尿病肾病造模成功的AS大鼠随机分为Ad-FLT-1/PC组(23只)、带有绿色荧光蛋白(GFP)空腺病毒(Ad-GFP)组(23只)以及生理盐水(NS)组(22只),分别经尾静脉注射滴度约2.1 ×1010pfu/ml的Ad-FLT-1/PC 300μl、滴度约1.3×1010pfu/ml的空腺病毒300μl及无菌生理盐水300μl,以正常SD大鼠(23只)作为对照组.分别在转染1、3、7、14 d处死大鼠,快速冰冻切片观察血管壁绿色荧光分布,用免疫组织化学(IHC)方法检测PC蛋白在血管壁上的表达情况;IHC检测血管壁肿瘤坏死因子α(TNF-α)、细胞间黏附分子1(ICAM-1)蛋白表达水平的变化.结果尾静脉注射重组腺病毒Ad-FLT-1/PC后第3天可观察到绿色荧光在血管内皮层表达,且荧光至少能持续表达至第14天;转染3d后,Ad-FLT-1/PC组血管内皮层PC表达水平明显高于Ad-GFP组与NS组[(0.065±0.030)比(0.013 ±0.005)、(0.016 ±0.011),均P＜0.05];转染7d后,Ad-FLT-1/PC组血管壁的TNF-α、ICAM-1蛋白表达水平显著低于Ad-GFP组与NS组(均P＜0.05);Ad-GFP组与NS组各时间点蛋白表达水平差异均无统计学意义(均P＞0.05).结论糖尿病肾病AS大鼠经尾静脉注射Ad-FLT-I/PC后,PC可在血管内皮细胞有效表达并下调炎症因子的表达.
목적 탐토이태인육류양락안산격매1(FLT-1)위계동자휴대목적기인단백C(PC)적중조선병독(Ad-FLT-1/PC)대당뇨병신병동맥죽양경화(AS)대서혈관벽염증인자표체적영향.방법 장68지당뇨병신병조모성공적AS대서수궤분위Ad-FLT-1/PC조(23지)、대유록색형광단백(GFP)공선병독(Ad-GFP)조(23지)이급생리염수(NS)조(22지),분별경미정맥주사적도약2.1 ×1010pfu/ml적Ad-FLT-1/PC 300μl、적도약1.3×1010pfu/ml적공선병독300μl급무균생리염수300μl,이정상SD대서(23지)작위대조조.분별재전염1、3、7、14 d처사대서,쾌속빙동절편관찰혈관벽록색형광분포,용면역조직화학(IHC)방법검측PC단백재혈관벽상적표체정황;IHC검측혈관벽종류배사인자α(TNF-α)、세포간점부분자1(ICAM-1)단백표체수평적변화.결과 미정맥주사중조선병독Ad-FLT-1/PC후제3천가관찰도록색형광재혈관내피층표체,차형광지소능지속표체지제14천;전염3d후,Ad-FLT-1/PC조혈관내피층PC표체수평명현고우Ad-GFP조여NS조[(0.065±0.030)비(0.013 ±0.005)、(0.016 ±0.011),균P＜0.05];전염7d후,Ad-FLT-1/PC조혈관벽적TNF-α、ICAM-1단백표체수평현저저우Ad-GFP조여NS조(균P＜0.05);Ad-GFP조여NS조각시간점단백표체수평차이균무통계학의의(균P＞0.05).결론 당뇨병신병AS대서경미정맥주사Ad-FLT-I/PC후,PC가재혈관내피세포유효표체병하조염증인자적표체.
Objective To explore the effect of recombinant adenovirus Ad-FLT-1/PC on the expression of vascular inflammatory factors in rats with diabetic nephropathy atherosclerosis.Methods A total of 68 rats of diabetic nephropathy atherosclerosis were randomly dividcd into three groups of Ad-FLT-1/ PC [recombinant adenovirus with protein C (PC) gene,n =23],Ad-green fluorescence protein (recombinant blank adenovirus with GFP,n =23),normal saline (n =22) and compared with normal control (n =23).And 300 μl of each was transfected via caudal vein.At day l,3,7 and 14 posttransfection,5 or 6 rats of each group were randomly sacrificed to observe the distribution of vascular fluorescence by frozen section.And the expression of PC was examined by immunohistochemistry (IHC).The expressions of tumor necrosis factor-alpha (TNF-α) and intercellular adhesion molecule 1 (ICAM-1) protein in vascular wall were measured by IHC.Results At day 3 after an injection of Ad-FLT-1/PC,green fluorescence was observed in vascular endothelium and continued until 14 days.IHC showed protein C in endothelial cells of Ad-FLT-1/PC group was higher than those of Ad-GFP and NS groups at Days 3-14 (P ＜0.05).IHC showed the amounts of TNF-α and ICAM-1 in Ad-FLT-1/PC group were lower than those in GFP and NS groups at Day 7 post-transfection (P ＜ 0.05).GFP and NS groups had no difference at all timepoints (P ＞ 0.05).Conclusions The recombinant adenovirus Ad-FLT-1/PC may be transfected into vascular walls of rats with diabetic nephropathy atherosclerosis.And exogenous PC can effcctively express and protect vascular walls by inhibiting the expression of inflammatory cytokines.