Ad-FLT-1/PC对糖尿病肾病动脉粥样硬化大鼠炎症因子表达的影响  

Effects of Ad-FLT-1/PC on the expression of inflammatory factors in rats with diabetic nephropathy atherosclerosis

作　　者：陈娅[1] 丁涵露[2] 王莉[2] 李贵森[2] 张萍[2] 汪伟[2] 

机构地区：[1]遵义医学院附属医院神经内科,贵州563003 [2]四川省医学科学院四川省人民医院肾内科

出　　处：《中华医学杂志》2015年第24期1961-1965,共5页National Medical Journal of China

基　　金：四川省科技厅支撑计划（2010JY0110）

摘　　要：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 ×10^10pfu/ml的Ad-FLT-1/PC 300μl、滴度约1.3×10^10pfu/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.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.

关 键 词：糖尿病肾病 动脉粥样硬化 蛋白C 重组腺病毒 

分 类 号：R587.2[医药卫生—内分泌]