细胞因子信号转导抑制剂3在大鼠静脉移植物中的表达及意义  

Expression and significance of suppressor of cytokine signaling 3 in vein grafts of rat model

作　　者：黄进启 姚元波 向水 蔡彦力 郑勇 刘金平[2] 

机构地区：[1]湖北省恩施土家族苗族自治州中心医院心胸外科,湖北恩施445000 [2]华中科技大学同济医学院附属协和医院心血管外科,湖北武汉430022

出　　处：《中国医药导报》2015年第22期27-32,共6页China Medical Herald

基　　金：湖北省卫生和计划生育委员会科研项目(WJ2015MB192)

摘　　要：目的检测细胞因子信号转导抑制剂3(SOCS3)在大鼠颈外静脉颈总动脉移植物和体外培养的动脉血管平滑肌细胞增殖模型中的表达,探讨其在静脉移植物再狭窄病理过程中的可能作用及机制。方法体内实验分两组,将36只雄性SD大鼠随机分成实验组和对照组,每组18只。实验组采用标准显微外科技术行颈外静脉颈总动脉翻转端吻合,对照组行假手术,分别于术后第1、3、7天取出移植静脉。体外培养的大鼠血管平滑肌细胞用碱性成纤维细胞生长因子(b FGF)刺激,再用大鼠SOCS3基因重组腺病毒(p Yr Ad-r SOCS3)、对照病毒p Yr Ad-GFP转染,体外实验分四组:对照组,b FGF组,b FGF+p Yr Ad-GFP组,b FGF+p Yr Ad-r SOCS3组。应用Real time-PCR和Western boltting检测白介素6(IL-6)、单核细胞趋化蛋白1(MCP-1)、肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)、细胞间黏附分子1(ICAM-1)、信号转导与转录激活因子(STAT3)、磷酸化的信号转导与转录激活因子(PSTAT3)、SOCS3 m RNA和蛋白表达水平。结果体内实验表明,与对照组比较,实验组静脉移植物中IL-6[(3.60±0.51)比(1.00±0.00);(1.52±0.37)比(0.35±0.05)]、MCP-1[(2.08±0.38)比(1.00±0.00);(1.90±0.31)比(0.85±0.17)]、TNF-α[(4.86±0.74)比(1.00±0.00);(1.66±0.30)比(0.29±0.07)]、IL-1β[(2.73±0.52)比(1.00±0.00);(0.74±0.17)比(0.19±0.04)]、ICAM-1[(1.97±0.35)比(1.00±0.00);(1.02±0.39)比(0.21±0.02)]、SOCS3[(1.93±0.38)比(1.00±0.00);(0.82±0.18)比(0.42±0.12)]m RNA和蛋白表达水平以及STAT3[(1.50±0.36)比(0.21±0.05)]、P-STAT3[(1.54±0.39)比(0.37±0.10)]蛋白表达水平均在1周内明显升高(P<0.05或P<0.01)。体外实验结果表明,与对照组比较,b FGF组中上述指标m RNA和蛋白表达明显上调(P<0.05或P<0.01);与b FGF组比较,b FGF+p Yr Ad-r SOCS3组中SOCS3 m RNA和蛋白[(5.47±1.03)比(1.37±0.24);(1.79±0.38)比(1.28±0.32)]表达进一步上调(P<0.01、P<0.05),而IL-6[(1.28±0.25)比(1.57±0.31);(1.68±0.39)比(2.36±Objective To detect the expression of suppressor of cytokine signaling 3（SOCS3） in vein grafts of rat and explore its possible effect and mechanism in vein graft stenosis. Methods The experiment in vivo was divided into two groups. Thirty-six male Sprague-Dawley rats were randomly divided into two groups： experimental group and control group, with 18 cases in each group.Autologous jugular vein to carotid artery reverse interposition grafts were performed by standard microsurgical technique in rats of experimental group. Sham operation was performed in rats of control group. The vein grafts were harvested on day 1, day 3, and day 7 after operation. The rat vascular smooth muscle cells were stimulated with basic fibroblast growth factor（b FGF） and then transfected with the recombinant adenovirus carrying the rat SOCS3 gene（p Yr Ad-r SOCS3）, the matched control adenovirus vector（p Yr Ad-GFP）, which carries only GFP. Experiments included the following groups： control group, b FGF group, b FGF＋p Yr Ad-GFP group, b FGF＋p Yr Ad-r SOCS3 group. The harvested vein grafts and VSMCs were analyzed by real time-PCR and Western blot to detect the m RNA and protein expression levels of interleukin 6（IL-6）, monocyte chemotactic protein（MCP）-1, tumor necrosis factor（TNF）-α, interleukin-1β（IL-1β）, intercellular adhension molecular（ICAM）-1, signal transducer and activator of transcription（STAT3）, P-STAT3, SOCS3. Results In vivo, compared with control group, the m RNA and protein expression levels of IL-6 [（3.60±0.51） vs（1.00 ±0.00）;（1.52 ±0.37） vs（0.35 ±0.05）], MCP-1 [（2.08 ±0.38） vs（1.00 ±0.00）;（1.90 ±0.31） vs（0.85 ±0.17）],TNF-α [（4.86 ±0.74） vs（1.00 ±0.00）;（1.66 ±0.30） vs（0.29 ±0.07）], IL-1β [（2.73 ±0.52） vs（1.00 ±0.00）;（0.74 ±0.17） vs（0.19 ±0.04）], ICAM-1 [（1.97 ±0.35） vs（1.00 ±0.00）;（1.02 ±0.39） vs（0.21 ±0.02）], SOCS3 [（1.93 ±0.38） vs（1.00 ±0.00）;（0.82±0.18） vs（

关 键 词：细胞因子信号转导抑制剂3 信号转导与转录激活因子 炎症 静脉移植物 

分 类 号：R965[医药卫生—药理学]