慢病毒介导的SHP-1过表达抑制模型小鼠动脉粥样硬化  

Overexpression of SHP-1 mediated by lentivirus restrains atherosclerotic progression in mice

作　　者：张兵兵[1] 宋恒良[1] 孙涛[2] 万大国[1] 

机构地区：[1]郑州大学第二附属医院心血管内科,河南郑州450014 [2]河南省人民医院心血管内科,河南郑州450003

出　　处：《中国病理生理杂志》2015年第8期1395-1400,共6页Chinese Journal of Pathophysiology

摘　　要：目的:研究含SH2结构域的酪氨酸磷酸酶-1(SHP-1)慢病毒载体在动脉粥样硬化模型小鼠中的作用。方法:将45只8周龄雄性Apo E基因敲除小鼠随机分为3组,即对照组、绿色荧光蛋白(GFP)转染组、SHP-1转染组。3组小鼠右侧颈总动脉植入套环并高脂喂养8周,随后分别转染GFP空载体和SHP-1慢病毒(SHP-1-LV)。分别于转染第1、2、6周检测硬化斑块荧光强度、小鼠体重、血清总胆固醇(TC)、甘油三酯(TG)水平变化,并利用real-time PCR和Western blot检测右侧颈总动脉中SHP-1、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶(MMP)-2、MMP-9等的表达,最后制作切片染色观察斑块病理变化。结果:荧光显微镜下观察到慢病毒转染第1、2、6周后斑块有明显荧光,且在第2周时荧光强度最高,SHP-1慢病毒转染对小鼠体重和血清TC、TG水平无显著影响,但可显著上调右侧颈总动脉中SHP-1的mRNA和蛋白的表达,同时抑制IL-6、TNF-α、MMP-2、MMP-9等的表达水平。SHP-1慢病毒转染也降低右侧颈总动脉斑块面积比及脂质含量。结论:过表达SHP-1可能促进小鼠动脉粥样硬化斑块消退,从而为预防和治疗动脉粥样硬化提供靶点。AIM： To investigate the role of SH2-domain-containing protein-tyrosine phosphatase-1（ SHP-1）lentivirus in atherosclerotic mice. METHODS： Apo E knock-out mice were randomly assigned to 3 groups： control group,GFP transfection group and SHP-1 transfection group. All mice were placed with carotid collars on the right common carotid arteries near its bifurcation,following feeding with high-fat diet for 8 weeks and then transfected with GFP blank vector or SHP-1 lentivirus（ SHP-1-LV）. The fluorescence density of the plaques,body weight,the levels of plasma total cholesterol（ TC） and triglyceride（ TG） were determined at 1st,2nd,and 6th week after lentivirus transfection. Furthermore,the mRNA and protein expression of SHP-1,interleukin-6（ IL-6）,tumor necrosis factor-α（ TNF-α）,matrix metalloproteinase（ MMP）-2 and MMP-9 were analyzed by real-time PCR and Western blot. Additionally,pathological analysis of the plaques was also performed by HE and oil red O staining. RESULTS： The fluorescence of the plaques was observed at 1st,2nd,and 6th week after lentivirus transfection,with a highest density at 2nd week. The body weight and the levels of TC and TG in the mice were not influenced by lentivirus transfection. Moreover,SHP-1-LV transfection significantly upregulated the expression of SHP-1 at mRNA and protein levels,but inhibited the expression of IL-6,TNF-α,MMP-2 and MMP-9. In addition,SHP-1-LV transfection also decreased the plaque size ratio and lipid content in right common carotid arteries.CONCLUSION： SHP-1 overexpression accelerates the regression of atherosclerotic plaque,thus emerging SHP-1 as a target for prevention and treatment of atherosclerosis.

关 键 词：含SH2结构域的酪氨酸磷酸酶-1 动脉粥样硬化 慢病毒 APOE基因敲除小鼠 

分 类 号：R543[医药卫生—心血管疾病]