慢病毒介导的 SH P－1过表达抑制模型小鼠动脉粥样硬化

摘要

AIM:ToinvestigatetheroleofSH2-domain-containingprotein-tyrosinephosphatase-1(SHP-1) lentivirus in atherosclerotic mice .METHODS:ApoE 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 tar-get for prevention and treatment of atherosclerosis .%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ApoE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。