中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
8期
1395-1400
,共6页
张兵兵%宋恒良%孙涛%万大国
張兵兵%宋恆良%孫濤%萬大國
장병병%송항량%손도%만대국
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
含SH2結構域的酪氨痠燐痠酶-1%動脈粥樣硬化%慢病毒%ApoE基因敲除小鼠
함SH2결구역적락안산린산매-1%동맥죽양경화%만병독%ApoE기인고제소서
SH2-domain-containingprotein-tyrosinephosphatase-1%Atherosclerosis%Lentivirus%ApoEknock-out mice
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。
目的:研究含SH2結構域的酪氨痠燐痠酶-1(SHP-1)慢病毒載體在動脈粥樣硬化模型小鼠中的作用。方法:將45隻8週齡雄性ApoE基因敲除小鼠隨機分為3組,即對照組、綠色熒光蛋白( GFP)轉染組、SHP-1轉染組。3組小鼠右側頸總動脈植入套環併高脂餵養8週,隨後分彆轉染GFP空載體和SHP-1慢病毒( SHP-1-LV)。分彆于轉染第1、2、6週檢測硬化斑塊熒光彊度、小鼠體重、血清總膽固醇( TC)、甘油三酯( TG)水平變化,併利用real-time PCR和Western blot檢測右側頸總動脈中SHP-1、白細胞介素-6(IL-6)、腫瘤壞死因子-α(TNF-α)、基質金屬蛋白酶( MMP)-2、MMP-9等的錶達,最後製作切片染色觀察斑塊病理變化。結果:熒光顯微鏡下觀察到慢病毒轉染第1、2、6週後斑塊有明顯熒光,且在第2週時熒光彊度最高,SHP-1慢病毒轉染對小鼠體重和血清TC、TG水平無顯著影響,但可顯著上調右側頸總動脈中SHP-1的mRNA和蛋白的錶達,同時抑製IL-6、TNF-α、MMP-2、MMP-9等的錶達水平。 SHP-1慢病毒轉染也降低右側頸總動脈斑塊麵積比及脂質含量。結論:過錶達SHP-1可能促進小鼠動脈粥樣硬化斑塊消退,從而為預防和治療動脈粥樣硬化提供靶點。
목적:연구함SH2결구역적락안산린산매-1(SHP-1)만병독재체재동맥죽양경화모형소서중적작용。방법:장45지8주령웅성ApoE기인고제소서수궤분위3조,즉대조조、록색형광단백( GFP)전염조、SHP-1전염조。3조소서우측경총동맥식입투배병고지위양8주,수후분별전염GFP공재체화SHP-1만병독( SHP-1-LV)。분별우전염제1、2、6주검측경화반괴형광강도、소서체중、혈청총담고순( TC)、감유삼지( TG)수평변화,병이용real-time PCR화Western blot검측우측경총동맥중SHP-1、백세포개소-6(IL-6)、종류배사인자-α(TNF-α)、기질금속단백매( MMP)-2、MMP-9등적표체,최후제작절편염색관찰반괴병리변화。결과:형광현미경하관찰도만병독전염제1、2、6주후반괴유명현형광,차재제2주시형광강도최고,SHP-1만병독전염대소서체중화혈청TC、TG수평무현저영향,단가현저상조우측경총동맥중SHP-1적mRNA화단백적표체,동시억제IL-6、TNF-α、MMP-2、MMP-9등적표체수평。 SHP-1만병독전염야강저우측경총동맥반괴면적비급지질함량。결론:과표체SHP-1가능촉진소서동맥죽양경화반괴소퇴,종이위예방화치료동맥죽양경화제공파점。
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 .