近红外荧光分子标记的LOX-1抗体靶向小鼠动脉粥样硬化模型的成像研究
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摘要
目的:探讨近红外荧光分子标记的凝集素样氧化低密度脂蛋白受体-1(LOX-1)抗体探针在ApoE-/-鼠主脉动脉粥样硬化近红外荧光(NIRF)成像中的可行性。方法:将ApoE-/-小鼠以高脂饮食喂养20周,建立动脉粥样硬化模型。将24只ApoE-/-动脉粥样硬化模型小鼠随机分为3组,即抗LOX-1抗体-NIR-797组、非特异性IgG-NIR-797组和单纯磷酸盐缓冲液(PBS)组,每组8只,分别经尾静脉注入抗LOX-1抗体-NIR-797、非特异性IgG-NIR-797和单纯PBS。另取8只C57BL/6小鼠作为对照组,经尾静脉注入抗LOX-1抗体-NIR-797,注入探针24 h后取小鼠主动脉及主要脏器行NIRF成像,再取主动脉行油红O染色,比较NIRF成像面积和油红O染色面积比例的差异。结果:ApoE-/-小鼠经尾静脉注入抗LOX-1抗体-NIR-797探针24 h后,NIRF成像显示有强荧光信号聚集于小鼠离体主动脉的斑块区域,而注入非特异性IgG-NIR-797组斑块区信号明显减弱,PBS组及对照组C57BL/6小鼠仅见较弱荧光信号,单因素方差分析结果显示差异有统计学意义(F=232,P<0.001)。NIRF阳性面积与斑块油红O染色面积比例分别为(42.70±1.25)%和(42.78±1.51)%,两者差异无统计学意义(t=0.409,P=0.695)。结论:近红外荧光分子标记的抗LOX-1抗体探针能够特异性地显示ApoE-/-鼠主动脉粥样硬化斑块的分布,可以作为小鼠动脉粥样硬化斑块成像的有效工具,LOX-1将来可能成为一个新的评价动脉粥样硬化的良好靶点。
Objective: To explore the role of near-infrared fluorescence(NIRF) probes NIR-797 targeted anti-LOX-1 antibodies in atherosclerotic lesions imaging in aortic atherosclerosis lesions of ApoE deficient mice.Methods: Apolipoprotein E-/-(ApoE-/-) mice were fed with high cholesterol diet for 20 weeks to establish atherosclerotic model.Twenty-four ApoE-/-mice were randomly divided into 3 groups(8 in each group) and injected with anti-LOX-1-Ab-NIR-797,IgG-NIR-797 and phosphate buffered saline(PBS) respectively.Eight wild-type C57BL/6 mice were injected with anti-LOX-1-Ab-NIR-797 as control group.NIRF imaging of the aortas and major organs were performed 24 hours after tail vein injection of probes,followed by aortic excision for oil red O staining.Reults: NIRF imaging showed significant fluorochrome enhancement accumulation in atherosclerotic plaques in ApoE-/-mice with the anti-LOX-1-Ab-NIR-797 probe but not enough with the nonspecific IgG probe.Little signal enhancement was observed in ApoE-/-mice injected with the PBS and Wild-type C57BL/6 mice injected with anti-LOX-1-Ab-NIR-797 probe.The percentage of positive areas in imaging of NIRF and oil red O staining 24 h after injection of NIR-797 labeled anti-LOX-1 antibody were(42.70±1.25)% and(42.78±1.51)%.The difference between them was not statistically significant(t=0.409,P=0.695).Conclusion: This study demonstrates that anti-LOX-1-Ab-NIR-797 can be used in imaging atherosclerotic lesions.LOX-1 may be used as a target for molecular imaging of atherosclerotic plaque.
Objective: To explore the role of near-infrared fluorescence(NIRF) probes NIR-797 targeted anti-LOX-1 antibodies in atherosclerotic lesions imaging in aortic atherosclerosis lesions of ApoE deficient mice.Methods: Apolipoprotein E-/-(ApoE-/-) mice were fed with high cholesterol diet for 20 weeks to establish atherosclerotic model.Twenty-four ApoE-/-mice were randomly divided into 3 groups(8 in each group) and injected with anti-LOX-1-Ab-NIR-797,IgG-NIR-797 and phosphate buffered saline(PBS) respectively.Eight wild-type C57BL/6 mice were injected with anti-LOX-1-Ab-NIR-797 as control group.NIRF imaging of the aortas and major organs were performed 24 hours after tail vein injection of probes,followed by aortic excision for oil red O staining.Reults: NIRF imaging showed significant fluorochrome enhancement accumulation in atherosclerotic plaques in ApoE-/-mice with the anti-LOX-1-Ab-NIR-797 probe but not enough with the nonspecific IgG probe.Little signal enhancement was observed in ApoE-/-mice injected with the PBS and Wild-type C57BL/6 mice injected with anti-LOX-1-Ab-NIR-797 probe.The percentage of positive areas in imaging of NIRF and oil red O staining 24 h after injection of NIR-797 labeled anti-LOX-1 antibody were(42.70±1.25)% and(42.78±1.51)%.The difference between them was not statistically significant(t=0.409,P=0.695).Conclusion: This study demonstrates that anti-LOX-1-Ab-NIR-797 can be used in imaging atherosclerotic lesions.LOX-1 may be used as a target for molecular imaging of atherosclerotic plaque.
引文
[1]ROGER V L,GO A S,LLOYD-JONES D M,et al.Heart dis-ease and stroke statistics—2012 update:a report from the A-merican Heart Association[J].Circulation,2012,125(1):e2-e220.
[2]HE J,GU D,WU X,et al.Major causes of death among men andwomen in China[J].N Engl J Med,2005,353(11):1124-1134.
[3]VANCRAEYNEST D,PASQUET A,ROELANTS V,et al.Ima-ging the vulnerable plaque[J].J Am Coll Cardiol,2011,57(20):1961-1979.
[4]SANZ J,FAYAD Z A.Imaging of atherosclerotic cardiovasculardisease[J].Nature,2008,451(7181):953-957.
[5]MITRA S,GOYAL T,MEHTA J L.Oxidized LDL,LOX-1 andatherosclerosis[J].Cardiovasc Drugs Ther,2011,25(5):419-429.
[6]REISS A B,ANWAR K,WIRKOWSKI P.Lectin-like oxidizedlow density lipoprotein receptor 1(LOX-1)in atherogenesis:abrief review[J].Curr Med Chem,2009,16(21):2641-2652.
[7]BRILEY-SAEBO K C,NGUYEN T H,SAEBOE A M,et al.Invivo detection of oxidation-specific epitopes in atheroscleroticlesions using biocompatible manganese molecular magnetic im-aging probes[J].J Am Coll Cardiol,2012,59(6):616-626.
[8]CHEN I Y,WU J C.Cardiovascular molecular imaging:focus onclinical translation[J].Circulation,2011,123(4):425-443.
[9]NAHRENDORF M,ZHANG H,HEMBRADOR S,et al.Nano-particle PET-CT imaging of macrophages in inflammatory ath-erosclerosis[J].Circulation,2008,117(3):379-387.
[10]MORISHIGE K,KACHER D F,LIBBY P,et al.High-resolu-tion magnetic resonance imaging enhanced with superparamag-netic nanoparticles measures macrophage burden in atheroscle-rosis[J].Circulation,2010,122(17):1707-1715.
[11]BRILEY-SAEBO K C,CHO Y S,SHAW P X,et al.Targetediron oxide particles for in vivo magnetic resonance detectionof atherosclerotic lesions with antibodies directed to oxida-tion-specific epitopes[J].Journal of the American College ofCardiology,2011,57(3):337-347.
[12]WEN S,X X,LIU D F,et al.OxLDL-targeted iron oxide nan-oparticles for in vivo MRI detection of perivascular carotidcollar induced atherosclerotic lesions in ApoE-deficient mice[J].J Lipid Res,2012,53(5):829-838.
[13]卢瞳,文颂,周官辉,等.动脉粥样硬化斑块MRI和近红外分子影像的实验研究[J].中华核医学与分子影像杂志,2012,32(1):16-21.
[14]王圆圆,姚玉宇,张毅,等.近红外荧光分子标记的ox-LDL靶向小鼠动脉粥样硬化模型的成像研究[J].东南大学学报:医学版,2011,30(4):588-593.
[15]NEWBY A C.Metalloproteinase expression in monocytes andmacrophages and its relationship to atherosclerotic plaque in-stability[J].Arteriosclerosis Thrombosis and Vascular Biolo-gy,2008,28(12):2108-2120.
[16]RAZAVIAN M,TAVAKOLI S,ZHANG J,et al.Atherosclero-sis plaque heterogeneity and response to therapy detected byin vivo molecular imaging of matrix metalloproteinase activa-tion[J].J Nucl Med,2011,52(11):1795-1802.
[17]OHSHIMA S,FUJIMOTO S,PETROV A,et al.Effect of anantimicrobial agent on atherosclerotic plaques:assessment ofmetalloproteinase activity by molecular imaging[J].J AmColl Cardiol,2010,55(12):1240-1249.
[18]WALLIS de VRIES B M,HILLEBRANDS J L,van DAM G M,et al.Images in cardiovascular medicine multispectral near-in-frared fluorescence molecular imaging of matrix metalloprotei-nases in a human carotid plaque using a matrix-degradingmetalloproteinase-sensitive activatable fluorescent probe[J].Circulation,2009,119(20):e534-536.
[19]姚琼,邓钢,吴国球,等.MMP-9单克隆抗体超微超顺磁性氧化铁粒子的合成并行ApoE-/-小鼠动脉粥样硬化7.0TMR成像的研究[J].东南大学学报:医学版,2011,30(3):431-435.
[20]严雪娇,刘乃丰,吴国球.活体近红外成像技术在肿瘤研究中的应用[J].东南大学学报:医学版,2011,30(2):380-383.
[21]ISHINO S,MUKAI T,KUGE Y,et al.Targeting of lectinlikeoxidized low-density lipoprotein receptor 1(LOX-1)with99mTc-labeled anti-LOX-1 antibody:potential agent for ima-ging of vulnerable plaque[J].J Nucl Med,2008,49(10):1677-1685.
[22]LI D,PATEL A R,KLIBANOV A L,et al.Molecular imaging of atherosclerotic plaques targeted to oxidized LDL receptorLOX-1 by SPECT/CT and magnetic resonance[J].Circ Car-diovasc Imaging,2010,3(4):464-472.
[2]HE J,GU D,WU X,et al.Major causes of death among men andwomen in China[J].N Engl J Med,2005,353(11):1124-1134.
[3]VANCRAEYNEST D,PASQUET A,ROELANTS V,et al.Ima-ging the vulnerable plaque[J].J Am Coll Cardiol,2011,57(20):1961-1979.
[4]SANZ J,FAYAD Z A.Imaging of atherosclerotic cardiovasculardisease[J].Nature,2008,451(7181):953-957.
[5]MITRA S,GOYAL T,MEHTA J L.Oxidized LDL,LOX-1 andatherosclerosis[J].Cardiovasc Drugs Ther,2011,25(5):419-429.
[6]REISS A B,ANWAR K,WIRKOWSKI P.Lectin-like oxidizedlow density lipoprotein receptor 1(LOX-1)in atherogenesis:abrief review[J].Curr Med Chem,2009,16(21):2641-2652.
[7]BRILEY-SAEBO K C,NGUYEN T H,SAEBOE A M,et al.Invivo detection of oxidation-specific epitopes in atheroscleroticlesions using biocompatible manganese molecular magnetic im-aging probes[J].J Am Coll Cardiol,2012,59(6):616-626.
[8]CHEN I Y,WU J C.Cardiovascular molecular imaging:focus onclinical translation[J].Circulation,2011,123(4):425-443.
[9]NAHRENDORF M,ZHANG H,HEMBRADOR S,et al.Nano-particle PET-CT imaging of macrophages in inflammatory ath-erosclerosis[J].Circulation,2008,117(3):379-387.
[10]MORISHIGE K,KACHER D F,LIBBY P,et al.High-resolu-tion magnetic resonance imaging enhanced with superparamag-netic nanoparticles measures macrophage burden in atheroscle-rosis[J].Circulation,2010,122(17):1707-1715.
[11]BRILEY-SAEBO K C,CHO Y S,SHAW P X,et al.Targetediron oxide particles for in vivo magnetic resonance detectionof atherosclerotic lesions with antibodies directed to oxida-tion-specific epitopes[J].Journal of the American College ofCardiology,2011,57(3):337-347.
[12]WEN S,X X,LIU D F,et al.OxLDL-targeted iron oxide nan-oparticles for in vivo MRI detection of perivascular carotidcollar induced atherosclerotic lesions in ApoE-deficient mice[J].J Lipid Res,2012,53(5):829-838.
[13]卢瞳,文颂,周官辉,等.动脉粥样硬化斑块MRI和近红外分子影像的实验研究[J].中华核医学与分子影像杂志,2012,32(1):16-21.
[14]王圆圆,姚玉宇,张毅,等.近红外荧光分子标记的ox-LDL靶向小鼠动脉粥样硬化模型的成像研究[J].东南大学学报:医学版,2011,30(4):588-593.
[15]NEWBY A C.Metalloproteinase expression in monocytes andmacrophages and its relationship to atherosclerotic plaque in-stability[J].Arteriosclerosis Thrombosis and Vascular Biolo-gy,2008,28(12):2108-2120.
[16]RAZAVIAN M,TAVAKOLI S,ZHANG J,et al.Atherosclero-sis plaque heterogeneity and response to therapy detected byin vivo molecular imaging of matrix metalloproteinase activa-tion[J].J Nucl Med,2011,52(11):1795-1802.
[17]OHSHIMA S,FUJIMOTO S,PETROV A,et al.Effect of anantimicrobial agent on atherosclerotic plaques:assessment ofmetalloproteinase activity by molecular imaging[J].J AmColl Cardiol,2010,55(12):1240-1249.
[18]WALLIS de VRIES B M,HILLEBRANDS J L,van DAM G M,et al.Images in cardiovascular medicine multispectral near-in-frared fluorescence molecular imaging of matrix metalloprotei-nases in a human carotid plaque using a matrix-degradingmetalloproteinase-sensitive activatable fluorescent probe[J].Circulation,2009,119(20):e534-536.
[19]姚琼,邓钢,吴国球,等.MMP-9单克隆抗体超微超顺磁性氧化铁粒子的合成并行ApoE-/-小鼠动脉粥样硬化7.0TMR成像的研究[J].东南大学学报:医学版,2011,30(3):431-435.
[20]严雪娇,刘乃丰,吴国球.活体近红外成像技术在肿瘤研究中的应用[J].东南大学学报:医学版,2011,30(2):380-383.
[21]ISHINO S,MUKAI T,KUGE Y,et al.Targeting of lectinlikeoxidized low-density lipoprotein receptor 1(LOX-1)with99mTc-labeled anti-LOX-1 antibody:potential agent for ima-ging of vulnerable plaque[J].J Nucl Med,2008,49(10):1677-1685.
[22]LI D,PATEL A R,KLIBANOV A L,et al.Molecular imaging of atherosclerotic plaques targeted to oxidized LDL receptorLOX-1 by SPECT/CT and magnetic resonance[J].Circ Car-diovasc Imaging,2010,3(4):464-472.