携P-选择素抗体靶向微泡评价大白兔睾丸缺血/再灌注损伤的实验研究
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摘要
目的:探讨携P-选择素抗体靶向脂质微泡评价睾丸完全缺血/再灌注损伤的可行性。方法:制备普通脂质微泡(MB)和携P-选择素抗体靶向微泡(MBp)。30只大白兔随机分成对照组、缺血/再灌注损伤(IRI)0.5 h组、1 h组、2 h组、4 h组,每组6只,在缺血/再灌注术后行MB和MBp的双侧睾丸造影(间隔20 min),经过4～5 min待健侧睾丸实质中MB或MBp完全消失,测量术侧睾丸第一帧超声造影的声强度(F-P,10-5AU)。随后将术侧睾丸取下行免疫组化分析。结果:MBp造影图像显示各IRI组术侧睾丸F-P值分别大于各组的MB(P均<0.05),并在2 h[MBp(8.34±1.20)10-5AU,MB(1.87±0.25)10-5AU]最高。对照组术侧睾丸MB、MBp声强度未见明显差异(均为0 AU)(P>0.05)。免疫组化示对照组睾丸血管内皮无明显P-选择素表达,而IRI组P-选择素表达增加,并随时间延长而增多。结论:携P-选择素抗体靶向微泡超声造影能够评价睾丸IRI的炎症反应。
Objective: To explore the feasibility of evaluating complete ischemia-reperfusion injury( IRI) of the testis by contrast-enhanced ultrasonography with microbubbles( MB) targeted to P-selectin( MBp) in rabbits. Methods: We randomly divided 30healthy adult rabbits into five groups of equal number( control,0. 5 h IRI,1 h IRI,2 h IRI,and 4 h IRI),prepared phospholipid MB and MBp,and performed contrast-enhanced ultrasonography of the bilateral testes with MB or MBp at an interval of 20 min at different times after IRI. When MB or MBp disappeared completely in the healthy testis at 4 to 5 min after intravenous injection,we recorded the power of the first frame( F-P) in the IRI testes followed by immunohistochemical staining of the testis tissue. Results: CEU with MBp achieved a significantly higher F-P than that with MB in all the IRI groups( P < 0. 05),which was( 8. 34 ± 1. 20) versus( 1. 87± 0. 25) 10- 5AU at 2 hours,but there was no significant difference between MB and MBp in the control rabbits( 0 AU,P > 0. 05).Immunohistochemistry showed a significantly time-dependent increase in the expression of P-selectin in the vascular endothelial cells of the IRI testes,but not in those of the control. Conclusion: Contrast-enhanced ultrasonography with MBp can be used to evaluate the inflammatory reaction of testicular ischemia-reperfusion injury.
Objective: To explore the feasibility of evaluating complete ischemia-reperfusion injury( IRI) of the testis by contrast-enhanced ultrasonography with microbubbles( MB) targeted to P-selectin( MBp) in rabbits. Methods: We randomly divided 30healthy adult rabbits into five groups of equal number( control,0. 5 h IRI,1 h IRI,2 h IRI,and 4 h IRI),prepared phospholipid MB and MBp,and performed contrast-enhanced ultrasonography of the bilateral testes with MB or MBp at an interval of 20 min at different times after IRI. When MB or MBp disappeared completely in the healthy testis at 4 to 5 min after intravenous injection,we recorded the power of the first frame( F-P) in the IRI testes followed by immunohistochemical staining of the testis tissue. Results: CEU with MBp achieved a significantly higher F-P than that with MB in all the IRI groups( P < 0. 05),which was( 8. 34 ± 1. 20) versus( 1. 87± 0. 25) 10- 5AU at 2 hours,but there was no significant difference between MB and MBp in the control rabbits( 0 AU,P > 0. 05).Immunohistochemistry showed a significantly time-dependent increase in the expression of P-selectin in the vascular endothelial cells of the IRI testes,but not in those of the control. Conclusion: Contrast-enhanced ultrasonography with MBp can be used to evaluate the inflammatory reaction of testicular ischemia-reperfusion injury.
引文
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[12]陈少敏,杨莉,宾建平,等.携带抗P-选择素单抗靶向微泡和对比超声评价肾缺血再灌注损伤.中国医学影像技术,2008,24(7):985-988.
[13]李美瑜,纪丽景,杨莉,等.携抗P-选择素单抗靶向微泡在炎症模型微循环中黏附机制及行为方式的研究.中华超声影像学杂志,2010,19(6):532-535.
[14]王羽,薛恩生,张宇,等.超声造影在评价缺血后处理对兔睾丸完全扭转复位后保护效应中的应用.中华超声影像学杂志,2011,20(12):1074-1078.
[15]Lorant DE,Topham MK,Whatley RE,et al.Inflammatory roles of P-selectin.J Clin Invest,1993,92(2):559-570.
[16]Chukwuemeka AO,Brown KA,Venn GE,et al.Changes in Pselectin expression on cardiac microvessels in blood-perfused rat hearts subjected to ischemia-reperfusion.Ann Thorac Surg,2005,79(1)204-211.
[2]Turner TT,Tung KS,Tomosnasa H,et al.Acute testicular ischemia results in germ cell-specific apoptosis in the rat.Biol Reprod,1997,57(6):1267-1274.
[3]Zizzi HC,Zibari GB,Granger DN,et al.Quantification of P-selectin expression after renal ischemia and reperfusion.J Pediatr Surg,1997,32(7):1010-1013.
[4]Palazzo AJ,Jones SP,Anderson DC,et al.Coronary endothelial P-selectin in pathogenesis of myocardial ischemia-reperfusion injury.Am J Physiol,1998,275(5 Pt 2):H1865-H1872.
[5]Kaufmann BA,Lindner JR.Molecular imaging with targeted contrast ultrasound.Curr Opin Biotechnol,2007,18(1):11-16.
[6]Inaba Y,Lindner JR.Molecular imaging of disease with targeted contrast ultrasound imaging.Transl Res,2012,159(3):140-148.
[7]Schumann PA,Christiansen JP,Quigley RM,et al.Targetedmicrobubble binding selectively to GPIIb IIIa receptors of platelet thrombi.Invest Radiol,2002,37(11):587-593.
[8]Kaufmann BA.Lewis C,Xie A,et al.Detection of recent myocardialischaemia by molecular imaging of P-selectin with targeted contrast echocardiography.Eur Heart J,2007,28(16):2011-2017.
[9]Kaufmann BA,Carr CL,Belcik T,et al.Effect of acoustic power on in vivo molecular imaging with targeted microbubbles:Implications for low-mechanical index real-time imaging.J Am Soc Echocardiogr,2010,23(1):79-85.
[10]Lindner JR,Dayton PA,Coggins MP,et al.Noninvasive imaging of inflammation by ultrasound detection of phagocytosed microbubbles.Circulation,2000,102(5):531-538.
[11]纪丽景,杨莉,燕翼,等.P选择素靶向超声分子成像评价小鼠心肌缺血再灌注损伤.中华医学杂志,2009,89(24),1698-1701.
[12]陈少敏,杨莉,宾建平,等.携带抗P-选择素单抗靶向微泡和对比超声评价肾缺血再灌注损伤.中国医学影像技术,2008,24(7):985-988.
[13]李美瑜,纪丽景,杨莉,等.携抗P-选择素单抗靶向微泡在炎症模型微循环中黏附机制及行为方式的研究.中华超声影像学杂志,2010,19(6):532-535.
[14]王羽,薛恩生,张宇,等.超声造影在评价缺血后处理对兔睾丸完全扭转复位后保护效应中的应用.中华超声影像学杂志,2011,20(12):1074-1078.
[15]Lorant DE,Topham MK,Whatley RE,et al.Inflammatory roles of P-selectin.J Clin Invest,1993,92(2):559-570.
[16]Chukwuemeka AO,Brown KA,Venn GE,et al.Changes in Pselectin expression on cardiac microvessels in blood-perfused rat hearts subjected to ischemia-reperfusion.Ann Thorac Surg,2005,79(1)204-211.