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

目的 荧光显微镜直视下对比评价携抗P-选择素单抗靶向微泡与同型对照微泡在微循环中的黏附机制及行为方式.方法 构建携荧光FITC的抗P-选择素单抗靶向微泡(MBp)和同型对照微泡(MBiso),并随机经静脉注入小鼠提睾肌炎症模型.20倍荧光显微镜直视下观察并记录5min内两种微泡在提睾肌微循环中的黏附情况,并对不同黏附方式的微泡进行计数.应用image-pro-plus分析软件对微泡进行定点追踪,并对其黏附过程中速度的变化进行定量测定.结果 荧光显微镜下观察可见MBp组与内皮黏附数量高达(8.4±2.1)个/视野,MBiso组仅为(0.8±0.8)个/视野,两者间差异有统计学意义(P＜0.01);MBp和MBiso组白细胞黏附数量分别为(3.6±0.6)个/视野、(2.2±0.8)个/视野,两者间差异无统计学意义(P＞0.05).微泡黏附过程速度变化曲线显示MBp和MBiso分别以流动速度逐渐和迅速降低两种方式实现对靶组织的黏附.结论 MBp和MBiso具有不同的黏附方式,MBp能更高效、特异地黏附于炎症组织血管内皮上,为评价血管内皮炎症反应或其他组织损伤的应用提供了理论基础.

Assessment of adhesion mechanisms and behaviors of P-selectin targeted microbubbles in inflammatory microcirculation model

Objective To evaluate the adhesion mechanisms and microvascular behaviors of P-selectin targeted microbubbles and isotype control microbubbles.Methods The targeted microbubbles with FITC fluorescence and antibodies of P-selectin (MBp), and the isotype control microbubbles with FITC fluorescence and isotype antibodies(MBiso) were constructed.MBp and MBiso were injected intravenously in random to wild-type mice with TNF-α stimulus.The mechanisms and behaviors of MBp and MBiso in cremasteric venules were assessed by fluorescence microscopy.And then velocity variation of ultrasound microbubbles in microvascular was analyzed by tracking fixed-point technology of image-pro-plus software.Results The retention of microbubbles directly to venular endothelium amounted to (8.4 ± 2.1 )/view in MBp-group,while it was only (0.8 ± 0.8)/view in MBiso-group.Difference was evident between of the two groups( P ＜0.01 ).The numbers of MBp and MBiso attachment to activated leukocytes were (3.6± 0.6)/view and (2.2± 0.8)/view separately.There was no obvious difference between the two groups( P ＞0.05).The variation curves of velocity in MBp and MBiso targeting to venular endothelium were different.The flowing rate decreased gradually in MBp-group,while it decreased rapidly in MBiso-group.Conclusions MBp and MBiso have different behaviors targeting to venular endothelium, MBp could adhere to vascular endothelial efficiently and specifically, these results suggest MBp may be used to assess inflammation of vascular endothelial or other tissue injury effectively.