靶向性声学造影剂与不同程度损伤血管内皮细胞相互作用的实验研究  

Studies on interactions between targeted ultrasound contrast agents and damaged vascular endothelial cells in different degrees

作　　者：刘娅妮[1] 邓又斌[1] 刘冰冰[1] 杨好意[1] 毕小军[1] 张清阳[1] 王红英[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院超声影像科,湖北武汉430030

出　　处：《中国医学影像技术》2006年第11期1617-1620,共4页Chinese Journal of Medical Imaging Technology

摘　　要：目的观察携VCAM-1单克隆抗体的白蛋白声学造影剂与不同程度损伤血管内皮细胞的相互作用,并探讨这种靶向性声学微气泡对内皮细胞的黏附作用与内皮细胞受损程度之间的关系。方法以不同浓度肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)作为干预因子培养6组不同程度损伤的人脐静脉血管内皮细胞,采用免疫细胞化学染色与病理图文分析法检测各组血管内皮细胞表面VCAM-1的表达水平;采用SPDP交联法制备携VCAM-1单克隆抗体的靶向性白蛋白微气泡,倒置显微镜下观察其与各组损伤内皮细胞的结合作用,并于高倍视野下(200×)计数内皮细胞及黏附的微气泡的数目,计算微气泡与内皮细胞的比值作为微气泡的黏附率;各组损伤内皮的免疫微气泡的黏附率与其表面VCAM-1的表达水平进行相关性分析。结果随着TNF-α干预浓度的增高,内皮细胞表面VCAM-1的表达水平逐渐增高,且各组之间具有显著性差异;镜下观察可见损伤内皮细胞免疫微气泡黏附作用亦随着干预浓度的增高而增强,且微气泡对损伤内皮细胞的黏附率与细胞表面VCAM-1的表达水平呈正相关。结论携VCAM-1单克隆抗体的靶向性声学造影剂不仅能够与损伤血管内皮细胞特异性结合,其黏附率亦能够反映内皮细胞损伤的程度。Objective To detect the interactions between the graded damaged endothelial cells and the self-made targeted ultrasound agents, which were conjugated anti-human vascular cell adhesion molecule-1 （VCAM-1） monoclonal antibodies. Methods Human umbilical vein vascular cells were stimulated with TNF-α at the indicated concentration （0-300 U/ml） for the same time （24 h）. There were six groups of endothelial cells which were damaged differently in degree. Cell surface expressions of VCAM-1 were detect by immunocytochemistry. Monoclonal antibodies to VCAM-1 were bound to the shells of the perfluocarbon gas-filled albumin microbubbles by SPDP conjugation technology. The specific interactions between these microbubbles and each group of endothelial cells were observed using inverted microscope. The microbubbles adherence was quantified by calculating the ratio of the adherent microbubbles and endothelial cells. The results were compared between ev- ery two groups. And correlation analysis was performed between the targeted microbubbles＇ adherence and the cell surface expression of VCAM-1. Results After TNF-α stimulation, the expressions of VCAM-1 in groups of damaged endothelial cells were obviously increased. And the expressions were dose-dependent. With the increase of TNF-α stimulation concentration, there were more and more targeted microbubbles adherent to the corresponding endothelial cells. And the correlation analysis showed there was positive correlation between the targeted microbubbles＇ adherence and the cell surface expression of VCAM-1. Conclusion The ultrasound agents with anti-human VCAM-1 monoclonal antibody can specifically bind to the activated endothelial cells. And these ultrasound agents＂ adherence provides a new way to detect the damaged degrees of endothelial cells.

关 键 词：抗体 超声检查 造影剂 血管细胞黏附分子-1 肿瘤坏死因子-α 血管内皮细胞 

分 类 号：R-33[医药卫生] R445.1