靶向性声学造影剂与血管内皮细胞相互作用的实验研究

目的 制备携抗人VCAM-1单克隆抗体的白蛋白声学造影剂，观察其与损伤血管内皮细胞的相互作用，探讨评价血管内皮功能的新方法。方法 采用交联法将抗人VCAM-1单克隆抗体共价偶联到自制氟碳气体为核心的白蛋白微气泡表面，制备靶向性声学微气泡；倒置显微镜下分别观察普通白蛋白微气泡、靶向性微气泡与正常内皮细胞、损伤内皮细胞的结合作用，高倍视野下计数内皮细胞及黏附的微气泡的数目，通过计算微气泡与内皮细胞的比值对两者之间的结合作用进行定量分析。结果 无论是正常内皮细胞，或损伤内皮细胞，仅见少量的对照组微气泡的黏附作用；而镜下可见大量携VCAM-1单抗的白蛋白微气泡黏附在损伤内皮细胞表面，黏附数目显著高于黏附于正常内皮细胞表面的数目。结论 携VCAM-1单抗的靶向性声学造影剂能够特异性结合在损伤内皮细胞表面，开拓了超声成像技术检测血管内皮损伤、评价血管内皮功能新的研究领域。

Studies on Interactions Between Targeted Ultrasound Contrast Agents and Activated Vascular Endothelial Cells

Objective This study was performed to explore the interaction between the activated endothelial cells and the self-made targeted ultrasound agents,which were conjugated anti-human vascular cell adhesion molecule-1 (VCAM-1) monoclonal antibodies.Meanwhile,a new method of assessing endothelial function was further explored based on the above interactions.Methods Anti-VCAM-1 monoclonal antibodies were bound to the shells of the perfluocarbon gas-filled albumin microbubbles by covalently conjugation technology.The specific interaction between the microbubbles and the normal endothelial cells or activated(injured) endothelial cells was observed using inverted microscope,respectively.The microbubble adherence was quantified by calculating the ratio of the adherent microbubbles and endothelial cells.The results were compared with the control bubbles.Results There was rarely control microbubbles adherence to normal or activated endothelial cells.However,compared with normal endothelial cells,there were significantly more targeted microbubbles binding to the activated endothelial cells.Conclusions The ultrasound agents with anti-human VCAM-1 monoclonal antibody can specifically bind to the activated endothelial cells,which offered a new insight to the noninvasive detection of endothelial dysfunction using ultrasound imaging techniques.