基于新型三靶向微泡的超声分子影像早期诊断动脉粥样硬化及其疗效评价研究

Atherosclerosis (AS) is a chronic and inflammatory disease that poses serious threat to human health. Due to the insidious nature of disease development, early diagnosis is still very challenging with currently available technology. With the advancement of ultrasonic molecular imaging technology, it is more feasible now to characterize the initial inflammatory changes of AS for early diagnosis. In this project, we plan to take advantage of a new technology by targeting microbubbles to SLeX, P-selectin and VCAM-1. These molecules in turn can specifically bind to initial inflammation cytokines, and evaluate their morphological characteristics and adhesive capacity both in vivo and vitro. We will also utilize contrast-enhanced molecular ultrasound in a mouse (DKO) model of early AS to observe the imaging conditions at different time points. By combining ultra-frequency ultrasound with histology, immunohistochemistry, and western blot, we plan to evaluate whether we can overcome the past restrictions to carry out molecular imaging in the artery for early detection of AS. Furthermore, in order to investigate whether statins, the best anti-lipidemic agents for AS treatment, can exert their anti-inflammatory effects on early stage AS, we will assess the graded response of their therapeutic effects on the above mouse model. For this purpose, we will feed the mice with low, medium and high dose of atorvastatin and general diet for subgroup observation. These studies will allow us to potentially identify a novel strategy for effective treatment of inflammation, reliable means of risk assessment, and accurate prognosis for patients with early stage AS.

动脉粥样硬化（AS）是一种慢性炎症性疾病，严重威胁人类健康。由于发病隐匿，现有检测方法难以实现其早期诊断。随着超声分子影像的发展，可对AS早期炎症过程进行分子水平成像，从而使其早期诊断成为可能。本项目拟制备同时携快速结合型和稳定结合型配体（VCAM-1/ICAM-1/P-selectin）的新型三靶向超声微泡，通过表征检测、体外寻靶、平行板流动腔以及早期AS小鼠体内的超声分子靶向实验，探究其能否克服以往局限，提高微泡在高流速、高剪切力动脉血管中的靶向黏附性能。同时，使用不同剂量阿托伐他汀喂养早期AS小鼠模型，通过超声分子靶向显像方法多时间点观察他汀类药物对AS内皮炎症因子干预的靶向成像效果，再结合组织学、免疫组织化学、蛋白印迹等病理学方法，探究他汀类药物能否发挥其抗炎作用，实现对早期AS内皮炎症的有效治疗，从而为临床制定出包括有效炎症干预和可靠风险评估手段的AS早期治疗策略奠定实验基础。

动脉粥样硬化(atherosclerosis，AS)导致的心血管疾病严重威胁人类健康，其发生发展过程均与持续的炎症反应相关，因此，监测炎症相关的病理变化对于动脉粥样硬化的早期诊断和疗效评价具有重要意义。超声分子成像作为一种有前景的非侵入性成像技术已被证明可用于评估动脉粥样硬化的程度，然而动脉血管因血流速度快、剪切应力大，AS的超声分子成像仍存在靶向超声微泡黏附及保留困难的问题。因此，本研究制备一种携VCAM-1单抗、ICAM-1单抗和Sialyl-LewisX聚合物的基于VCAM-1/ICAM-1/selectin三靶向超声微泡，提高超声微泡对早期AS的靶向及显像能力，通过超声分子影像及病理学等方法、于体内外早期AS模型中，探索其对AS的早期诊断能力并评价其能否应用于阿托伐他汀治疗早期AS的疗效评价。主要研究内容包括：（1）研究基于VCAM-1/ICAM-1/P-selectin三靶向超声微泡的制备方法及特征，使其具有良好的声学反应性能和靶向黏附性能。（2）研究该三靶向超声微泡的体外靶向结合能力及体内、外靶向超声成像效果，探究其能否实现小鼠AS的早期诊断。（3）研究该三靶向超声微泡的超声分子成像能否实现小鼠体内他汀类药物治疗早期AS效果的评价。. 本项目围绕新型超声造影剂的制备以及靶向超声造影剂对动脉AS的早期诊断及疗效评价的相关问题进行了深入的研究，取得的主要成果包括：（1）成功制备出携VCAM-1单抗、ICAM-1单抗和Sialyl-LewisX聚合物的三靶向超声微泡造影剂，并通过多种方法检测其表征及性能，证实该微泡可用于后续实验研究。（2）通过微泡表面三种配体功能的合理整合，体内及体外实验均证实该微泡主动靶向性能及成像效果的提高。（3）应用该三靶向超声微泡实现药物治疗对早期AS的治疗效果的超声分子成像评价，验证了阿托伐他汀对AS早期炎症的治疗效果。