以TNF-α和IL-1β为靶点超声分子成像及靶向治疗在动脉粥样硬化病变中的实验研究

Inflammation is involved in the development and progression of atherosclerosis (AS). As the key factor of acute cardiovascular events, it promotes vulnerable plaque rupture, thrombosis, and vascular occlusion .However, for vulnerable plaque is latent and has the feature of unexpected rupture, it is especially important for the prevention and treatment of atherosclerosis to take positive and effective anti-inflammatory measures and the early assessment of morphology and properties of atherosclerosis plaque. Tumor necrosis factor α(TNF-α) and interleukin-1β(IL-1β) are the two major initiating inflammatory cytokines in the inflammatory response, promoting the development of AS, vulnerable plaque formation and rupture. We will use APOE-/- mice undergoing ultrasonic microscopy, ultrasound microbubble molecular imaging targeted contrast agents for displaying AS plaque at the dynamic, non-invasive, and quantitative molecular level. Moreover, to evaluate the role and mechanism of TNF-α and IL-1β targeted therapy for AS , the molecular biology, histopathology and other experimental methods in vitro will be used. This study will elucidate the application prospects and molecular mechanism of TNF-α and IL-1β targeted ultrasound microbubble molecular imaging and therapy in the AS early warning and treatment, and to provide a new strategy and thinking for the early clinical diagnosis and treatment.

炎症贯穿整个动脉粥样硬化（AS）的发生发展过程，是促进AS进程、易损斑块形成及破裂、血栓形成和血管阻塞，引起急性心血管事件的关键因素。但是，由于易损斑块具有隐匿性和突发破裂的等特点，针对AS发生发展、易损斑块形成的早期预警及积极有效的抗炎治疗AS防治至关重要。肿瘤坏死因子α（TNF-α）和白细胞介素-1β（IL-1β）均是参与炎症反应的重要的始动炎症因子，是促进AS发生发展、易损斑块形成及破裂的关键分子。本申请拟使用APOE-/-小鼠，通过超声显微镜、超声微泡分子成像前沿技术在分子水平动态、无创显示并定量、定性AS斑块；采用分子生物学、病理组织学等实验方法结合体外分子研究探讨TNF-α和IL-1β靶向治疗对AS的作用及机制。本研究将借助以上实验方法，阐明以TNF-α和IL-1β为靶点超声分子成像及靶向治疗在AS早期预警和治疗中的应用前景及分子机制，为临床早期诊断和防治提供新的策略和思路。

炎症贯穿整个动脉粥样硬化（AS）的发生发展过程，是促进AS进程、易损斑块形成及破裂、血栓形成和血管阻塞，引起急性心血管事件的关键因素。但是，由于易损斑块具有隐匿性和突发破裂的等特点，针对AS发生发展、易损斑块形成的早期预警及积极有效的抗炎治疗AS防治至关重要。肿瘤坏死因子α（TNF-α）和白细胞介素-1β（IL-1β）均是参与炎症反应的重要的始动炎症因子，是促进AS发生发展、易损斑块形成及破裂的关键分子。本项目通过使用APOE-/-小鼠制备的AS模型以及TNF-α和IL-1β靶向的抑制剂—重组融合蛋白TFI，在体证实采用超声显微镜可进行早期AS斑块形成的活体观察，靶向UMI技术无创显示并定量、定性AS斑块，血清及主动脉组织检测结果证实AS发生发展会促进 TNF-α和IL-1β两种因子的表达，同时靶向抑制两种炎症因子后血清及主动脉组织中的炎症因子表达减轻，提示靶向双重抑制TNF-α和IL-1β会减轻AS炎症反应等损伤；在离体培养的人脐静脉内皮细胞上进行THP1-HUVECs 粘附实验，给予TFI干预会显著粘附的单核细胞数目。同时在体外分离培养的小鼠骨髓来源的巨噬细胞上，利用氧化型低密度脂蛋白刺激巨噬细胞，继续给予TFI处理，会明显降低细胞脂滴聚集。上述实验结果证实了以TNF-α和IL-1β为靶点通过抑制内皮细胞损伤减少单核细胞-内皮的粘附以及抑制巨噬细胞脂质功能，进而在AS血管损伤中发挥保护性调节的作用及机制，提示靶向针对TNF-α和IL-1β在AS相关疾病中具有较高的应用前景，为临床改善早期诊断和防治相关疾病的策略提供了新的依据和思路。