以Ox-apoB特异性片段行免疫干预改善内皮细胞连接屏障及生存状态减轻动脉粥样硬化及相关损害研究

The endothelial dysfunction and endothelial cell survival state mediated by Ox-LDL infiltrating endothelial cells through lectin-like oxidized low desity lipoprotein receptor-1 (LOX-1) play a critical role in the pathogenesis of atherosclerosis and related lesions. However, there are still no favorable and-specific interventive methods to block this pathological process. Our preliminary study found that the antibodies against Ox-apoB specific peptide fragments related endothelial epitopes could improve Ox-LDL related endothelial cell junction barrier damage and reduce endothelial cell apoptosis through LOX-1 pathway, and actively immunize mice with this Ox-apoB specific peptide fragments can reduce the development of atherosclerosis. On these grounds, we hypothesized that the immunization treatment with Ox-apoB specific fragments could block Ox-apoB related endothelial damage and lipid deposition through LOX-1, and inhibit the development of atherosclerosis and related lesions. Therefore, this study is intended to protect endothelial cell barrier junction, prevent endothelial cell pyroptosis and apoptosis, improve endothelial cell survival state by active and passive immunization strategies with this Ox-apoB specific fragments and observe protective effect on the atherosclerosis plaque formation and related lesions. And we will use LOX-1 knockout mice to verify this process involved LOX-1 pathway; at the same time, the above effects and mechanisms will be verified in endothelial cells in vitro. And this study also tries to investigate the effects of immunization strategiest on the pathogenesis of neoatherosclerosis. Finally, the main goal of this study is to explore the feasible immune intervention way to prevent the pathogenesis of atherosclerosis and related lesions through protection mechanisms of the endothelial cells.

Ox-LDL通过LOX-1浸润内皮细胞所致内皮功能障碍和内皮细胞生存危害对动脉粥样硬化的发展至关重要，但现仍无良好的特异性干预手段阻滞这一过程。我们的初步研究发现针对Ox-apoB特异性内皮粘附抗原表位的抗体可改善Ox-LDL通过LOX-1对内皮屏障所致的损害并减少内皮细胞凋亡；且主动免疫小鼠此片段可减轻粥样硬化的发生。据此我们推测以Ox-apoB特异性片段行免疫干预可阻滞Ox-apoB通过LOX-1引起的内皮损伤和脂质沉积，抑制粥样硬化及相关损害。据此，本研究拟采用主动及被动免疫策略观察动物内皮细胞屏障连接、焦亡及凋亡等生存状态及斑块形成的保护作用，并以LOX-1敲除鼠验证LOX-1介导此致病通路；同时，对上述效应及作用机制在离体内皮细胞上进行系列验证；本研究还试图借此探讨此免疫策略对支架内新生动脉粥样硬化有无作用。最终从内皮细胞保护的角度为动脉粥样硬化及相关损害提供可行的免疫干预途径。

Ox-LDL通过LOX-1浸润内皮细胞所致内皮功能障碍和内皮细胞生存危害对动脉粥样硬化的发展至关重要，但现仍无良好的特异性干预手段阻滞这一过程。我们的初步研究发现针对Ox-apoB特异性内皮粘附抗原表位的抗体可改善Ox-LDL通过LOX-1对内皮屏障所致的损害并减少内皮细胞凋亡；且主动免疫小鼠此片段可减轻粥样硬化的发生。我们的研究聚焦于动脉粥样硬化发生的初始环节，即氧化低密度脂蛋白对血管内皮细胞的破坏。血管内皮细胞通过其LOX-1受体与循环中的氧化低密度脂蛋白结合后诱发一系列炎症反应，内皮屏障的破坏最终导致级联反应。大量的低密度脂蛋白及氧化低密度脂蛋白通过血管内皮屏障，最终导致不可逆的动脉粥样硬化斑块。据此我们课题核心思路以Ox-apoB特异性抗体免疫干预阻断Ox-LDL/LOX-1通路，从而减轻LOX-1途径引起的内皮损伤和内膜下脂质沉积，抑制粥样硬化对内皮细胞的损害及相关损害。本研究采用主动及被动免疫策略观察动物内皮细胞屏障连接、焦亡及凋亡等生存状态及斑块形成的保护作用；同时，对上述效应及作用机制在离体内皮细胞上进行系列验证。我们的研究表明，Ox-LDL可通过内皮细胞LOX-1受体进入细胞，引起血管内皮细胞功能性损伤及细胞死亡。血管内皮屏障的破坏导致动脉粥样硬化快速进展。抗体阻断阻断ox-LDL/lox-1受体通路的结合，通过保护内皮屏障完整性可有效抑制动脉粥样硬化。我们研究从内皮细胞屏障保护的角度为动脉粥样硬化及相关损害提供理论依据，同时提出可行的免疫干预途径，具有较好的创新性及临床应用价值。