雷洛昔芬、巴多昔芬抑制IL-6/gp130/STAT3信号途径及其抗炎、抗动脉粥样硬化作用的研究

Vascular inflammation is known to play a crucial role in the development of atherosclerosis, emphasizing an urgent need for anti-inflammatory treatment strategies. Proinflammatory cytokines, including interleukin-6 (IL-6) play very importatnt roles in atherosclerosis. It has been identified as an independent risk factor for CAD. IL-6 activates cells by first binding IL-6 receptor (IL-6R), which, after dimerization with gp130, leads to phosphorylation of Janus tyrosine kinases (JAKs). Then STAT3 is phosphorylated and activated. It is highly desirable to design and identify novel, small molecule drugs to target IL-6/ gp130/ STAT3, which might offer new options for anti-inflammatory treatment of atherosclerosis. Using a novel computational strategy for fragment-based drug design by combining MLSD (multiple ligand simultaneous docking) and drug repositioning, we demonstrated that Raloxifene and Bazedoxifene have a new function against IL-6/GP130 protein-protein interface in our previous study. .In the present study, we will detect the acivity of IL-6/gp130/JAK-STAT3 pathway in varies of cultrued vascular cells and also in atherosclerotic plaque from patients. We will examine the effects of Raloxifene and Bazedoxifene on VSMCs and endothelial cells proliferation and apoptosis. Ang II, TNF-α, oxLDL or LPS will be used to activate IL-6/gp130/JAK-STAT3 in endothelial cells, VSMCs and macrophages. Then the inhibitory effects of Raloxifene and Bazedoxifene on these cells will be tested in Vitro. Raloxifene and Bazedoxifene will also be further tested for efficacy in atherosclerosis in hypercholesterolemic rabbit and apoE-/- mice in vivo. The results will provide an innovative strategy for anti-inflammatory therapy by targeting IL-6/gp130/STAT3 signaling in atherosclerosis using Raloxifene or Bazedoxifene. These two drugs can also serve as lead compounds for optimization to speed up the development of novel drugs, which might be more potent and with desirable properties to disrupting IL-6/gp130/STAT3 for the treatment of CAD and other related chronic inflammatory disease.

动脉粥样硬化(AS)是一种血管慢性炎性疾病，目前尚无针对炎症反应的抗AS药物。炎症因子IL-6通过与配体IL-6R及gp130结合激活STAT3，在AS炎症反应中起着核心作用。我们以往的研究采用多配体对接技术，筛查发现两种雌激素受体调节剂雷洛昔芬、巴多昔芬能抑制IL-6与gp130结合，在肿瘤细胞能抑制IL-6诱导的STAT3磷酸化。但它们能否通过抑制IL-6与gp130结合，抑制血管炎症反应、防治AS尚不清楚。我们的预实验结果显示它们能抑制血管平滑肌细胞增殖、抑制炎症因子诱导的内皮及巨噬细胞STAT3磷酸化。本研究拟从细胞、动物水平，进一步检测这两种药物能否通过抑制IL-6/gp130/STAT3活性，抑制炎症介质诱导的内皮功能失调、平滑肌细胞增殖、巨噬细胞浸润、血小板聚集，从而抑制AS发生发展。为雷洛昔芬、巴多昔芬用于防治AS提供依据，并为开发新的抗炎性抗AS药物提供新方向和新的模板。

动脉粥样硬化（Atherosclerosis, AS）是一种血管慢性炎性疾病，目前尚无针对炎症反应的抗AS药物。研究证实，多种炎性标记物例如白介素-6（Interlukin-6）、肿瘤坏死因子-α（TNF-α）、白介素-1（IL-1）等都被证实存在于动脉粥样硬化斑块中，参与动脉粥样硬化的发生与发展，但其具体的机制尚不清楚。研究发现，IL-6信号通路是动脉粥样硬化炎性反应中的核心信号通路，与动脉粥样硬化关系最为密切。IL-6其主要作用是通过其下游信号分子信号传导与转录激活因子3（Signal Transducer And Activator of Transcription 3, STAT3）来实现。我们以往的研究采用多配体对接技术，筛查发现两种雌激素受体调节剂雷洛昔芬、巴多昔芬能抑制IL-6与gp130结合，在肿瘤细胞能抑制IL-6诱导的STAT3磷酸化。但它们能否通过抑制IL-6与gp130结合，抑制血管炎症反应、防治AS尚不清楚。.本研究拟从细胞、动物水平，进一步检测这两种药物能否通过抑制IL-6/gp130/STAT3活性，抑制炎症介质诱导的内皮功能失调、平滑肌细胞增殖、巨噬细胞浸润，从而抑制AS发生发展。为雷洛昔芬、巴多昔芬用于防治AS提供依据，并为开发新的抗炎性抗AS药物提供新方向和新的模板。.我们发现IL-6/gp130/STAT3信号途径在不同的血管壁细胞中分布和表达有所不同，而在炎症刺激下，血管壁细胞微环境改变，炎症因子浸润，血管内皮、平滑肌及巨噬细胞中持续异常激活STAT3信号通路，导致血管内皮细胞功能紊乱，进一步诱发血管平滑肌及巨噬细胞表现出过度的细胞活力和迁移能力。巴多昔芬和雷洛昔芬能够抑制炎症环境下血管壁细胞中IL-6/gp130/STAT3信号通路的激活，并调节血管壁内皮细胞功能，抑制血管平滑肌细胞及巨噬细胞的过度增殖和浸润。在在体实验中，通过构建高脂诱导的ApoE敲除小鼠动脉粥样硬化模型，我们发现炎症是高脂诱导动脉粥样硬化发生与发展的重要环节。在对血管组织进行检测中，我们发现高脂激活血管斑块组织中IL-6/gp130/STAT3信号途径，造成血管壁细胞功能紊乱，加重AS的发展。巴多昔芬和雷洛昔芬能够通过抑制IL-6/gp130/STAT3信号途径减轻高脂诱导的小鼠体内的炎症水平，改善血管壁细胞的功能，延缓AS的发生与发展。