vWF-A2与波形蛋白结合导致微血栓形成在蛛网膜下腔出血后迟发性脑缺血中的作用及机制研究

蛛网膜下腔出血（SAH）后迟发性脑缺血（DCI）是导致结局不良的主要原因，脑微血栓形成是引发DCI的关键病理改变，但机制尚不明确且缺乏有效治疗手段。申请人前期研究发现，血管性血友病因子（vWF）作为反映凝血和血管内皮损伤的敏感性指标，可能参与了SAH后微血栓形成并有助于预测DCI发生。近期基础研究提示，vWF激活后可通过A2结构域与内皮细胞表面的波形蛋白结合。据此我们假设，SAH后可通过激活vWF-A2结构域与波形蛋白结合粘附于脑血管腔内并募集血小板，导致微血栓形成，进一步造成微循环灌注不足而发生DCI。本项目拟通过建立小鼠SAH模型，应用免疫组化、免疫荧光及ELISA等方法阐明vWF-A2结构域与波形蛋白结合在微血栓形成中的作用，通过激光散斑对比成像和动物行为学评价等技术验证其对脑微循环灌注和神经功能预后的影响，明确其导致SAH后DCI的机制，寻找关键的作用靶点，为治疗提供新的理论依据。

Delayed cerebral ischemia (DCI) after subarachnoid hemorrhage (SAH) is the main cause of poor outcome, and cerebral microthrombosis is the key pathological change that causes DCI, but the mechanism is not clear and there is a lack of effective treatment. Our studies have found that von Willebrand factor (vWF), as a sensitive indicator of coagulation and endothelial injury, may play an important role in microthrombosis formation after SAH and help to predict the occurrence of DCI. There has been evidence that the activated vWF can bind to vimentin on the surface of endothelial cells and platelets through the A2 domain. Therefore, we speculated that after SAH, vWF-A2 and vimentin interaction contributes to microthrombosis formation by adhere to the cerebrovascular lumen and recruit platelets, leads to insufficient microcirculation perfusion and DCI. To verify this hypothesis, we intend to investigate the role of vWF-A2/vimentin interaction on microthrombosis formation by establishing the SAH model in mice and using immunohistochemical, immunofluorescence and ELISA, further verify its effect on cerebral microcirculation perfusion and neurological function by laser speckle contrast imaging (LSCI) and neurobehavior tests, clarify the mechanism in the DCI after SAH, and find the key potential therapeutic targets to provide new ideas for the treatment, which has important research significance and clinical value.