RBM3冷休克通路促进神经重塑的相关机制及其作为缺血性脑卒中后低温治疗新靶点的研究

血管再通是当前缺血性脑卒中的主要治疗方式，但再通后患者预后仍不理想。脑卒中早期是神经重塑窗口期，在此期间促进神经重塑更能改善功能康复。近期Nature研究显示RBM3介导冷休克通路与神经重塑密切相关。其是否参与卒中早期低温治疗的神经保护及神经重塑，改善功能是本研究的重点。我们预实验发现：卒中早期低温上调RBM3，促进神经功能恢复。据此我们提出研究假说：早期低温治疗促进卒中后神经重塑，RBM3介导冷休克通路发挥关键作用。本研究拟在体内、体外建立缺血再灌注模型，采用电镜、电生理、行为学和分子生物学技术探究RBM3介导冷休克通路如何改善神经重塑（神经元活性，神经功能和突触重塑）；用荧光素酶基因实验、特异性抑制剂、免疫共沉淀等技术明确HIF-1α/XBP1s转录复合物对FoxO1促进FAK磷酸化的影响是启动RBM3介导冷休克通路的分子机制。本研究将为早期再通基础上神经重塑和康复提供新方向和新理论。

Currently, revascularization is the main treatment for ischemic stroke. However，the paucity of patients reached by this therapy requires additional and supplementary anti-ischemic strategies. Immediately after stroke onset, there is a neuroplasticity window within which rehabilitation is especially effective. Recent studies have indicated that hypothermia-induced neuroplasticity involves the RBM3-mediated cold shock pathway, but it is unknown whether this pathway participates in neuroprotection and neuroplasticity in the context of hypothermia after stroke. Preliminary studies by our group have suggested that hypothermia administered early during cerebral ischemia increases RBM3 expression, and that this increased expression is associated with functional recovery. These findings led us to hypothesize that early hypothermia improves post-stroke neuroplasticity, and that this effect is mediated by the cold shock pathway. In this study, we will use electron microscopy, electrophysiology, and molecular biologic techniques in rodents and cell culture models to determine how neuroplasticity—as measured by assays of neuronal activity, neural function, and synaptogenesis—is improved through the cold shock pathway. Additionally, we will use the luciferase reporter gene assay, inhibitors, and co-immunoprecipitation to delineate the roles played by HIF-1α/XBP1 and FoxO1/FAK in initiating the RBM3-mediated cold shock pathway. Using these data, our study will provide evidence, for the first time, for the function of a novel mechanism supporting the neuroplasticity and rehabilitation achieved by early revascularization with hypothermia in the setting of ischemic stroke.