本实验室发现CRF可激活cPLA2而增加血管通透性。我们最新预实验显示小鼠脑缺血可激活Src-cPLA2通路，与报道的视网膜微血管内皮细胞低氧可致Src-PLD1-PKCϒ-cPLA2激活一致；CRF通过CRFR1在暂时性大脑中动脉阻塞模型（tMCAO）中降低血脑屏障通透性、减小水肿率、减小梗死面积；但在永久性大脑中动脉阻塞模型（pMCAO）则抵消S1PR2拮抗剂JTE013的脑血管保护作用。提示CRFR1对脑卒中的影响复杂、与缺血时间/再灌注损伤相关、且与S1PR2有密切关联。本项目将采用tPMCAO 与pMCAO，结合慢病毒载体注射探讨CRFR1对缺血性小鼠脑卒中的影响及作用机制；采用脑血管内皮细胞bEnd3 与hBMVEC结合慢病毒载体转染探索CRFR1对内皮单层通透性的影响及Src-PLD1-PKCϒ-cPLA2信号通路的参与。研究将为探索脑卒中发病机制及药物防治靶点提供新的视角。

Our previous results show that CRF can increase vascular permeability via activation of cPLA2. We recently found that experimental brain ischemia caused activation of Src-cPLA2, which is consistent with the report that in HRMVECs hypoxia induced activation of Src-PLD1-PKCϒ-cPLA2 pathway. Moreover, we found that CRFR1 plays a role of decreasing vascular permeability in transient MCAO (tMCAO). However, on the contrary, in permanent MCAO (pMCAO), CRFR1 activation offset the vasoprotective effect of S1PR2 antagonist, JTE013. These results suggest that the working mechanisms of CRFR1 in the two models are complex and different, which may be attributed to the different ischemic time & reperfusion injury and may be highly associated with S1PR2. The present project will further focus on the effect of CRFR1 and action mechanism in tMCAO and pMCAO, combining the method of CRFR1-interfering virus carrier injection. Furthermore, two brain vascular endothelial cells, bEnd3 and hBMVEC will be used for exploration of CRFR1 action and the participation of Src-PLD1-PKCϒ-cPLA2 pathway. This study will provide a new insight into the stroke mechanism and its therapeutic target.