Hippo-YAP信号通路调控紧密连接蛋白参与蛛网膜下腔出血后血脑屏障损伤的作用及机制研究

项目名称： Hippo-YAP信号通路调控紧密连接蛋白参与蛛网膜下腔出血后血脑屏障损伤的作用及机制研究

项目编号： No.81501002

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 陈渝杰

作者单位： 中国人民解放军第三军医大学

项目金额： 17.5万元

中文摘要： 血脑屏障(BBB)损伤是蛛网膜下腔出血(SAH)后早期脑损伤主要病理生理改变之一，其中针对紧密连接蛋白这一关键因素的调控，可能是促进BBB修复最具前景的策略。前期研究证实：SAH后内皮细胞β-Catenin 核转位受抑制、紧密连接蛋白表达降低，而Hippo-YAP通路关键蛋白MST1显著升高。近期文献亦提示：该通路可能参与SAH后BBB中内皮细胞的损伤，其中YAP能够抑制β-Catenin 下游转录，其机制是通过YAP与β-Catenin形成复合体，减少β-Catenin 核转位。据此我们推测：Hippo-YAP通路可能通过抑制内皮细胞β-Catenin 核转位及紧密连接蛋白表达参与SAH后BBB损伤。本项目拟以SAH救治新靶点——BBB损伤为切入点，建立在体和离体模型，结合分子生物学、双光子显微成像等技术，探讨Hippo-YAP通路的作用及机制，提供有助于SAH患者临床救治的实验依据。

中文关键词： 蛛网膜下腔出血；Hippo；血脑屏障；紧密连接蛋白；β-Catenin

英文摘要： Subarachnoid hemorrhage (SAH) is a deadly cerebrovascular disease with high morbidity and mortality. In recent years，research interest has been shifted to the early brain injury (first 72h) evoked by SAH. However, relative clinical trials cannot be proved effective so far, because they merely targeting injured neurons after SAH. These disappointing results might remind us that we should pay more attention into the widespread contacts among all intracranial cell types, as known as the Vascular Neural Network. The blood brain barrier is the most typical neurovascular unit in this vascular neural network, and also blood brain barrier damage is thought to be one of the main pathophysiological processes in SAH patients. In this case, repairing blood brain barrier has been considered as one of the most promising therapy strategies in rescuing and repairing brain injury after SAH. Thus, as the key factor of blood brain barrier structural proteins, the tight junction proteins become the high interested repairing target. Our previous study demonstrated thatβ-catenin nucleic translocation in cerebral endothelial cells was reduced after SAH, leading to reduced tight junction protein levels. Meanwhile, a key protein of Hippo-YAP pathway, MST1, significantly increased at 24 hours after SAH. Furthermore, recent studies indicated that Hippo-YAP pathway might involve in the blood brain barrier injury after SAH. Another study showed that YAP could inhibit the downstream transcription of β-catenin by combined withβ-catenin in cytoplasm. Therefore, based on current evidences, we purpose that Hippo-YAP pathway could inhibit tight junction protein transcription by reducing β-catenin nucleic translocation, resulting in blood brain barrier injuried, which finally leads to neurological deficits after SAH. In the current project, by using two photon microscopy, immunofluorescence, laser scanning confocal microscope, behavior evaluation, cellular and molecular biology techniques，we will document the role of Hippo-YAP pathway in blood brain barrier injury after SAH, and then, identify the mechanisms of how Hippo-YAP pathway inhibiting tight junction proteins transcription. We hope that the current project could clarify the role and mechanism of Hippo-YAP pathway involving in blood brain barrier injury after SAH, and finally provide an innovational strategy to prevent early brain injury and improve clinic outcome in SAH patients.

英文关键词： Subarachnoid Hemorrhage;Hippo;Blood Brain Barrier;Tight Junction Protein;β-Catenin