芎芪有效配伍介导PKC调控紧密连接信号通路对溶栓后出血转化的作用与机制研究

Hemorrhagic transformation (HT) after thrombolysis is the most common serious complication after acute cerebral infarction. The tight junction signaling pathway of Claudin-5 / Occludin / ZO-1 mediated by PKC is to regulate blood brain barrier ( BBB ), which is an important scientific problem in studying the mechanism of HT after thrombolysis. We recently found that "effective compatibility of Xiongqi" can regulate BBB function and reduce HT after thrombolysis. But the damage of BBB and the PKC-mediated tight junction signaling pathway how to affect the HT after thrombolysis? "Xiongqi effective compatibility" how to reduce HT after thrombolysis,which through mediating the PKC-mediated tight junction signaling pathway. Those new issues remain unclear,but are the key questions. This subject intends to study the new target of HT after thrombolysis that affected by " effective compatibility of Xiongqi ", which based on PKC-mediated tight junction signaling pathway and through the body, outside experimental. ①Transmission electron microscope to observe the BBB structure ,and confirm the effect and mechanism of damage of BBB on HT after thrombolysis; ②To detect PKC, Claudin-5, Occludin and ZO-1, which are related to the tight junction signaling pathway ,and to clarify the effect and mechanism of " Xiongqi effective compatibility " on BBB; ③To observe the effect of "Xiongqixi effective compatibility" at different time points to prevent and control HT after thrombolysis, so as to provide a new theory for " Xiongqi effective compatibility" to reduce HT after thrombolysis.

溶栓后出血转化（HT）是急性脑梗死溶栓后最常见的严重并发症。PKC介导Claudin-5/Occludin/ZO-1紧密连接信号通路调控血脑屏障（BBB）是研究溶栓后HT机制的重要科学问题。我们近期发现“芎芪有效配伍”能调节BBB功能，减少溶栓后HT。但BBB损害及PKC介导紧密连接信号通路如何影响溶栓后HT？“芎芪有效配伍”怎样介导PKC调控紧密连接信号通路减少溶栓后HT？这些新的关键问题尚不清楚。课题拟用体内、外实验多方位基于PKC调控紧密连接信号通路研究“芎芪有效配伍”减少溶栓后HT的新靶点：①透射电子显微镜观察BBB结构，明确BBB损害对溶栓后HT的作用与机制；②检测紧密连接信号通路相关蛋白PKC、Claudin-5、Occludin、ZO-1，明确“芎芪有效配伍”对BBB的影响与机制；③观察“芎芪有效配伍”不同时点防治溶栓后HT的作用，为“芎芪有效配伍”减轻溶栓后HT提供新理论。

溶栓后出血转化（HT）是急性脑梗死溶栓后最常见的严重并发症。PKC是否介导Claudin-5/Occludin/ZO-1紧密连接信号通路影响血脑屏障（BBB）是研究溶栓后HT机制的重要科学问题。本研究通过构建SD大鼠自体血栓模型及SD大鼠脑微血管内皮细胞缺氧/再复氧模型，从体内外两方面证实了芎芪有效配伍能通过抑制PKCδ/MARCKS通路活性降低溶栓后MMP 9水平，并上调紧密连接蛋白Claudin-5/Occludin/ZO-1的表达，减少神经元细胞损伤，调节血脑屏障的通透性和完整性，以此发挥保护血脑屏障的关键作用，从而减轻溶栓后血脑屏障破坏，降低脑梗溶栓后的出血转化发生。通过验证PKC介导Claudin-5/Occludin/ZO-1紧密连接信号通路调控血脑屏障进而影响溶栓后出血转化，为出血转化的防治提供了研究思路与科学依据。同时，研究证实芎芪有效配伍介导PKC调控紧密连接信号通路为防治急性脑梗死溶栓后出血的新靶点，将有助于丰富中医药治疗脑病理论。此外，明确芎芪有效配伍抑制溶栓后出血损害的作用机制，为探索急性脑梗死溶栓后并发症治疗的新药物，以及为急性脑梗死更加安全有效的溶栓治疗提供一种新的治疗策略，在急性脑梗死的中医特色治疗方面具有重要临床意义。