基于PI3K/AKT信号转导通路探讨破血化瘀法对脑出血大鼠及体外神经细胞凋亡模型的干预机制

Cerebral hemorrhage is also named hemorrhagic stroke, treatment for acute phase of cerebral hemorrhage is an unsolved problem in medical domain. Traditional Chinese Medicine Master Ren ji-xue originated Poxuehuayu method for the disease. Our research group relied on series of research such as national research project and special scientific research, and we proved the advantage of Poxuehuayu method for cerebral hemorrhage and we also preliminarily confirmed its mechanism on molecular level is to affect cell apoptosis. On the basis of previous studies, this study is to investigate the mechanism of Poxuehuayu method (Didang Tang) for cerebral hemorrhage in molecular and gene level, and to clarify the intervention mechanism of Poxuehuayu method for reduction of cerebral hemorrhage rats' perihematoma tissue injury and neural cell apoptosis model in vitro on the basis of PI3K/Akt signal transduction pathway which is after NGF receptor, in the main pathway of mitochondrial mediated apoptosis, and in the main outcome measures of p-Akt, Bcl-2 protein family and caspase. To provide a reliable experimental basis for Poxuehuayu method for acute phase of cerebral hemorrhage and clarify its scientific connotation.

脑出血又称出血性中风，本病急性期的治疗是医学领域尚未解决的难题之一。国医大师任继学教授首创以破血化瘀法为主治疗本病。本课题组前期依托国家攻关课题、行业专项等系列课题，证实了破血化瘀法治疗脑出血的优势，同时在分子水平初步证实了本法主要作用机制为影响细胞凋亡。本课题拟在前期研究基础上，以NGF受体后PI3K/AKT信号转导通路为基础，以线粒体介导细胞凋亡为主要研究途径，以P-Akt、Bcl-2蛋白家族及caspase家族为主要观察指标，通过整体动物实验和体外神经细胞实验，在分子、基因水平探讨破血化瘀法（抵当汤）治疗脑出血的作用机制，阐明破血化瘀法减轻脑出血大鼠血肿周围组织损伤及在体外神经细胞凋亡模型中的干预机制，为破血化瘀法治疗脑出血急性期提供可靠的实验基础，并阐明其科学内涵。

脑出血又称出血性中风，本病急性期的治疗是医学领域尚未解决的难题之一。国医大师任继学教授首创以破血化瘀法为主治疗本病。本课题以PI3K/AKT信号转导通路为基础，以线粒体介导细胞凋亡为主要研究途径，通过动物实验和体外实验，在分子、基因水平探讨破血化瘀法（抵当汤）治疗脑出血的作用机制，阐明破血化瘀法减轻脑出血大鼠血肿周围组织损伤的干预机制。实验结果分三部分：脑出血动物实验表明抵当汤能够明显改善脑出血大鼠神经功能缺损；并能够激活 PI3K/AKT 信号转导通路调控下游凋亡相关蛋白，最终发挥抗凋亡作用；氯化铝诱导神经细胞凋亡模型研究结果表明，抵当汤通过激活NGF和TrkA从而激活PI3K/AKT 信号转导通路，进而调控线粒体凋亡相关蛋白的表达，抑制神经细胞凋亡；另外，缺血缺氧细胞模型结果表明，抵当汤通过调控GRP78-IRE1/PERK通路抑制内质网应激从而抑制线粒体凋亡。以上结果从动物实验和体外实验两方面验证，破血化瘀法通过调控线粒体凋亡途径，抑制脑出血导致的神经细胞凋亡，从而减轻脑出血大鼠血肿周围组织损伤，为临床应用该法治疗脑出血急性期提供可靠的实验数据支持。