血府逐瘀汤靶向“瘀闭玄府-钙调蛋白激酶紊乱网络”治疗创伤性脑损伤认知障碍的机制研究

Traumatic brain injury (TBI) with cognitive impairment is a clinically refractory disease, while traditional Chinese medicine exerts superior effects on cognitive improvement. Ancient doctors developed the Xuanfu theory from the meaning of "heart harbors veins, veins control spirit". The theory states that Xuanfu is the end of veins and the organization of spirit motion. Moreover, the theory could guide the treatment of cognitive impairment. Inspired by this theory, the pathogenesis of cognitive impairment post-TBI depends on the dysfunction of spirit motion caused by the Yubi Xuanfu. Xuefu Zhuyu Decoction possesses effects of “Hua Yubi and Tong Xuanfu”, and shows the clinical improvement of impaired cognition in TBI. Based on the previous studies and literature reports, the pathogenesis of Yubi Xuanfu can be interpreted by the disturbance of calcium-regulated protein kinases network. The efficacy of Xuefu Zhuyu Decoction is involved in the regulation of this network-related pathological process. According to this, we proposed a hypothesis that Xuefu Zhuyu Decoction improves the cognitive impairment post-TBI via targeting "Yubi Xuanfu syndrome relative network of calmodulin kinases". To verify this hypothesis, this project is going to perform PRM targeted quantitative proteomics coupled with physical behavior and molecular biology methods in rats with cognitive impairment after a controlled cortical impact injury. It will elucidate the modern mechanism of Yubi Xuanfu post-TBI, reveal the scientific connotation of Xuefu Zhuyu Decoction in treating cognitive impairment post-TBI from Yubi Xuanfu, and provide a new theoretical guidance for the treatment of TBI.

创伤性脑损伤（TBI）后认知功能障碍是临床难治病，而中医药在改善认知方面发挥优势疗效。历代医家从“心藏脉，脉舍神”发展出玄府理论，认为玄府是脉道终末端、运转神机结构，并临床指导认知障碍治疗。受此启发，TBI认知障碍病机在于瘀闭玄府，神机失运。血府逐瘀汤化瘀闭、通玄府，契合病机，临床改善TBI认知功能有效。基于既往工作基础及文献报道，瘀闭玄府病机可由钙调蛋白激酶紊乱网络诠释，血府逐瘀汤疗效涉及调控该网络相关病理过程，据此，提出血府逐瘀汤通过靶向调控“瘀闭玄府-钙调蛋白激酶紊乱网络”治疗TBI认知障碍的科学设想。本项目拟在中医玄府理论指导下，以控制性皮质损伤认知障碍大鼠为模型，运用靶向定量蛋白质组学联合在体行为学、分子生物学方法，验证该方调控钙调蛋白激酶网络的设想，发掘TBI后瘀闭玄府的现代机理，揭示血府逐瘀汤从瘀闭玄府论治TBI认知障碍的科学内涵，为中医药治疗TBI提供新的科学理论指导。

创伤性脑损伤（TBI）后认知功能障碍是临床难治病，而中医药在改善认知方面发挥优势疗效。历代医家从“心藏脉，脉舍神”发展出玄府理论，认为玄府是脉道终末端、运转神机结 构，并临床指导认知障碍治疗。受此启发，TBI认知障碍病机在于瘀闭玄府，神机失运。血府逐瘀汤化瘀闭、通玄府，契合病机，临床改善TBI认知功能有效。基于既往工作基础及文献报道，瘀闭玄府病机可由钙调蛋白激酶紊乱网络诠释，血府逐瘀汤疗效涉及调控该网络相关病理过程，据此，提出血府逐瘀汤通过靶向调控“瘀闭玄府-钙调蛋白激酶紊乱网络”治疗TBI认知障碍的科学设想。本项目在中医玄府理论指导下，以控制性皮质损伤认知障碍大鼠为模型，运用液相质谱对血府逐瘀汤进行了质量控制，运用水迷宫等行为学确定了血府逐瘀汤治疗TBI后认知障碍的最佳剂量为9g/kg，采用HE、尼氏、高尔基等染色从形态学上验证了血府逐瘀汤有效缓解了TBI后神经元的损失及突触丢失，最后联合靶向定量蛋白质组学、代谢组学、网络药理学等技术方法，证实了血府逐瘀汤对钙调蛋白激酶网络的多靶点效应（涉及Camk2b、 Camkk1、Camkk2、Dnajc5、Prkag2），验证了该网络核心病理机制-CaMKII介导的神经元突触再生，并论证了该病理过程与上游的谷氨酸神经递质释放机制相关。因此，本项目发掘了TBI后瘀闭玄府的现代机理，揭示了血府逐瘀汤从瘀闭玄府论治TBI认知障碍的科学内涵，为中医药治疗TBI提供了新的科学理论指导。