基于外泌体的功能特点以及胶质—神经细胞的相互作用关系探讨加味逍遥散治疗肝郁脾虚型肝癌的效应机制

Recent research data indicated that exosome is related to the Chinese medical theory of Zangxiang; epidemiological investigation showed that about half of the HCC have the complication of depression, and depression is also an independent etiological factor of HCC; former research data suggested that Xiaoyao-San not only treated depression but also treated HCC. The former NSFC Fund indicated that exosome play an important role in treating HCC and play an important role in the interaction of glia and neurons. And then, under the foundation of the above researches, this project submits the hypothesis: the mechanism of Jiawei Xiaoyao-San on treating HCC maybe via the effect of exosome on the hepatic cell and on the interaction of glia and neurons, and then target the central neurons and liver. To clarify the mechanism, this project will collect the patients with HCC from the hospitals, and figure out the effect of Jiawei Xiaoyao-San on secretion of exosome and the effect on influence of the RNAs and protein of exosome; this project also use in vitro experiments to figure out whether the treated exosome by Jiawei Xiaoyao-San has the effect on influence hepatic cells and/or the interaction of glia and neurons or not? This project not only play an important role in clarify the mechanism of Jiawei Xiaoyao-San but also provided new proof for the demonstration of Chinese medical theory of Zangxiang.

最新研究提示，外泌体与中医臧象理论相关；流行病学调查发现，原发性肝癌（HCC）一半左右并发抑郁症，而抑郁症也是HCC诱因之一；前期研究结果显示，加味逍遥散不但治疗抑郁症，对HCC也具治疗作用。前期参加和主持的国基金项目也提示，外泌体在HCC和胶质-神经相互作用调节中具有重要作用。由此，本项目提出工作假说：加味逍遥散对肝郁脾虚型肝癌的治疗，可能是通过外泌体分别调节肝功能和胶质-神经细胞（相互作用），从而达到情志和肝脏的双重治疗。围绕假说，本项目拟通过临床研究，结合蛋白质组学和基因测序方法，明确加味逍遥散对肝郁脾虚型肝癌的治疗是否与外泌体分泌量和外泌体内蛋白、RNAs有关？同时结合体外实验，观察经过加味逍遥散干预的外泌体是否对肝癌，胶质-神经系统非接触式共培养系统具有干预和调节作用？本项目研究不但对阐释加味逍遥散抗HCC作用机制具有重要意义，还有助于为中医臧象理论科学内涵研究提供新的实验依据。

中医藏象理论提示，肝系统不仅包括肝组织也包括肝功能状态，包括气机的疏通，情志的舒畅等。临床上加味逍遥散对肝癌及肝癌引起的抑郁的治疗作用，正体现其作用于肝藏象的功能特点。本项目通过对加味逍遥散的蛋白质组学和代谢组学及神经功能的检测发现，加味逍遥具有抗癌、调节免疫功能，改善情志等网状功能和多重靶点作用，进一步通过对外泌体miRNAs测序和外泌体与神经-胶质的相互作用关系研究发现，加味逍遥散的多重网络靶点和效应与调节外泌体及外泌体中miRNAs有关，而肝癌来源外泌体能损害神经功能和神经突起的发育，加味逍遥散调解后的外泌体则能改善肝癌外泌体导致的神经损伤。由此推断，加味逍遥散具有通过外泌体miRNA发挥双重作用：一方面通过外泌体miRNAs作用于机体，调节血清蛋白质和代谢物，发挥抗肝癌作用，一方面通过外泌体中miRNAs进入中枢神经系统调节神经突起的发育，影响神经兴奋性。该项目的研究为加味逍遥散辅助治疗肝癌及肝癌并发症抑郁的有效性提供了科学依据，同时也为中医藏象理论的物质基础研究提供了思路。