针刺加强药物入脑效应的腺苷2A受体调控机制研究

Acupuncture and medication is unique in traditional Chinese medicine (TCM) for the regulation of body function and disease prevention. It has been well acknowledged that acupuncture plays synergic and toxicity-attenuating effects on certain medicine, however, its mechanism is still not well defined. Our previous trials indicated acupuncture plus Aricept contributed to the improvement of cognitive function in patients. Considering the specialty of purines and activation mechanism of acupuncture, as well as the advances of blood brain barrier (BBB) regulation mediated by adenosine receptor, We propose a hypothesis that acupuncture may enhance the cerebral penetration of drugs via modulating the blood brain barrier (BBB) permeability mediated by adenosine receptor 2A. On this basis, we aim to evaluate the effects of cerebral penetration of drugs after acupuncture in the SAMP8 mice model base on the theory of “Du channel into brain”, through ethology, morphology, in vivo imaging and high performance liquid chromatography (HPLC), at the view of modulating aricept into brain and the concentration of acetyl choline. Moreover, we determined the effects of acupuncture on the BBB and adenosine receptor 2A signaling pathway using the target protein blockage in vivo, immunofluorescence, immunohistochemisty, Western Blot analysis and quantitative Real-Time PCR. Besides, we tried to discover the following cascade after acupuncture in enhancing the drug peneration into brain: acupuncture triggers elevation of adenosine and activation of A2A receptor, which subsequently triggered remodeling of cellular skeleton, reduced adhesion and tight junction, elevation of intercellular space, BBB opening, penetration of aricept into brain, and elevation of cognition. Our study provides evidence for the promotion of the acupuncture combined with medicine, and enriches the acupuncture research.

普遍认为，针灸对药物具有增效减毒效应，但其机制和规律研究处在起步阶段。本课题组临床证实针刺联合安理申能有效改善患者认知功能，结合嘌呤类物质与针灸作用机制的特殊性及腺苷2A受体调控血脑屏障的最新进展，我们提出“调控腺苷2A受体介导的血脑屏障通透性是针刺加强药物入脑的关键机制”假说，围绕该假说，在中医督脉入脑理论指导下，以SAMP8模型为载体，采用行为学、在体成像及高效液相技术，从调控安理申入脑，影响乙酰胆碱含量的角度，评价针刺加强药物入脑的效应；同时，结合靶蛋白在体阻断技术，运用免疫荧光、Western Blot、PCR等技术，检测针刺对血脑屏障和腺苷2A受体信号通路的影响作用，揭示针刺加强药物入脑的“针刺—腺苷含量提高，A2A受体激活—细胞骨架重塑、粘附降低、紧密连接减少—细胞间间隙增加，BBB开放—促进安理申入脑—增强改善认知能力”级联调控机制，为针药结合理论的建立提供证据支撑。

普遍认为，针灸对药物具有增效减毒效应，但其机制和规律研究处在起步阶段。本课题组临床证实针刺联合安理申能有效改善患者认知功能，结合嘌呤类物质与针灸作用机制的特殊性及腺苷2A受体调控血脑屏障的最新进展，我们提出“调控腺苷2A受体介导的血脑屏障通透性是针刺加强药物入脑的关键机制”假说，围绕该假说，在中医督脉入脑理论指导下，以SAMP8模型为载体，采用行为学、形态学、在体成像及高效液相技术，从调控安理申入脑，影响乙酰胆碱含量的角度，评价针刺加强药物入脑的效应；同时，结合靶蛋白在体阻断技术，运用免疫荧光、免疫组化、Western Blot、荧光定量PCR等分子生物学技术，检测针刺对血脑屏障和腺苷2A受体信号通路的影响作用，揭示针刺加强药物入脑的“针刺—腺苷含量提高，A2A受体激活—细胞骨架重塑、粘附降低、紧密连接减少—细胞间间隙增加，BBB开放—促进安理申入脑—增强改善认知能力”级联调控机制，为针药结合理论的建立推广提供证据支撑，丰富针灸学科学内涵。