针刺效应启动子研究--穴位局部ATP及其P2X3受体在针刺效应初始调控中的作用和机制研究

The purinergic signalling system is constituted by extracellular purine and pyrimidine signaling molecules and their receptors. Among these factors, the purine signaling molecule ATP and its P2X3 receptor play an important role in the transmission of nociceptive stimulus. Acupuncture, as is known, is a nociceptive stimulation, and researches have found the responsive changes of ATP at acupoints after acupuncture. Therefore, we can make the assumption that ATP and its P2X3 receptor at acupoints may take part in the process of the initial regulation of acupuncture. Acupuncture may increase the ATP release and activate the P2X3 receptor--On one hand, ATP as a neuron transmitter may deliver the acupuncture signal; on the other hand, acupuncture may cause the change of energy metabolism in the affected area, and the two may further stimulate acupuncture effect by affecting the action potential of afferent nerves. Based on the research platform of acupuncture causing viscera and systemic effect, this project plans to use multiple measures such as microdialysis, morphological methods, techniques of molecular biology and electrophysiology, and nonlinear dynamics analysis, to study the role of ATP and P2X3 receptor in the initial regulation of acupuncture effect, to explain the neurobiological mechanism of the primary response of acupuncture, to establish mathematical model, to reveal the key factors in the complex functional mechanism of acupuncture, and to provide the experimental basis for improving acupuncture effect.

细胞外的嘌呤和嘧啶信号分子及其作用受体构成嘌呤能信号转导系统，其中嘌呤信号分子ATP作用于P2X3受体在介导伤害性刺激中发挥着重要作用。针刺是一种伤害性刺激，已有研究表明穴位处ATP在针刺后会产生应答变化。我们认为，穴位局部的ATP及其P2X3受体参与构成了针刺效应启动子（即针效初始调控环节）：针刺可使穴位局部ATP释放增加并激活P2X3受体，一方面ATP作为神经递质传递针刺信息；另一方面引起局部能量代谢变化，二者相互作用，可通过影响支配穴位的传入神经动作电位，启动针刺效应信息。本研究在针刺引起脏腑效应和全身效应研究平台上，综合采用微透析、形态学、分子生物学、神经电生理技术和非线性动力学分析方法，研究ATP及P2X3受体在针刺效应初始调控中的作用，阐释其参与针刺效应初始调控的神经生物学机制，建立相关数学模型，揭示针刺起效的关键基础性因素，探索针效调控的新模式，为提高针刺疗效提供实验依据。

本项目将生理状态与病理状态相结合，在针刺调节胃功能的脏腑效应以及针刺全身镇痛效应多个研究平台上，综合采用高效液相、免疫组织化学染色、Western blotting、基因芯片等技术及生物信息学分析方法，研究穴位局部ATP等腺苷酸及其P2X3受体在针刺效应初始调控中的作用，初步明确了针刺复杂作用机制中的部分重要因素：一、证实了穴区ATP及其P2X3受体在针刺提高正常小鼠耐痛阈过程中具有重要作用；二、在针刺足三里改善大鼠胃运动低下的脏腑效应平台上，发现针刺可上调穴位局部P2X3受体表达，但对ATP等腺苷酸含量无明显影响；穴位局部P2X3受体可能在该针刺效应中起到一定作用；三、在针刺足三里对佐剂性关节炎大鼠镇痛效应的研究平台上，发现造模后穴位局部P2X3受体含量上调，针刺可以显著下调其含量，提示P2X3受体与炎性痛关系密切，针刺可能降低该受体表达而产生镇痛效应，但穴位局部P2X3受体是否参与针刺疼痛信号的初始传递过程仍需要进一步研究。