肾及其原、络、俞、募穴相关神经元的形态学和化学特征

The correlation of the acupoint and viscera is the basis and core of the theroy of meridan, and also is one of mainlines on acupuncture research in recent several decades. Although the previous studies prove that nervous system has closely relation with acupoint and viscera from different point of view, due to the techenical limitation of early times, untill now, there are absent of systematic morphological study on this field. Recently, following the rapid developing of neuroanatomical techniques, under the new condition, it becomes possible to further study the correlation of the acupoint and viscera under the direction of the functional related various neurons with morphological approach. In this study, we chose kidney and its associated Yuan-point: Taixi, Luo-point: Dazhong, back-shu-poin: kidney-shu and Mu-point: Jingmen as the breakthrough points, and use new high-sensitive neural tracing technique and fluorcent immunohistochemical technique, and combine with laser confocal microscopic technique to further investigate the morphological and chemical characteristics of neurons associated kidney and its related Yuan-, Luo-, Shu- and Mu-point. Through observing the regular innervation, morphological and chemical characteristics of afferent and efferent projected neurons to kidney and its related specific points, we may reveal the multiple correlation of the kidney and its related Yuan-, Luo-, Shu- and Mu-point with the sensory, motor and autonomic nervous systems on the cellular level. Thus, we believe that this project could not only provide neuroanatomical basis for further explaining the underlying mechenism of neuroscience on the correlation of the acupoint and viscera,but also guide to select specific acupionts for treating correlated visceral diseases.

经穴与脏腑相关性是经络理论的基础和核心内容，多年来一直是针灸学研究的主线之一。虽然以往的研究从不同的角度显示经穴和脏腑相关性与神经系统存在着紧密的联系，但是由于受到技术条件的限制，到目前为止，在这一领域还缺少系统性的形态学研究。近年来神经解剖学技术快速发展，为从形态学入手深入研究多种神经元参与下的经穴和脏腑联系创造了新的条件。本研究以肾及其原穴-太溪、络穴-大钟、背俞穴-肾俞和募穴-京门为切入点，综合运用新一代高敏感神经示踪和免疫荧光组织化学技术并结合共聚焦显微镜技术深入研究肾及其原、络、俞、募穴相关神经元的形态学和化学特征。通过观察周围和中枢神经系统中传入、传出神经元对肾及其相关特定穴的支配规律, 在细胞水平上揭示运动、感觉和自主神经系统与它们之间的多方位联系。本研究成果将为进一步阐明经穴与脏腑相关性的科学原理提供神经解剖学依据，同时对针灸临床合理选用特定穴治疗相关脏腑疾病具有指导意义。

四年来我们围绕肾及其原、络、俞、募穴相关神经元的形态学和化学特征这条研究主线开展了广泛和深入的研究。首先，从形态学入手综合运用新一代高敏感神经示踪结合共聚焦显微镜技术分别对肾经原穴（太溪）和络穴（大钟）、其背俞穴（肾俞）和募穴（京门）、以及肾脏和肾上腺的神经支配进行了全面的追踪和观察，明确了与其相关运动和感觉神经元以及交感神经节前和节后神经元节段性和区域性性分布特征，同时提示肾俞与肾脏和肾上腺之间的神经解剖学联系最为密切。在此基础上，运用AF488/594-CTB荧光双标记神经示踪技术系统的揭示了大鼠“肾俞”穴区与肾脏和肾上腺之间的神经解剖学联系，其中脊神经节存在少量的感觉神经元通过分支投射“肾俞”穴区与肾脏和肾上腺之间建立了直接的感觉神经联系，同时也观察到一部分分别与“肾俞”穴区、肾脏和肾上腺相关的交感节后神经元位于同节段的交感链中。这些结果提示我们“肾俞”与肾脏和肾上腺在感觉和交感神经支配方面存在的节段性联系可能是针刺肾俞穴可以调节肾脏和肾上腺功能的神经解剖学途径。为此，我们进一步观察了“肾俞”穴区和肾脏神经支配的化学特征，降钙素基因相关肽（CGRP）标记的感觉神经纤维和络氨酸羟化酶（TH）标记的交感神经纤维广泛存在于“肾俞”穴区和肾脏包膜，从另一个侧面提示了它们之间神经解剖学联系的化学特征。总之，我们综合运用新一代高敏感神经示踪和免疫荧光组织化学技术并结合共聚焦显微镜技术深入研究了肾及其原、络、俞、募穴相关神经元的形态学和化学特征。通过观察周围和中枢神经系统传入、传出神经元对肾及其相关特定穴的支配规律, 在细胞水平上揭示了感觉和交感神经系统与它们之间的多方位联系，这些成果为进一步阐明经穴与脏腑相关性的科学原理提供了神经解剖学依据，对针灸临床合理选用特定穴治疗相应脏腑疾病具有重要的指导意义。