腧穴和脑之间多级神经环路的可视化研究

The correlation of the acupoint and nervous system is one of the key points in the acupuncture science. It is not only an important content of local anatomy of acupoints in the text of acupuncture, but also includes various aspects of clinical and basic studies on acupuncture. The former still stayed at the level of gross anatomy, and the latter paid more attention on the effect of acupuncture on one part of the nervous system, however, it is absent of the systemic study on the origination and neural circuits of the innervation of acupoints. Together the previous results of clinical and basic studies at home and abroad, we considered that the innervation of acupoints plays an important role in acupuncture action through sensory, motor, and autonomic neural circuits between the peripheral nervous system and central nervous system. Based on this consideration, the projection and distribution of the nerve fibers and neurons associated with the different acupoints are orderly traced from their locations to the brain in the whole nervous system for determining the specific and common neural circuits of the different acupoints. In this study, the acupoints on the head, trunk, and limbs in the rats were selected as the representative points to label, reveal, and observe the innervations of different acupoints with the conventional neural tracing technique, new generation transsynaptic virus, fluorcent immunohistochemical transparent technique, and combine with laser confocal microscopic technique, for demonstrating and comparing the differences of the correlation of the sensory, motor, and autonomic systems with the different acupoints on the cellular level from the perspective of multi-order neural circuits. Thus, we believe that this project may be not only benefit to understand the neuro-anatomical characteristics of different acupoints and their specific and common active approaches through neural circuits, and also have an important guiding significance to precisely select acupoints and target-treatment as the clinical requirements according to the different diseases.

目前我们对腧穴与神经系统关系的认识主要基于大体神经解剖学的观察，而对支配腧穴神经纤维的源头及其关联的神经环路还缺少深入系统的研究。结合国内外的研究成果，我们认为腧穴相关多级神经环路是针灸借以产生和发挥生物学效应的重要载体之一，它们在外周和中枢之间通过感觉、运动和自主神经系统中的各级神经元进行有序的联接。基于这一认识，本课题将从腧穴局部入手向大脑逐级追踪与不同腧穴相关神经纤维和神经元在神经系统中的投射和分布。我们将以大鼠头面、躯干和四肢处的代表性腧穴入手，综合运用传统神经示踪技术、跨突触嗜神经病毒、免疫荧光组织化学技术和共聚焦显微成像技术对它们的神经支配进行标记、显示和观察，从结构上实现不同腧穴相关的多级神经环路的可视化，在细胞水平上构建“腧穴与脑之间的连接图谱”。通过这项研究，我们将从神经解剖学角度明确不同腧穴作用途径的规律性特征，进而指导针灸临床的精准选穴及其靶向治疗。

目前，我们对腧穴与神经系统关系的认识主要基于大体神经解剖学的观察，而对支配腧穴神经纤维的源头及其关联的神经环路还缺少深入系统的研究。很多研究显示腧穴相关多级神经环路是针灸借以产生和发挥生物学效应的重要载体之一，它们在外周和中枢之间通过感觉、运动和自主神经系统中的各级神经元进行有序的联接。本课题从腧穴局部入手向大脑逐级追踪与不同腧穴相关神经纤维和神经元在神经系统中的投射和分布。我们分别选用大鼠头面、躯干和四肢处的代表性腧穴，综合运用传统和新一代神经示踪技术、免疫荧光组织化学技术和共聚焦显微成像技术对它们的神经支配进行标记、显示和观察，在细胞水平上构建“腧穴与脑之间的连接图谱”。主要研究成果有以下七个方面：（1）从三叉神经节、三叉神经脊束核、三叉神经运动核和面神经核明确了头面部不同腧穴对应的感觉和运动神经环路；（2）从脊神经节、脊髓背角和前角、脑干明确了四肢和躯体不同腧穴对应的感觉和运动神经环路；（3）实验大、小鼠穴位图谱的重新绘制及其标准的制定；（4）“肾俞”穴区与肾脏神经支配的相关性研究取得进展；（5）大鼠后肢相对穴的神经解剖学联系和差异；（6）血清素阳性细胞在大鼠不同穴区皮肤组织中分布的观察；（7）进一步拓展了我们的研究内容，从神经血管单元出发探讨了中风模型的病理特点。通过这项研究，我们从神经解剖学角度明确了不同腧穴作用途径的规律性特征。这些成果对针灸临床的精准选穴及其靶向治疗具有重要的指导意义。