He Wei, Wu Meiling, Jing Xiang-hong, Bai Wanzhu, Zhu Bing, Yu Xiaochun
Zhongguo Zhen Jiu. 2015 Nov;35(11):1181-6.
The present study aims to investigate the kinetic histocytochemical changes of acupoints in different condition. The expression of tryptase (+) mast cells, histamine (HA) , serotonin (5-HT) and nociceptive neuropeptides including calcitonin gene-related peptide (CGRP) and substance P (SP) were observed by immunohistochemistry combined with confocal technology. Mast cells were labeled with anti-mast cell tryptase antibody and simultaneously with HA or 5-HT primary antibodies to observe their co-expression. The results showed that: (1) SP and CGRP were expressed more highly on the cutaneous nerve fibers of "Hegu" (LI 4) after acupuncture stimulation than that of the control. Mast cells aggregated in close proximity to the blood vessels in intra-epidermis and dermis, and some of them with degranulation in the lower dermis and subcutaneous tissue of "Hegu" (LI 4). Both mast cells and their granules appeared with HA (+) and 5-HT (+) expression at stimulated LI 4 sites, while a few intact mast cells with a little expression of 5-HT and HA were distributed in areas of non-stimulated Ll 4. (2) The acupoints in different locations such as Baihui (GV 20), Weishu (BL 21), Zhongwan (CV 12) and LI 4 had the same constituent but the contents were different. (3) The histocytochemical responses of acupoints sensitized by the Gastric mucosa injury (GMI) were also investigated. GMI resulted in neurogenic plasma extravasation by Evans Blue (EB) in the skin of the acupoints over the back and abdomen, which mostly occurred in the T9-T11 dermatomere. The EB extravasation dots just like acupoints sensitization appeared after GMI and disappeared gradually during the natural self-recovery of the gastric mucosa. More SP and CGRP positive nerve fibers were distributed in EB dots than in regions beside EB dots and in the control, mostly distributed in the nerve fibers around both the vessels and root of hair follicle. Mast cells also aggregated and degranulated to release algogenic substances of 5-HT and HA around the vessels in areas of the EB dots. Collectively the acupoints displayed the same histocytochemical responses due to either acupuncture stimulation or GMI. This may potentially be the histocytochemical basis in the local acupoints and acupoints displayed kinetic changes in different condition.
本研究旨在探讨不同状态下穴位的动态组织细胞化学变化。采用免疫组织化学结合共聚焦技术，观察类胰蛋白酶（+）肥大细胞、组胺（HA）、5-羟色胺（5-HT）以及包括降钙素基因相关肽（CGRP）和P物质（SP）在内的伤害性神经肽的表达。用抗肥大细胞类胰蛋白酶抗体标记肥大细胞，同时用HA或5-HT一抗标记，以观察它们的共表达情况。结果显示：（1）针刺刺激后，“合谷”（LI 4）穴皮肤神经纤维上SP和CGRP的表达高于对照组。肥大细胞聚集于表皮内和真皮内的血管附近，在“合谷”（LI 4）穴的真皮下部和皮下组织中，部分肥大细胞出现脱颗粒现象。在受刺激的LI 4穴位处，肥大细胞及其颗粒均呈现HA（+）和5-HT（+）表达，而在未受刺激的LI 4区域，分布着少量表达5-HT和HA的完整肥大细胞。（2）不同部位的穴位如百会（GV 20）、胃俞（BL 21）、中脘（CV 12）和LI 4，其组成成分相同，但含量不同。（3）还研究了胃黏膜损伤（GMI）致敏穴位的组织细胞化学反应。GMI导致背部和腹部穴位皮肤通过伊文思蓝（EB）出现神经源性血浆外渗，主要发生在T9 - T11皮节。GMI后出现类似穴位敏感化的EB外渗点，在胃黏膜自然自我修复过程中逐渐消失。EB点处分布的SP和CGRP阳性神经纤维多于EB点旁区域及对照组，主要分布在血管和毛囊根部周围的神经纤维中。在EB点区域的血管周围，肥大细胞也聚集并脱颗粒，释放5-HT和HA等致痛物质。总体而言，无论是针刺刺激还是GMI，穴位均表现出相同的组织细胞化学反应。这可能是穴位局部的组织细胞化学基础，且穴位在不同状态下呈现动态变化。
