羊腧穴Telocytes形态特征及其与周围结构的关系

【背景】经络是中医理论的基石，而腧穴又是经络线路上的关键节点部位，为中医针灸的实施位点。但是关于腧穴的结构基础与形态组成众说纷纭，未能科学阐明。远细胞(telocytes,TCs)是近年来发现的一种新型间质细胞，最新研究表明其可能是潜在的经络实质细胞，但腧穴处TCs的特征及其分布需要进一步阐明。【目的】分析腧穴和非穴皮肤结构差异，探究腧穴TCs的形态特征，解析TCs与其周围成分的结构联系，为中医针灸治疗的细胞机制研究提供理论支撑。【方法】选择5只成年健康湖羊为试验对象，采集百会(Du₂₀)、曲池(LI₁₁)、三阴交(Sp₆)、膻中(Ren₁₇)、承浆(Ren₂₄)、耳尖(EP₄)等腧穴以及背部和腹部非穴皮肤组织。使用蛋白标记物CD34和Vimentin标记TCs及其突起(telopodes,Tps)、TPS标记肥大细胞、PGP9.5标记神经、TSG101标记胞外囊泡。利用H.E染色和免疫组化技术分析皮肤腧穴和非穴处的结构和微细成分组成，并使用ImageJ和...

Morphological Characteristics of Telocytes at Sheep Acupoints and Its Relationship with Surrounding Structures

【Background】 Meridian theory is the cornerstone of Traditional Chinese Medicine (TCM), and acupoints are the key sites on the meridian, which are the implementation location of the acupuncture. However, there are different opinions on the structural basis and morphological composition of acupoints, which cannot be scientifically clarified. As a newly found type of interstitial cells, Telocytes (TCs) were suggested to be the potential essence cells of the meridian by morphological study recently, but the characteristics and distribution of TCs at acupoints need to be further elucidated. 【Objective】 This study aimed to analyze the structural differences between acupoints and non-acupoints, and to explore the morphological characteristics of TCs at acupoints. The structural relations between TCs and its surrounding components were also analyzed, so as to provide theoretical support for the study of the cellular mechanism of TCM acupuncture treatment. 【Method】 The skin tissues of Baihui (Du₂₀), Quchi (LI₁₁), Sanyinjiao (Sp₆), Danzhong (Ren₁₇), Chengjiang (Ren₂₄), Erjian (EP₄) and non-acupoints on the back and abdomen were collected from five adult healthy Hu sheep. TCs and Tps (telopodes) were stained by specific markers CD34 and Vimentin, the mast cells were labeled by TPS, and nerves were identified by PGP9.5. Extracellular vesicles were marked by TSG101. The structure and fine composition of skin acupoints and non-acupoints were analyzed by H.E and immunohistochemical techniques (IHC), and the morphological quantitative analysis of the data was carried out by using ImageJ and Image-Pro Plus statistical software. The distribution differences of TCs and its related structures at acupoints and non-acupoints were analyzed. On this basis, the morphological characteristics and stereoscopic structure of TCs were observed by transmission electron microscope (TEM), scanning electron microscope (SEM) and immunofluorescence (IF) double labeling technique, and the morphological relationship between TCs and these structures was further analyzed, thus determining the ultramorphology and material basis of acupoints. 【Result】 There were such structures as hair follicles, sebaceous glands, sweat glands and arrector pili muscle, as well as nerves, blood vessels, mast cells, collagen fiber bundles at the acupoints and non-acupoints. However, the number of nerves, blood vessels and mast cells distribution at acupoints was significantly more than that at non-acupoints (P<0.05). More importantly, TCs with slender tubular processes (telopodes, Tps) were distributed in the skins, and the distribution of TCs at the acupoints was significantly different from that of non-acupoints (P<0.05). TCs could be used as the integrator of acupoints stroma. There were extensive relationships between TCs themselves or TCs and surrounding morphological structures (including gap junctions and extracellular vesicles, etc.), which could develop a structural network system. At the ultramicro level, it was observed that the Tps was a typical beaded appearance, which was composed of alternating arrangement of the inflated part (podom, Pd) and the slender stenotic part (podomer, P). The well-developed mitochondria in the cytoplasm of podom, the cellular connection between Tps and Tps, and a large number of extracellular vesicles on or around TCs (significantly more than non-acupoints (P<0.05)) ensured the core role of TCs in the structure of acupoints. Moreover, the structural connection between TCs and epidermal derivatives also verified the relationship between acupoints and epidermal derivatives in the classic book Huangdi Neijing (The Yellow Emperor’s Canon of Internal Medicine) at the cellular level. 【Conclusion】 Structural compositions at the acupoints and non-acupoints were basically the same, while the number of TCs and Tps, nerves, blood vessels, mast cells, extracellular vesicles at acupoints was significantly more than that at non-acupoints; TCs and Tps had the functional structures of connecting and integrating various morphological components, which might be mediators or integrators of different systems at the acupoints. The cell connections among TCs, developed mitochondria and extracellular vesicles had the structural basis for cell communication and energy generation, which corresponds to the “Qi-Xue” in TCM.