注射阿尔新蓝对透明鱼体内类经脉结构的初步研究

发布时间：2018-09-19 09:33

【摘要】：经络理论是中医基础理论的核心内容之一,两千多年来一直有效地指导着中医临床实践。而对于经络实质的探索在上世纪五六十年代才刚刚起步,国内外学者利用各种现代科学技术从生物物理、解剖、组织、生化等多领域和角度围绕经络开展了大量研究工作,以期揭示经络实质,实现经络的客观显示。以张维波教授等为代表的体液派研究发现,经络的主要结构是有利于组织液流动的间质组织。基于生物流体力学原理,张教授通过一系列实验验证了经络是一种具有低流阻性质的组织液通道。从经络运行组织液的特性出发,通过向组织液通道内注射具有示踪功能的液体可以尝试对经络进行显示,并可以研究其所在的解剖学位置。古典经络理论认为经络有“行血气,营阴阳,决死生,处百病”的作用,也就是说人体和动物的生命活动都是在经络系统的控制和调节下进行的。而且大量针灸学和生物物理学研究也已证实经络系统在人体和动物体上普遍存在。长期以来,在针灸经络的实验研究中所使用的模式生物大多集中于大鼠、小鼠、家兔等哺乳动物,对于与人类亲缘关系较远的鱼类在理论上也应有经络或类似结构的存在,但鱼类在经络研究中则较少使用。我们观察发现,某些种系的鱼,如彩裙鱼、玻璃燕鱼身体半透明,呈侧扁型,具有良好的透光性,对注入其体内的有色示踪染料有良好的显示效果,以利于观察。因此,本研究尝试使用彩裙鱼和玻璃燕鱼作为实验动物,通过向其体内注射阿尔新蓝染料进行示踪的方法,围绕透明鱼体内是否存在类经脉结构进行初步研究。实验一:彩裙鱼体内注射阿尔新蓝染液的侧面和横切面观察目的:通过向彩裙鱼体内注射阿尔新蓝染液,观察染液在彩裙鱼体内迁移后的分布情况,进而探索透明鱼体内是否存在类经脉结构;方法:分别将45条彩裙鱼用三卡因溶液麻醉后,使用微量注射泵在脊柱中点或背鳍下缘缓慢注射25μl浓度为0.1%的阿尔新蓝染液,待染液充分迁移后麻醉处死彩裙鱼,用数码相机和体视显微镜在鱼体侧面拍照观察染液的分布;用包埋剂包埋鱼体,冻结后沿鱼体横切面(即垂直于头-尾轴)将鱼体切成厚度为1mm的切片,拍照观察染液在鱼体横切面的分布。结果:通过观察总结彩裙鱼侧面整体照片发现,染液迁移后出现七条明显的纵行的蓝色线状结构,根据其分布位置分别将其命名为背中线、腹中线、背侧线、侧中线和三条腹侧线。此外,还出现部分较短的横向分支结构。对其切片后观察横切面,发现线状结构大多位于背中隔、腹中隔、肌隔、水平隔膜等疏松结缔组织处,与古典经络理论中人体经脉的循行位置“分肉之间”相似。结论:彩裙鱼体内存在类经脉结构,透明鱼的使用可能为经络的示踪研究提供更方便的动物模型。实验二:彩裙鱼体内注射阿尔新蓝染液的动态迁移观察目的:通过向彩裙鱼体内注射阿尔新蓝染液,观察总结染液在彩裙鱼体内的动态迁移规律;方法:分别将36条彩裙鱼进行与实验一中相同的麻醉和注射方法,在注射过程中将数码相机固定于鱼体正上方录像观察染液的注射和动态迁移过程,根据动态录像分别计算比较染液在背部上方和侧面正中迁移的长度和速度;结果:在背部上方染液和侧面正中染液大多沿头尾方向纵向迁移。统计分析发现,染液在背部上方向头部和向尾部方向的迁移在长度分别为11.30±6.63mm和7.23±4.12mm,迁移速度分别为 2.85±1.62mm/s 和 1.80±0.97mm/s,有极显著性差异(P0.01),在出现侧面正中的迁移中,由注射点向尾部的单向迁移占68.7%,由注射点向头部单向迁移占9.4%,由注射点附近向头尾的双向迁移占21.9%;结论:彩裙鱼背部上方和侧面正中可能存在固有的定向组织液流动,这种定向流动与经脉气血运行具有方向性的特点相似。实验三玻璃燕鱼体内不同流阻点注射阿尔新蓝染液的迁移研究目的:在玻璃燕鱼体内测定低流阻点或高流阻点后注射阿尔新蓝染液,观察比较染液的迁移特点;方法:玻璃燕鱼10条,随机分为低流阻点组和高流阻点组,每组5条分别麻醉后,使用连续流阻仪在玻璃燕鱼体内测定低流阻点或高流阻点,用微量注射泵在低流阻点或高流阻点缓慢注射浓度为0.1%阿尔新蓝染液20μl,待染液充分迁移后麻醉处死玻璃燕鱼,使用数码相机在鱼体侧面拍照观察染液的迁移特点;结果:在低流阻点组玻璃燕鱼体内注射阿尔新蓝染液后,有4条出现了较长距离的线状迁移;在高流阻点组玻璃燕鱼体内注射阿尔新蓝染液后染液多在注射点附近局部扩散,有1条出现了较长距离的线状迁移;结论:染液在低流阻点注射后更容易出现长距离线状迁移的特点初步证明了低流阻通道的流体约束性。
[Abstract]:The theory of meridians and collaterals is one of the core contents of the basic theory of traditional Chinese medicine, which has been effectively guiding the clinical practice of traditional Chinese medicine for more than 2000 years. A great deal of research work has been done on collaterals in order to reveal the essence of the meridians and realize the objective display of the meridians and collaterals. According to the characteristics of tissue fluid in meridians and collaterals, it is possible to show the meridians and collaterals by injecting tracer liquid into the tissue fluid channel, and to study the anatomical location of the channel. That is to say, the life activities of human and animals are controlled and regulated by the meridian system, and a large number of acupuncture and biophysical studies have confirmed that the meridian system is widespread in human and animals. Rabbits and other mammals should also have collaterals or similar structures in theory for fish that are distantly related to humans, but fish are seldom used in the study of meridians and collaterals. Therefore, this study tried to use colored skirt fish and glass swallow as experimental animals, by injecting alxin blue dye into the body of transparent fish to trace the existence of meridian-like structure. Experiment 1: Alxin was injected into the body of colored skirt fish. OBJECTIVE: To observe the distribution of Alcian blue dye solution in colored skirt fish by injecting Alcian blue dye solution into the body of colored skirt fish, and to explore whether there is meridian-like structure in transparent fish. The colored skirt fish were anaesthetized and killed after the dye solution was fully migrated. The distribution of dye solution was observed by digital camera and stereomicroscope on the side of the fish body. Result: Seven distinct blue linear structures were observed and summarized on the whole side of the fish. According to their distribution positions, they were named dorsal midline, ventral midline, dorsal midline, lateral midline and three ventral lines. Observing the transverse section, we found that most of the linear structures were located in the loose connective tissues such as dorsal septum, abdominal septum, muscular septum and horizontal septum, which were similar to the position of meridians in classical meridian theory. OBJECTIVE: To observe the dynamic migration of Alxin blue dye solution by injecting Alxin blue dye solution into the coloured skirt fish, and to observe and summarize the dynamic migration of the dye solution in the coloured skirt fish. The length and speed of dye migration in the middle of the upper back and the side were calculated and compared according to the dynamic video recording. Results: The dye migration in the middle of the upper back and the side was mostly longitudinal along the head and tail direction. Statistical analysis showed that the length of migration of dye solution from upper back to head and tail was 11.30 6.63 mm and 7.23 65507 The unidirectional migration of the shooting point to the head was 9.4%, and the bidirectional migration from the injection point to the head and tail was 21.9%. OBJECTIVE: To observe and compare the migration characteristics of blue dye solution by injecting Alcian blue dye solution after measuring low flow resistance point or high flow resistance point in glassy swallow. At the blocking point or high current blocking point, the glass swallow was anaesthetized and killed with 0.1% Alcian blue dye at the low or high current blocking point by a microinjection pump. The migration characteristics of the dye were observed by photographing the side of the fish with a digital camera. There were four long-distance linear migration after injection of Alcian blue dye in the high flow resistance group, and one of them had long-distance linear migration after injection of Alcian blue dye in the high flow resistance group. The fluid constraint of flow resistance channels.
【学位授予单位】：中国中医科学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R224

【参考文献】