“肺应秋”适应性调控机制的理论和实验研究

【作者】 徐巍华；

【导师】 辛瑛； 马淑然；

【作者基本信息】 北京中医药大学 ， 中医基础理论， 2008， 硕士

【摘要】 中医学对人体调控机理的认识有其独特的方法和手段,如“阴阳对立统一理论、五行生化克制理论和经络调节理论”等,其中蕴含了丰富的调控论思维。古人正是运用这些辩证的思维来认识和治疗疾病,决定了中医对调控机理的认识具有的整体性、宏观性和模糊性等特点,这些体现在中医理论的各个方面。随着现代医学神经神经-内分泌-免疫网络的发现,人们逐渐意识到,人体内部是由众多器官、组织、分子等组成的结构庞大的有机系统而非简单的叠加,各部分通过相互传递信息实现对生理功能的调控,以维持机体的稳态。这些认识同中医学对人体的认识有异曲同工之妙,两者之间存在着很多交叉联系之处,如对中医对五脏功能系统的认识与西医学呼吸、运动、泌尿生殖、消化、循环系统之间存在者很多相似之处,中医对卫气的认识与现代医学的免疫功能类似等等。这些为从中西医结合角度研究脏腑调控理论提供了很好的纽带和桥梁。“肺应秋”是中医藏象学说的重要组成部分,在中医理论体系中居重要的地位。“肺应秋”适应性调控指的是肺脏能够适应秋季的环境变化而调整自身以及与其它脏腑的关系,借以维持机体内环境的稳态,其调控方式与神经-内分泌-免疫密切相关。因此,本课题从机体和肺的免疫防御功能为切入点,从松果腺—褪黑素与肺脏的关系中研究肺与秋季相通应的内分泌免疫学物质基础,并从宏观和微观角度探讨机体免疫功能的变化规律,为中医对脏腑调控理论提供实验依据。实验发现机体和肺的免疫功能存在着季节性变化规律,大部分免疫指标表现为春夏高而秋冬低,这为临床上常见的呼吸系统疾病在秋冬季节高发提供了一定的实验依据,同时在预防和保健医学领域亦具有重要的参考价值。1.目的以“肺应秋”适应性调控机制为研究对象,通过对其进行理论内涵的剖析和实验结果的总结,探索中医藏象本质的思路和方法。主要是通过比较四季褪黑素分泌水平的差异和机体免疫功能的变化,从免疫学角度阐发“肺应秋”的本质内涵。2.方法采用文献加实验研究方法,对中医“肺应秋”调控机制进行理论探讨和实验求证。2.1理论研究采用文献学、逻辑分析等方法,对中医“天人相应”,脏腑适应性调控的理论进行探讨分析,以“肺应秋”为切入点,探讨从免疫学角度进行实验研究的科学性和可行性。2.2实验研究(1)研究思路:本实验研究主要以“肺应秋”相关的神经-内分泌-免疫轴做为切入点,以松果腺摘除动物模型为研究对象,选择了肺局部免疫和体液免疫相关分子做为检测对象,同时观察大鼠在生理状态下和松果腺摘除后各项免疫指标的变化情况,观察松果腺在此过程中的高位调节作用。(2)实验动物:雄性Wistar大鼠,180-200g。随机分为生理组,伪手术组,手术组。自然光照,自由摄取饮食,室温饲养。大鼠由北京维通利华实验动物有限公司提供,于二分、二至前40天购入,饲养一周后行松果腺摘除手术(手术组),伪手术组只结扎矢静脉但不摘除松果腺,术后饲养至二分、二至日取材。(3)检测方法:①肺泡灌洗液中免疫球蛋白sIgA,干扰素-γ(INF-γ):酶联免疫法;②血清褪黑素及肺组织褪黑素测定:酶联免疫法。③血清IL-3、IgG:酶联免疫法。(4)分析方法:所有数据均采用SPSS11.5统计软件进行分析,多组间比较用方差分析,变量关系采用直线相关分析。3.结果3.1理论研究(1)“天人相应”理论的提出具有深厚的时代背景和文化内涵。古代医家通过对人体生命活动的长期观察,在古代哲学的阴阳五行学说和“人与天地相参”的观点启发下,认识到人内在生命活动与自然环境、社会环境之间存在着相互依赖,相互制约的复杂关系,人体五脏系统通过对自身功能活动的调节,从而适应不断变化的外在环境。中医学汲取了周易、道家、法家等诸子百家有关天人合一的思想,结合当时医学发展的需要,在中医哲学层面上提出了“天人相应”的理论。(2)“肺应秋”来源于《内经》中的“时藏阴阳”理论,经过历代的不断充实和发展,逐渐成为中医藏象学说的主要内容。《内经》认为,人类生存于自然界之中,其内在脏腑的生理功能不可避免地受到外界自然阴阳消长的影响而产生相应的改变。基于此,后代医家经过不断总结与发挥,提出了“肺应秋”理论。是指肺的生理功能随着秋季气候的变化而呈现出相应的变化规律,肺的主气、司呼吸、通调水道、主皮毛等功能在秋季表现出与气候变化相适应的适应性调节,从而维持机体内环境的稳态,达到阴阳平衡。(3)肺脏的调控功能主要是从整体宏观角度调节机体的功能状况,免疫系统亦是从机体整体调节入手,研究具体微观物质的变化,二者存在很大的相似性。因此从免疫角度探讨“肺应秋”具有一定的科学基础。另外,大量研究证实,在藏与时的相应及藏对时令的适应性调控机理中,松果腺随季节变化而分泌的褪黑素是中医学藏与时相应的主要调节者。一方面它对自然环境信号因子—光照有灵敏的反应,在光照条件下,松果腺褪黑激素分泌降低,在黑暗条件下,分泌增高。同时它又是重要的神经内分泌换能器,作为一个神经内分泌高位调节器官,对免疫系统有重要的调节作用。因此,松果腺做为我们研究时藏相应的物质中介无疑是一个最佳的选择。3.2实验研究3.2.1季节变化对血清及肺组织MT的影响①春季,生理组与伪手术组比较有显著性差异P<0.05,夏季伪手术组与生理组、手术组有显著性差异P<0.05。血清褪黑素,无论生理组、伪手术组、手术组均可见春分>冬至>秋分>夏至的变化趋势,且春分>秋分,冬至>夏至,摘除松果腺后各季节褪黑素含量下降。②肺组织褪黑素,无论生理组、伪手术组均可见春分>冬至>秋分>夏至的变化趋势,且春分>秋分,冬至>夏至。摘除松果腺后除夏季,其他三季褪黑素含量下降。3.2.2季节变化对全身免疫及肺局部免疫的影响①IFN-γ含量秋分与春分,冬至与夏至之间均有显著性差异P<0.05。摘除松果腺后各季节IFN-γ含量显著降低,与生理组相比具有显著性差异P<0.05。随着季节的变化其在肺泡中的含量呈现出下降趋势,即春>夏>秋>冬。②春分大鼠肺泡灌洗液中sIgA伪手术组与生理组、手术组比较有显著性差异P<0.05,秋分手术组与生理组和伪手术组比较差异极其显著P<0.01。整个呈现出春夏高而秋冬低的趋势。③四季大鼠胸腺指数、脾脏指数均表现为春夏高而秋冬低,两者变化情况是一致的。④生理组大鼠IL-3水平在冬季呈明显的上升趋势,差异极其显著P<0.01,说明在冬季机体的造血功能是增强的,这也与中医“肾应冬”的理论相一致。松果腺摘除后,IL-3含量显著下降,说明褪黑素对于IL-3的增殖具有重要影响。⑤大鼠IgG的含量变化表现为春秋季大致相平,夏季增高而冬季降低,切除松果腺后,IgG呈反应性升高,与生理组比较有显著性差异P<0.05。推测松果腺分泌的褪黑素可能对IgG起抑制性作用。4.结论4.1中医“肺应秋”具有免疫学物质基础本课题从免疫学角度入手,通过动物实验发现机体及肺的免疫功能存在一个季节性变化规律,大部分免疫指标均表现为春夏增高而秋冬降低。肺脏主动适应环境气候变化而产生相应的内部适应机制,肺的宣发功能在秋季下降而肃降作用增强,是导致机体免疫功能降低的主要因素。故认为免疫功能的改变与“肺应秋”存在着密切的关系。4.2“肺应秋”与松果腺的高位调节密切相关本实验研究表明,正常组大鼠血清和肺组织褪黑素的含量随着日照时间的变化而呈现出相应的变化规律,具有明显的光照依赖性特征。机体的免疫功能变化除了呈现出春夏高秋冬低以外,还发现与褪黑素分泌存在偶联关系。即胸腺指数、脾脏指数、血清IgG及肺泡灌洗液sIgA、IFN—γ等均表现为与褪黑素分泌相同或相反的变化趋势,切除松果腺后,这种变化趋势则不明显,组间无显著差异。这可以从某种程度上说明松果腺做为机体的一种高位调节器官,通过其分泌的节律性变化而对机体免疫状态产生影响。4.3“肺应秋”适应性调控机制假说通过本研究和前期的实验成果,我们初步认为“肺应秋”在免疫方面的调控机制如下:季节性光照的变化会形成持续的信号刺激,通过视网膜视神经传递给下丘脑的视交叉上核,然后通过交感神经节后纤维将信息传递给松果腺,从而使松果腺分泌MT产生节律性变化。MT通过血液释放入周身组织器官。通过两种途径激活免疫系统;(1)作用于内源性阿片肽系统而间接激活免疫系统,导致免疫增强。(2)直接作用于免疫器官(如胸腺、脾脏)和免疫细胞(如单核细胞中IL-2和IL-6的表达)发挥其免疫调节作用。在秋季,由于光照周期的缩短,导致了松果腺分泌MT减少,这种节律性的变化会对机体免疫细胞产生持续性的影响,从而使机体免疫功能在秋季表现为“收敛‘的现象。更多还原

【Abstract】 Chinese medicine regulation mechanism of the human body has its own understandingof the theoretical system and effective means of regulation, such as "the unityof opposites of yin and yang theory, the theory of five elements theory, theancients used these macro,movement changes thinking to understand and treatdiseases. As soon as nerve - endocrine - immune regulation theory beendiscovered,pepoles realized that they have many similaties, particularly in themedical understanding of the five internal organs function and West respiratorymedicine, sports, genitourinary, digesting, and the circulatory system .so itprovide us a good link and bridges to research."Lung corroesponds to the Autumn "as the theory of Chinese medicine is animportant component of the theory of Chinese medicine in the Habitat importantposition. "Lung corroesponds to the Autumn "adaptive control theory refers tothe fall of the lungs should be able to adjust their own environmental changeand the relationship with other viscera, in order to maintain the environmentfor the steady-state, its regulation and control methods and neural - endocrine- closely related to immunization. Therefore, this issue from the body’s immunedefense and lung function as the breakthrough point, from the pineal gland - therelationship between melatonin and lung research in the lung and the same shouldbe the fall-immune material basis and from the perspective of macro-andmicro-immune Functional changes in the law.1.Objective"Pulmonary corroesponds to the Autumn" adaptive control mechanism for theresearch object, meaning its theoretical analysis and experimental results ofthe summation of Chinese Zang-Xiang of the nature of research like thinking andresearch methods. By comparing the four seasons and differences in the levelsof melatonin secretion and immune function changes in a coherent exposition"pulmonary corroesponds to the Autumn" nature of the regulation.2. Methods2.1 Theoretical methodsThe literature and experimental research methods, the Chinese " Pulmonarycorroesponds to the Autumn " control mechanisms of TCM theoretical connotationand modern nerve - endocrine - Immune Network’s overall regulation theory, theessence of Chinese Zang-Xiang as empirical studies suggest the feasibility ofideas and channels, and Experimental study on the topic provide some new modeland reference. 2.2 Experimental methods(1) Experimental ideas: The experimental study " Pulmonary corroesponds to the" related nerve - endocrine - immune axis as a starting point, select the pinealgland removed animal model as study targets, a pulmonary and humoral immunityas a related molecule detection object,the rats were simultaneously observed inthe physical condition after removal of the pineal gland and the immune parameterschange, and observe the pineal gland in the course of high regulation.(2) Animal: Male Wistar rats, 180-200g. Physiological were randomly divided intogroups, pseudo-operation group, operation group. Natural light, free intake diet,keep at room temperature. Rats from Beijing Lihua-dimensional experimentalanimals Limited company, 40 days before the Solar Terms, a week later, keepingthe pineal gland removal surgery (surgery), pseudo-operation group only vectorvein ligation pineal but not removed gland, rats were bred after the operationand were decollated at night on the Spring equinox,Summer Solstice,Autum equinoxand Winter Solstice.(3) Detection Methods:①lavage fluid in sIgA immunoglobulin, interferon-γ(INF-γ): ELISA method;②serum melatonin and lung tissue of melatonin: Theenzyme-linked immunosorbent assay.③serum IL-3:enzyme-linked immunosorbentassay; IgG Determination: immune Turbidimetry.(4) Analysis: All data are used SPSS11.5 statistical software for analysis.Multi-group comparison with analysis of variance,linear correlation betweenvariables.3.Results3.1 Resultus of theoretical study(1) Ancient physicians on human life through the activities of the long-termobservation, in the ancient philosophy of Yin and Yang and the five elementstheory "and the world of the Senate," inspired by the view, the inner lives ofpeople recognize activities and the natural environment, social environmentexists between interrelated, mutual checks complex relationship. Five Viscerasystem of the human body through the activities of the self-regulation function,which adapt to the changing external environment.(2) "Pulmonary corroesponds to the Autumn" comes from "HuangDi NeiJing" in the"organ law" theory, has a profound historical background and culturalconnotations. It holds that, in the nature of human existence, its internal organsof the physiological function inevitably be outside the natural rise and fallof the yin and yang from the impact of the corresponding change. Based on this,"the lung corroesponds to the Autumn" refers to the physiological function ofthe lung with the fall of climate change and show a corresponding variation of the main pulmonary gas, the Secretary breathing, clear water diversion Road, themain features such as fur show in the autumn climate adapt to changes in theadaptability of regulation, in order to preserve the environment for thesteady-state, to balance Yin and Yang.(3)Large number of studies confirmed that in the corresponding dirty with thedirty on the season and the adaptability of the mechanism of control, with theseasons change pineal gland secretion of melatonin and is the medical dirty withthe corresponding principal regulator. On the one hand, its natural signal factor- a light sensitive reaction in the light conditions, the pineal gland secretionof melatonin decreased in the dark conditions, increased secretion. At the sametime it is important neuroendocrine transducer, as a high neuroendocrineregulation organs, the immune system has an important role.3.2 Results of experiments3.21 Seasonal variation of serum and lung tissue of melatonin.①spring, physical group and pseudo-operation group more significantdifference,P<0.05(P=0.04).Summer pseudo-surgery group and physiological groupsand the group has significant difference, P<0.05.Serum melatonin, whetherphysical group, pseudo-operation group, surgery group that spring equinox>WinterSolstice> Autumnal Equinox>summer solstice changes in the trend. And the springequinox> Autumnal Equinox, Winter Solstice> summer solstice. And the pineal glandremoved after each season melatonin content decreased.②melatonin lung tissue, whether physical group, pseudo-surgery group thatspring equinox> Winter Solstice> Autumnal Equinox> summer solstice changes inthe trend. And the spring equinox> Autumnal Equinox, Winter Solstice> summersolstice. And the pineal gland removed in addition to the summer after the otherthree quarters of melatonin content decreased.3.22 Seasonal changes effect the Systemic Immune and pulmonary to systemicimmune.①IFN-γlevels in each group there was no significant difference, but we can seea trend, that is, in addition to surgery group than in spring, summer solsticelow, and the other group with its seasonal variations in the content of alveolarshowing a downward trend, Spring > summer> autumn> winter.②Spring equinox in rat alveolar lavage fluid sIgA pseudo-surgery group andphysiological groups and the group there were significant differences (P <0.05),Autumnal Equinox surgery group and the group and pseudo-physiological differencebetween the surgery group and significantly (P <0.01). Throughout the spring andsummer showing a higher level contrast to a lower level of autumn and wintertrends. ③index seasons rat thymus, spleen showed the index for the spring and summerand autumn and winter high-low, the two changes is the same.④This case found that the rats physiological levels of IL-3 in the winter showedthe upward trend,the difference is significant(P<0.01), indicating that in thewinter the body of the hematopoietic function is enhanced, with Chinese medicine"kidney to Winter "consistent with the theory. Pineal gland removed, IL-3 wassignificantly decreased, the melatonin for the proliferation of IL-3 has animportant influence.⑤Rat IgG changes in the content of the performance of rough-spring and autumn,summer and winter increased lower pineal gland resection, lower IgG response wasthat the secretion of melatonin secretion reduce IgG had an impact.3.23 Pineal gland in rats of various immune indicatorsAfter resection of the pineal gland,lung tissue and serum melatonin secretiondecreased significantly and the rhythm of the disappeared. Immune function inaddition to showing a lower in autumn and winter than in spring and summer, alsofound that black and push-secretion coupling relationship exists. That the thymusindex,spleen index,SigA,serum IgG and so IFN-γsecretion of melatoninperformance consistent with the trend of changes. Pineal gland resection, suchtrends do not change significantly, there was no significant difference betweenthe two groups. This can to some extent that the body of the pineal gland as ahigh regulate organ, through its secretion of rhythm changes the immune statusof impact.4. Conclusions4.1 Chinese " Pulmonary corroesponds to the Autumn " has the basis ofimmunological substances.The topics start from the point of view of immunology, through animalexperiments, we found that the immune function of the body and lung there is atrend of seasonal changes, most of immune performance indicators increased inspring and summer, decline in autumn and winter. Lung actively adapt to theenvironment and climate change to a corresponding internal mechanism to vent thepulmonary function decline in the autumn enhanced role Su drop as a result ofreduced immune function of the main factors. It is fact that the changes in immunefunction has a close relation to the "lung corroesponds to the Autumn".4.2 "Pulmonary corroesponds to the Autumn" has closely relation to thehigh-conditioning of Pineal gland.This study shows that normal rat serum and lung tissue Melatonin in thesunshine with the change of time and showing a corresponding change in the law,with obvious light-dependent characteristics. After resection of the pineal gland, lung tissue and serum melatonin secretion decreased significantly and therhythm of the disappeared. Immune function in addition to showing a low in autumnand winter but high in spring and summer, we also found that black andpush-secretion coupling relationship exists. That the thymus index, spleen index,SigA, serum IgG and so IFN-γsecretion of melatonin performance consistent withthe trend of changes. Pineal gland resection, such trends do not changesignificantly, there was no significant difference between the two groups. Thiscan to some extent that the body of the pineal gland as a high regulate organ,through its secretion of rhythm changes and the immune status of impact.4.3 "Pulmonary corroesponds to the Autumn" adaptive control mechanism.Through this study, and preliminary experimental results, we believe thatthe initial "pulmonary corroesponds to the Autumn" in the immune regulationmechanism is as follows: seasonal light changes will be a continuing stimulussignals through the optic nerve transfer to the retina of the hypothalamussuprachiasmatic nucleus, Then sympathetic postganglionic fiber will be conveyingthis message to the pineal gland, thereby enabling a pineal gland secretion MTrhythm changes. MT is released into the blood through the organ. Throughactivation of the immune system in two ways; (1) the role of endogenous opioidpeptides in the system and, indirectly, activates the immune system, leading toimmune enhancement. (2) a direct role in the immune organs (such as the thymus,spleen) and immune cells (such as monocyte IL-2 and IL-6 expression) play itsrole in immune regulation. In the autumn, because of the shortened cycle of light,resulting in the pineal gland secretion MT decline, and that the rhythm of changesto the body’s immune cells to produce sustained impact, so that the immunefunction in the autumn performance of "convergence ’phenomenon.更多还原