【作者】 马铁明；

【导师】 李德新；

【作者基本信息】 辽宁中医学院 ， 中医基础理论， 2004， 博士

【摘要】 随着社会的进步，人均寿命越来越高，人口老龄化已经成为现今世界之普遍现象，预防早衰、延缓衰老、延长寿命、提高生命质量，无疑是目前医学急需解决的世界性问题。中医学在延缓衰老的过程中，积累了丰富的临床经验，并形成了丰富的医学理论，中医学强调脾为后天之本、气血生化之源。临床实践表明，一旦进入脾虚状态，则气血生化乏源，各脏腑的功能逐渐衰退，机体的新陈代谢活动降低或失调，从而出现了与衰老相似的一系列病理变化。 临床实践也证明了人体的生长发育更离不开后天滋养、人体衰老的程度也因后天滋养的不同而有所差别的这一重要事实。即是说人体的生长、发育无不与脾有着密切相关，脾虚是导致机体各种机能逐步减退，呈现衰老状态的基础。因此，深入研究脾虚与衰老的关系，进一步明确其作用机制，在延缓衰老的医学实践中无疑有极为重要的现实意义。 现代科学的发展充分地证明了不断探讨宏观现象下的微观机制，是推动并最终导致科学进步的重要原则。因此，开展中医证候的现代机理研究是弘扬和发展中医药的必由之路。 现代医学认为，衰老的主要特征是全身性的生理功能随着年龄增长而下降，而线粒体是维持细胞以至整个机体生理功能的能源基础，研究表明，线粒体在衰老以及细胞凋亡、程序化死亡等具体过程中发挥着重要的作用，因此说线粒体的衰变是衰老过程的发展基础。 有证据表明代谢旺盛的组织或器官，如心血管系统、中枢神经系统等最先发生衰老表现，就是因为这些部位的mtNDA较多，且最易损伤之故。现在，已经在多种疾病和和衰老过程中发现了mtDNA的突变，而且这种mtDNA构成的改变，可以引起代谢功能的衰变，后者既表现为ATP合成降低，导致细胞以至整个机体生理功能的降低；又表现为自由基等氧化剂的积聚，并成为前者衰变以及全身组织结构衰变的主要原因。因此，美国的Linmane等提出了“衰老的线粒体学说”。 脾虚与衰老密切相关，而线粒体与衰老的关系也日趋明确。研究脾虚与线粒体结构以及功能变化之间的相互关系，有助于揭示脾虚与衰老的内在联系，进而从脾虚这一病理学改变的角度揭示“脾为后天之本”的分子生物学基础，为中医药延缓衰老开辟新的途径。摘要 本项研究在己往研究的基础上，主要研究脾虚证老龄大鼠心、脑组织中mtDNA的缺失变化以及呼吸链复合体酶活性的情况，并结合以往的研究成果，探讨脾虚状态下mtDNA缺失、呼吸链复合体酶活性等的变化规律以及补益脾胃方药干预，阐释其延缓衰老的分子生物学机制。本项研究主要内容包括:1、观察了脾气虚、脾阳虚、脾阴虚三证老龄大鼠模型心、脑mtDNA缺失突变的变化以及补益脾胃方药的干预作用。2、观察了脾气虚、脾阳虚、脾阴虚三证老龄大鼠模型心、脑组织中呼吸链复合酶I、IV酶活性的变化以及补益脾胃方药的干预作用。 研究结果表明:1、老龄大鼠心、脑组织中mtDNA中普遍存在着明显的缺失片段，而在青年 大鼠相应组织中mtDNA中未检测到缺失片段，表明mtDNA的缺失随年龄而积 累。2、脾虚状态下老龄大鼠心、脑组织中mtDNA中也存在明显的缺失片段，且 高于老年空白组，各亚型之间的mtDNA的缺失具有一定的规律性，提示mtDNA 缺失可能是脾虚衰老的重要物质基础之一。3、补益脾胃方药能明显降低老龄大鼠心、脑组织中mtDNA的缺失率，揭示 了补益脾胃之法延缓衰老作用的可能分子机理。4、老龄大鼠心、脑组织中线粒体呼吸链酶复合体活性发生明显改变。与青 年大鼠相比，其活性明显降低，提示衰老状态下，酶活性下降、机体的能量 不足可能是各脏器功能衰退的主要原因之一。S、脾气虚证、脾阳虚证、脾阴虚证状态下，老龄大鼠心、脑组织中酶复合 体活性的不同变化，提示脾虚状态下，其呼吸链功能的变化规律不尽相同。6、补益脾胃方药能明显提高脾虚状态下，老龄大鼠心、脑组织中呼吸链酶 复合体的活性，表明补益脾胃方药可以降低自由基对心、脑组织中mtDNA 的氧化损伤，减少有缺陷的呼吸链，增加细胞所需的能量。7、对老龄大鼠的补益治疗缺失辩证分型，不能取得预期的补益效果，提示 一般补养也应当遵循“辨证论治”原则。 目前，衰老机理的研究已经从整体水平、器官水平、细胞水平发展到分子水平，各种衰老假说从不同的角度诊释着衰老的机制，本项研究结果表明补益脾胃之法可以通过调整mtDNA的缺失状态以及呼吸链复合体酶活性的变化来发挥其补脾延缓衰老的作用，结合以往的研究成果表明补益脾胃方药可以整体性地影响到机体的不同层次、不同器官、乃至不同组织，综合性地发挥其补脾延缓衰老的作用。全面系统地从分子水平来阐明脾虚证的科学内涵，将为开发治疗脾虚证的新方药提供坚实的实验依据，更会为延缓衰老、提高生命质量及防治重大疾病等开辟新的途径。更多还原

【Abstract】 In company with advancement of society, per capita lifetime become more and more long, tendency of aging has become universal phenomenon of present world, it is undoubtedly the most urgent medicinal problem to prevent premature senility, postpone decrepitude, and prolong lifetime and increase quality of life. Traditional Chinese Medicine has abundant clinical experience and form theoretical system to postpone decrepitude, it emphasize that spleen is the root of after-birth and the source of qi and blood. Clinical practice also show that once human body is in the state of spleen deficiency, production of qi and blood would be deficiency, function of each internal organs would be weakened, metabolism would be depressed or maladjustment, then a series of pathology change resemble with decrepitude would appear.It has also been proved by clinical experience that growth and development of human body depend on nourishing of after-birth, the degree of decrepitude is also influenced by nourishing of after-birth. So the growth and development of human body has important relationship with spleen, spleen deficiency is the basis of subsidence of all function and appearance of decrepitude to human body. Therefore, researching the relationship between spleen deficiency and decrepitude and investigating its effective mechanism have most important realistic significance in the course of medical practice to postpone decrepitude.Development of realistic science has proved that researching microcosmic mechanism under macroscopical phenomenon is the important principle to impulse and cause advancement of science finally. So contemporary mechanism research on TCM syndrome or symptom complex is the true way to develop TCM.Modern medicine consider that the main character of decrepitude is descend of systemic physiological function along with increase of age, in the same time mitochondria is not only the energy basis of cell activity but also the energybasis of physiological function of all the body, it has important function in the course of decrepitude, apoptosis and sequencing death, so decline of mitochondria is the foundation of the course of decrepitude.It has been proved that tissue and organ whose metabolism is bloom such as cardiovascular system and central nervous system etc. are the place to appear decrepitude firstly, it because that mtDNA in these place is more and easy to be damaged. Nowadays, in many disease and the course of decrepitude, mutation of mtDNA has been found, and this change of mtDNA formation can cause decline of metabolism function, the latter not only appears reduce of ATP compound which cause physiological function decline of cell and all the body but also appears gathering of oxidizer such as free radical which cause the decline of the former and tissue in all the body. Thereby, Linmane (American) advance "Decrepit Mitochondria Theory" .The relationship of spleen deficiency and decrepitude is osculating, and the relationship of mitochondria and decrepitude is more and more definitude too. The research of relationship between spleen deficiency and structure and function of mitochondria is helpful to reveal the internal contact between spleen deficiency and decrepitude, then reveal molecular biology foundation of "spleen is the basis of after-birth" through pathology change of spleen deficiency, open up new method to postpone decrepitude by Chinese herbs.Based on previous research, this experiment is mainly research the deficiency of mtDNA in heart and brain of old rats with spleen deficiency and the activity of respiratory chain complex kinase. Combine with previous research result, discuss the change disciplinarian of mtDNA deficiency and respiratory chain complex kinase activity, and the molecular biology mechanism of postpone decrepitude by Chinese herbs for reinforcing spleen. The main content of this research includes:1. Observe the change of mtDNA deficiency in heart and brain of old rats with spleen qi deficiency, spleen yang deficiency and spleen yin deficiency, and the intervening fun更多还原