【作者】 金友；

【导师】 刘友章；

【作者基本信息】 广州中医药大学 ， 中医内科学， 2006， 硕士

【摘要】 目的:传统中医长于宏观辨证,侧重于对功能的探讨,微观辨证是其薄弱之处,尤其对于物质组织结构特别是在亚细胞水平上的研究尚欠深入。细胞色素氧化酶是线粒体电子传递呼吸链上的标志酶,对氧有高度亲和力,能够使呼吸链以最大速度发挥作用。为了进一步对脾主运化与线粒体结构和功能状态的关系进行观察和评价,我们在“中医脾-线粒体”相关的理论指导下,运用电镜酶组织细胞化学的方法,探讨消化性溃疡中医证型与线粒体细胞色素氧化酶相关性。 方法:选择消化性溃疡病患者脾气虚证、胃热证及正常组各5例,胃镜下取胃粘膜组织,改良的以3,3-二氨基联苯胺四盐酸（DAB-4HCL）为底物的细胞色素氧化酶细胞化学定位方法孵育组织,再用常规电镜制样,不染色,电镜下观察线粒体结构及细胞色素氧化酶分布情况,并运用体视学方法进行图像分析,探讨不同证型细胞化学反应的特征。 结果:正常组在活性部位可看到细胞色素氧化酶反应颗粒沿线粒体内膜和嵴均匀排列;胃热组线粒体丰富,酶阳性物沉积于多数尚完整的线粒体膜和嵴上,少数线粒体膜或嵴破坏;脾气虚组患者壁细胞不仅线粒体数量减少,而且线粒体的结构有明显的损伤,部分线粒体细胞色素氧化酶染色缺失。与正常组比较,脾气虚组患者胃粘膜壁细胞线粒体面积Am、周长Bm减小（P<0.05）,线粒体发生固缩,线粒体比膜面δ_m值也随之减小（P<0.01）,酶反应面积Ae的值减小（P<0.01）,线粒体细胞色素氧化酶阳性反应密度R_me随之减小（P<0.01）,线粒体膜上酶反应不连续,所以酶阳性反应周长Be的值也减小（P<0.01）;胃热证组患者线粒体面积Am、阳性反应的线粒体膜长度B_e于正常组比较没有显著性意义（P>0.05）,酶反应面积Ae的值减小（P<0.01）,线粒体上酶反应不连续,酶阳性反应密度R_me减小（P<0.01）;脾气虚组Am、Bm、Ae、Be、R_me均低于胃热组（P<0.01）。 结论:消化性溃疡中医不同证型与线粒体细胞色素氧化酶具有相关性。脾气虚组患者线粒体数目减少及细胞色素氧化酶活性降低,导致能量不足,脾主运化失司,引起一系列脾虚证候,这是中医“证”的一个重要超微病理基础。进一步印证了中医脾—线粒体相关理论,从亚细胞水平,较确切的阐明了脾主运化的实质内涵。更多还原

【Abstract】 Objective: Traditional Chinese Medicine excels in macroscopic syndrome differentiation, which lays emphasis on the research of functions while being weak in microcosmic syndrome differentiation, especially lack of profound research on tissue structure of substance on sub-cellular level. Cytochrome oxidase is the marking enzyme on electron transporting respiratory chain of mitochondria, with high affinity on oxygen and can make the respiratory chain react with maximal velocity. In order to make further observation and assessment of the relation of spleen governing transportation, transformation and structure, function of mitochondria, under the guidance of the theory of relevance between spleen of TCM and mitochondria, we chose enzyme histiocytic chemistry method under electron microscope to study the relevance of syndrome of TCM of peptic ulcer and mitochondria cytochrome oxidase.Methods: Separately choosing 5 examples of the patients of peptic ulcer including syndrome of spleen qi deficiency, syndrome of heat in the stomach and normal group. Taking gastric mucous tissue through gastroscope examination, incubating tissues by the methods of cytochemical indication of cytochrome oxidase through improved methods of DAB-4HCL as substrate, making samples by routine electron microscope with non-staining, observing the mitochondria structure and the distribution of cytochrome oxidase through gastroscope examination, making image analysis by stereological methods so as to reflect completely the characteristic of cyto-chemistry reaction of different syndromes.Results: Cytochrome oxidase reactive drop in the active site in normal group lined up evenly along mitochondria inner membrane and crista, There are plenty of mitochondria in the syndrome of heat in the stomach group with cytochrome oxidase reactive drop deposited in the majority of integralmitochondria membrane and crista and a few broken mitochondria membrane and crista. It shows that not only the quantity of mitochondria decreased in the acid cell of patients in syndrome of spleen qi deficiency group, but also with obvious damage in mitochondria structure and abnormal mitochondria and dyeing absence of a little mitochondria cytochrome oxidase (KO. 05), the mitochondria area Am and perimeter Bm of acid cell of gastric mucosa decreased in the patients of syndrome of spleen qi deficiency group compared with the normal group (P(.O. 05), accordingly, mitochondria pyknosis and mitochondria membrane density 8 m value decreased (K0. 01) . Enzyme response area Ae value decreased(FKO. 01) , accordingly, positive reaction density Revalue of mitochondria cytochrome oxidase decreased (P\0. 01) , positive reaction perimeter Be value also decreased because discontinuation of enzyme reaction in mitochondria membrane (K0. 01) ..Mitochondria area Am and positive reactive mitochondria membrane length Be value in patients of syndrome of heat in the stomach group without significance compared with normal group (PX). 05) , enzyme response area Ae value decreased( K0.01 ) , positive reaction density Rm value decreased because discontinuation of enzyme reaction in mitochondria membrane (P\0. 01) , the Am, Bm, Ae, Be and Revalue in spleen qi deficiency group were all lowered compared with heat in the stomach group (K0. 01) .Conclusion: There are correlations between different syndromes of TCM of peptic ulcer and mitochondria cytochrome oxidase. The changes of quality and quantity of mitochondria and cytochrome oxidase in patients of spleen qi deficiency group led to insufficient energy and disfunction of spleen governing transportation and transformation and serial spleen deficiency syndrome. All of these were an important ultra-micro pathological basis of syndrome of TCM. The research confirmed further the theory of relevance of spleen of TCM and mitochondria and elucidated exactly the essence connotation of spleen governing transportation and transformation on sub-cellular level.更多还原