【作者】 李俊；

【导师】 邓伟民；

【作者基本信息】 广州中医药大学 ， 中西医结合临床， 2010， 硕士

【摘要】 目的：脾为后天之本,气血生化之源,在生命活动过程中占有重要地位。脾虚证是中医脾胃病常见的病证之一,若脾虚失运,则脏腑、肌肉无以充养、各器官的功能活动则会减退。长期以来,脾虚证一直是人们研究的热点。本研究旨在对溃疡性结肠炎患者脾胃气虚组、大肠湿热组及正常对照组肠上皮柱状细胞线粒体的体视学进行比较,为中医“脾-线粒体相关”理论提供线粒体变化的体视学依据,从而丰富中医“脾-线粒体相关”理论,完善脾胃学说的研究,揭示中医脾的本质。方法：广州军区广州总医院住院及门诊患者共18例,参照排除标准,选择病例,分脾胃气虚组、大肠湿热组证和正常对照组,分别为6例、7例和5例；在肠镜下取肠粘膜组织,常规电镜制样、半薄切片定位后行超薄切片；按体视学原理和方法在电镜下随机摄片,用Image-Pro图像分析软件设计方网测试系统进行测试。按体视学公式分别计算它们的下列参数：体积密度(Vv)、表面积密度(Sv)、数密度(Nv)、平均体积(V)、平均表面积(S)、平均自由程(λ)、表面积与体积比(Rs,)。结果：通过投射电镜观测到：大肠湿热组线粒体较丰富,少数线粒体膜或嵴破坏；脾虚组不仅线粒体数量减少,而且线粒体的结构有明显损伤,可见线粒体肿胀、膜缺损、嵴断裂、基质变淡等超维结构的改变。线粒体的体视学定量测试结果也显示：与正常组比较,脾虚组细胞线粒体的Nv减小,差值较大,差异亦有显著性(P<0.05),λ增大,差值较大,差异亦有显著性(P<0.05),脾虚组线粒体分布明显稀疏,单位体积胞质内线粒体数目明显减少；脾虚组Rsv减小,差值较大,差异亦有显著性(P<0.05),脾虚组线粒体结构发生肿胀等形态变化；S和V值增大(P<0.05)；与大肠湿热组比较Nv减小(P<0.05)、λ增大(P<0.05)、Rsv减小(P<0.05)、V增大(P<0.05)、Sv减小(P<0.05)、V,减小(P<0.05)。结论：本课题运用图像分析技术和现代体视学技术,从三维水平上较系统地定量的对溃疡性结肠炎患者脾虚证线粒体形态结构的基本特点和变化规律进行揭示,结果显示,无论是从体视学密度参数数密度(N,)、体积密度(Vv)、表面积密度(Sv),形状参数比表面和平均表面积(S),尺寸参数平均体积,分布参数平均自由程(λ)等,都表明脾虚组线粒体数目减少,分布明显稀疏,结构发生肿胀、变形等。本课题所采用的现代体视学技术,避免了既往对于细胞形态结构采用定性或半定量的描述方法所带来的人为的误差和不确定性,使结果更具客观性和准确性,结果也进一步印证了中医“脾一线粒体相关”理论,从亚细胞水平,较确切的阐明了脾本质的实质内涵。更多还原

【Abstract】 Speciality:Clinical Digesty of TCMAuthor:Li junTutor:Deng Wei-MinObjective:The spleen lopment, and occupies an important position in the course of life activity. Spleen qi-deficiency is one of common disease medicine evidence in spleen and stomach disease, if the spleen loss operation, the viscera, muscle to fully support, the functional activities of organs will be diminished. For a long time, spleen-deficiency syndrome has been a hot subject people have been studying. This study is compare Columnar intestinal epithelial cells mitochondrial stereological comparisonspleen of the spleen qi-deficiency group, the damp-heat in large bowel group and normal contrastive group in patients with ulcerative colitis, Which will provide the based provides the materiel basis of the acquired constitution, is the source of growth and deveon stereological to enrich Traditional Chinese Medicine "Pi-mitochondria-related" theory, to perfect stomach spleen theory and to reveal the nature of Traditional Chinese Medicine Spleen.Method:A total of 18 cases of hospitalization and outpatient in Guangzhou General Hospital Of Guangzhou Miltjtary Command, refer to having bordered the exclusive standard to choose cases which are divided into the spleen qi-deficiency syndrome group, the damp-heat in large bowel syndrome group and the normal contrastive group.6 cases for spleen qi-deficiency syndrome group, 7 cases for damp-heat in large bowel syndrome group,5 cases for normal contrastive group. Mucosal tissue was obtained from the endoscopy. The views were photographed stochastically in the electron microscopy according to the stereological method and the test was done using the testing system of the square gridding made by the Image-Pro analytical software. With the plasm as the reference space of the mitochondrion, their parameters of volume density (Vv), surface density (Sv), numerical density (Nv), mean volume(V), mean surface(S), mean free distance (λ)and ratio of surface to volume (Rsv)were acquired according to the stereologic formulas. Their differences among different groups were compared.Results:Observed by transmission electron microscopy:the number of mitochondria of the damp-heat in large bowel group rich, or a small number of mitochondrial cristae damage; the spleen qi-deficiency group not only reduce the number of mitochondria, and mitochondria structure was injured, mitochondria swelling, membrane defects, ridge fracture, matrix and other ultra-pale dimensional structure changes. Quantitative stereology of mitochondria results also showed that:the Nv of mitochondrial decreased in spleen qi-deficiency group compared with normal group (P<0.05), accordingly,λincreased (P<0.05), in spleen qi-deficiency group obviously sparse distribution of mitochondria, the number of mitochondria per unit volume decreased; spleen qi-deficiency group Rsv decreased (P<0.05), Swelling of mitochondria structure; the number of S and V increased (P<().05);with colorectal damp-heat in large bowel group Nv decreased (P<0.05),λincreased (P<0.05), Rsv decreased (P<0.05), V increased (P<0.05), Sv decrease (P <().05), Vv decreased (P<0.05).Conclusion:The subject from the three-dimensional quantitative reveal a more systematic basic featuresand and variation of mitochondrial morphology of spleen qi-deficiency group and normal group, using stereological and image analysis methods to quantitatively study the three-dimensional structure of mitochondria of spleen qi-deficiency. The results showed that:mitochondrial distribution of spleen qi-deficiency group were obviously sparse, the number of mitochondria per unit volume decreased, spleen group mitochondria were swelling and deformation than normal group and damp-heat in large bowel group. The changes of quality and quantity of mitochondria 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