乏氧鼠脑氧化还原状态的低温成像初步研究

发布时间：2018-01-31 17:56

本文关键词： 氧化还原状态液氮低温环境内源性荧光分子乏氧鼠脑低温成像技术　出处：《华中科技大学》2012年硕士论文　论文类型：学位论文

【摘要】：脑组织氧化还原代谢反应的正常进行，是维持脑组织结构完整和功能正常的前提。脑组织氧化还原代谢状态的正常维持需要持续有效的氧气供应，空气氧分压的细微变化都可能导致脑组织氧化还原状态的改变，即引起脑细胞线粒体中的NADH和Fp两种内源性辅酶的浓度发生改变。本文将内源性荧光分子成像技术和组织氧化还原代谢状态低温速冻、维持技术相结合，，利用多通道低温断层荧光显微成像技术获取了昆明小鼠全脑在乏氧后的氧化还原代谢状态的多截面分布信息，这将有助于人们进一步加深认识不同生理病理状况下，脑组织的氧化还原代谢情况与其内源性辅酶的关系，为脑氧化还原代谢异常的特异性诊断、治疗以及治疗后效应的评价提供了方法。本文主要的工作包括： 1、比较了断首液氮速冻法、直接整鼠液氮速冻法和漏斗式液氮速冻法三种常见固定实验鼠脑氧化还原代谢状态的方法，并根据样品结构固定完整程度、样品氧化还原代谢状态的均一性，确定了漏斗法作为适宜本研究的样品制备技术。 2、液氮低温环境可以有效维持生物组织样品的氧化还原代谢状态不变，本文依据液氮低温技术的基本原理，构建液氮低温环境，并自行设计和制作了相应的模块，确保样品始终浸润在液氮环境中以便在样品铣削以及成像过程中维持组织样品氧化还原代谢状态不变。 3、在液氮低温环境下，采用了多通道低温三维荧光成像系统获取了小鼠乏氧和正常生理状态下NADH和Fp两种辅酶浓度的分布信息，结果发现乏氧鼠脑呈现整体高还原性，并且乏氧鼠脑的白质与灰质之间的区域差异性与正常鼠脑两种组织间的区域差异性刚好相反。
[Abstract]:The normal state of brain redox metabolism is the premise to maintain the integrity and function of brain tissue. The normal state of brain redox metabolism requires a continuous and effective oxygen supply. Slight changes in the partial pressure of oxygen in the air may lead to changes in the redox state of brain tissue. That is to say, the concentration of NADH and FP in mitochondria of brain cells is changed. In this paper, the endogenous fluorescence molecular imaging technology and the state of tissue redox metabolism were frozen at low temperature. In combination with maintenance technique, the multi-section distribution information of redox metabolic state in the whole brain of Kunming mice after hypoxia was obtained by using multi-channel low-temperature tomographic fluorescence microscopy. This will help people to further understand the relationship between the brain redox metabolism and its endogenous coenzymes under different physiological and pathological conditions, which is the specific diagnosis of brain redox metabolism abnormality. Treatment and evaluation of post-treatment effects provide a method. The main work of this paper includes: 1. Three common methods of fixing the brain redox metabolism state of the experimental mice were compared, including liquid nitrogen quick freezing method, direct liquid nitrogen freezing method and funnel liquid nitrogen quick freezing method, and according to the intact degree of sample structure. The homogeneity of the redox metabolism state of the samples was determined. The funnel method was used as the suitable sample preparation technique for this study. 2. The low temperature environment of liquid nitrogen can effectively maintain the state of redox metabolism of biological tissue samples. In this paper, the low temperature environment of liquid nitrogen is constructed according to the basic principle of low temperature technology of liquid nitrogen. The corresponding modules are designed and made to ensure that the samples are always infiltrated in liquid nitrogen environment so as to maintain the same state of redox metabolism in the process of sample milling and imaging. 3. In the low temperature environment of liquid nitrogen, the distribution information of NADH and FP in hypoxia and normal physiological state of mice were obtained by using multi-channel low temperature three-dimensional fluorescence imaging system. The results showed that hypoxic brain showed global hyperreductivity and the regional difference between white matter and gray matter of hypoxic brain was opposite to that between normal brain and white matter.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.52

【共引文献】