本文關鍵詞： 氧化還原 狀態(tài)液氮低溫環(huán)境 內源性熒光分子 乏氧鼠腦 低溫成像技術　出處：《華中科技大學》2012年碩士論文　論文類型：學位論文

【摘要】：腦組織氧化還原代謝反應的正常進行，是維持腦組織結構完整和功能正常的前提。腦組織氧化還原代謝狀態(tài)的正常維持需要持續(xù)有效的氧氣供應，空氣氧分壓的細微變化都可能導致腦組織氧化還原狀態(tài)的改變，即引起腦細胞線粒體中的NADH和Fp兩種內源性輔酶的濃度發(fā)生改變。本文將內源性熒光分子成像技術和組織氧化還原代謝狀態(tài)低溫速凍、維持技術相結合，，利用多通道低溫斷層熒光顯微成像技術獲取了昆明小鼠全腦在乏氧后的氧化還原代謝狀態(tài)的多截面分布信息，這將有助于人們進一步加深認識不同生理病理狀況下，腦組織的氧化還原代謝情況與其內源性輔酶的關系，為腦氧化還原代謝異常的特異性診斷、治療以及治療后效應的評價提供了方法。 本文主要的工作包括： 1、比較了斷首液氮速凍法、直接整鼠液氮速凍法和漏斗式液氮速凍法三種常見固定實驗鼠腦氧化還原代謝狀態(tài)的方法，并根據樣品結構固定完整程度、樣品氧化還原代謝狀態(tài)的均一性，確定了漏斗法作為適宜本研究的樣品制備技術。 2、液氮低溫環(huán)境可以有效維持生物組織樣品的氧化還原代謝狀態(tài)不變，本文依據液氮低溫技術的基本原理，構建液氮低溫環(huán)境，并自行設計和制作了相應的模塊，確保樣品始終浸潤在液氮環(huán)境中以便在樣品銑削以及成像過程中維持組織樣品氧化還原代謝狀態(tài)不變。 3、在液氮低溫環(huán)境下，采用了多通道低溫三維熒光成像系統獲取了小鼠乏氧和正常生理狀態(tài)下NADH和Fp兩種輔酶濃度的分布信息，結果發(fā)現乏氧鼠腦呈現整體高還原性，并且乏氧鼠腦的白質與灰質之間的區(qū)域差異性與正常鼠腦兩種組織間的區(qū)域差異性剛好相反。
[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.
【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R318.52

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