發(fā)布時間：2018-05-06 03:10

  本文選題：抑郁癥 + 慢性不可預知性溫和刺激(CUMS)　； 參考：《重慶醫(yī)科大學》2017年碩士論文

【摘要】：研究背景抑郁癥是一種高度流行且致殘的精神疾病,對人們的生活質量和機體功能有重大損害。然而,我們對抑郁表型及其潛在的病理生理學機制之間的關系仍然知之甚少�，F(xiàn)階段對抑郁癥的研究主要集中在腦區(qū),腦與機體的聯(lián)合探索可能會為抑郁癥病因的全面理解提供新的線索。作為人體最大的內臟器官,肝臟在機體生物轉化,合成,代謝等一系列生理和生化反應中發(fā)揮著重要的作用。但目前仍然沒有用蛋白質組學方法對抑郁模型小鼠肝臟中蛋白表達進行評估的相關研究。目的本研究旨在用iTRAQ標記和LC-MS/MS聯(lián)合的蛋白質組學技術鑒定CUMS抑郁模型小鼠肝臟的蛋白表達譜,用特定的篩選條件尋找相關的差異蛋白,初步分析這些差異蛋白及其所涉及通路的病理生理機制,探討抑郁狀態(tài)下肝臟結構功能改變與蛋白表達譜變化的關系,為診治抑郁癥提供新的思路和方向。方法經(jīng)過環(huán)境適應和一周的糖水訓練之后,我們隨機地把40只正常的C57BL/6J雄性小鼠分為抑郁模型組和正常對照組(20:20)。20只模型組小鼠進行4周的慢性不可預知性溫和刺激之后,將兩組進行行為學評估,若模型成立,從每組中隨機選取小鼠肝臟組織(10:10)進行itraq蛋白組學鑒定,對差異蛋白進行生物信息學分析,另每組選取6只小鼠肝臟組織對差異蛋白和通路進行免疫印跡法(Western blot,WB)驗證。結果1.模型組CUMS之后對兩組的體重,糖水偏好,強迫游泳,曠場試驗等行為學數(shù)據(jù)進行統(tǒng)計學分析,有顯著差異,證明造模成功。2.Itraq串聯(lián)質譜鑒定出4000多種蛋白質,用特殊肽段大于等于2,1.2以上差異倍數(shù),P值小于0.05三個條件,篩選出66種最顯著差異的蛋白質,用于進一步的生物信息學分析。3.通過信號網(wǎng)絡分析(IPA),我們發(fā)現(xiàn)這66種差異蛋白與炎癥反應,免疫調節(jié),脂質代謝和NFκB信號通路等病理生理過程相關。4.此外,通過免疫印跡法(Western blot,WB)驗證了與這些通路密切相關的四種蛋白質,血紅蛋白(HPX),觸珠蛋白(HP),細胞色素P4502A4(CYP2A4)和膽汁鹽磺基轉移酶1(SULT2A1),結果與itraq一致。結論我們的研究首次報道了CUMS抑郁模型小鼠中肝臟組織的蛋白表達譜,為以后研究抑郁癥多層面的機制提供了如肝-腦軸等新的見解。
[Abstract]:Background Depression is a highly prevalent and disabling mental disease, which has great damage to people's quality of life and body function. However, little is known about the relationship between depression phenotypes and their underlying pathophysiological mechanisms. At present, the research on depression is mainly focused on the brain area. The combined exploration of brain and body may provide new clues for the comprehensive understanding of the cause of depression. As the largest visceral organ, liver plays an important role in biological transformation, synthesis, metabolism and a series of physiological and biochemical reactions. However, no proteomics method has been used to evaluate the expression of protein in the liver of depressive mice. Objective to identify the liver protein expression profiles of CUMS depressive mice by iTRAQ labeling and LC-MS/MS proteomics, and to search for differential proteins by specific screening conditions. The pathophysiological mechanisms of these differentially expressed proteins and their related pathways were preliminarily analyzed, and the relationship between the changes of liver structure and function and the changes of protein expression profile in depressive state was discussed, which provided a new way of thinking and direction for the diagnosis and treatment of depression. Methods after environmental adaptation and a week of sugar water training, we randomly divided 40 normal C57BL/6J male mice into depression model group and normal control group 20: 20 mice. After 4 weeks of chronic unpredictable mild stimulation, 40 normal male C57BL/6J mice were randomly divided into depression model group and normal control group. The behavior of the two groups was evaluated. If the model was established, the liver tissues of each group were randomly selected for itraq proteomics identification, and the differential proteins were analyzed by bioinformatics. In addition, the liver tissues of 6 mice in each group were examined by Western blotWB for differential protein and pathway. Result 1. After CUMS, the behavioral data such as body weight, sugar water preference, forced swimming, open field test and so on in the model group were statistically analyzed. The results showed that the model was successful. 2. Itraq tandem mass spectrometry identified more than 4000 proteins. Using the three conditions that the special peptide segment is larger than or equal to 2o 1.2 and the difference multiple P value is less than 0.05, 66 proteins with the most significant difference are selected for further bioinformatics analysis. By signal network analysis, we found that these 66 differential proteins were related to inflammatory response, immune regulation, lipid metabolism and NF 魏 B signaling pathway. In addition, four proteins closely related to these pathways, HPX, HPX, cytochrome P4502A4 (CYP2A4) and bile salt sulfotransferase 1 (SULT2A1), were identified by Western blotDNA (Western blot). The results were consistent with those of itraq. Conclusion our study reported for the first time the expression profile of liver protein in CUMS depression model mice, which provided some new insights into the mechanism of depression in the future, such as liver and brain axis.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R749.4;R-332

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本文編號：1850511