慢性束缚抑郁模型大鼠前额叶代谢组学研究

发布时间：2018-04-10 06:19

本文选题：抑郁症　切入点：慢性束缚　出处：《重庆医科大学》2017年硕士论文

【摘要】：研究背景抑郁症是一种以显著、持久的情绪低落为主要临床特征的常见情感性精神疾病。其发病率、致死率高。长期持续的抑郁症状可导致严重的认知功能障碍,给患者、家庭以及社会带来了沉重的疾病负担。WHO预计,2030年抑郁症将上升为世界第一大负担疾病。尽管大量的科学研究对抑郁症的发病机制进行了探索,提出来了各种机制假说,但是抑郁症确切的发病机制仍然不清。研究表明慢性应激在抑郁症的发病中发挥重要作用,其可导致前额叶(prefrontal cortex,PFC)的结构和功能改变。针对抑郁症患者的脑部影像学研究显示PFC体积减小及活性降低。由此可知,PFC结构功能改变与抑郁症的发生发展高度相关。目的本研究采用非靶向气相色谱-质谱联用技术(gas chromatography-mass spectrometry,GC-MS)分析慢性束缚刺激(chronic restraint stress,CRS)抑郁模型大鼠PFC中代谢物质改变,从而探索抑郁症发病的可能分子机制。方法1.crs抑郁模型构建:领取35只雄性sd(sprague-dawley)大鼠,适应环境一周后,根据糖水偏好试验和(或)运动试验的结果,排除具有异常值的大鼠;剩余的大鼠随机分为crs组和正常对照(non-stressedcontrol,con)组,每天刺激6小时、连续刺激21天后对两组大鼠进行行为学评价;最后,根据糖水偏好试验结果将crs组分为刺激敏感组和刺激不敏感组,仅刺激敏感组和con组用于后期分析。2.采用非靶向gc-ms代谢组学方法检测两组大鼠pfc组织代谢谱改变,采用多变量统计和单变量统计方法,以及benjamini-hochberg校正后,筛选出crs组和con组之间差异代谢物;采用matlab软件对以上差异代谢物进行热图分析。3.采用ipa软件对以上差异代谢物质进行生物信息学功能分析,探索参与抑郁症发病的代谢通路改变。结果1.根据运动试验结果值的95%参考区间和(或)糖水偏好结果值的95%百分位区间,排除5只具有异常值的大鼠;剩余的30只大鼠随机分为crs组(n=20)和con组(n=10);刺激结束后,根据糖水偏好结果,crs组中8只大鼠被归为刺激敏感组、12只大鼠为刺激不敏感组;为保持组间平衡,随机在con组中挑取8只大鼠;最后,16只大鼠(crs敏感组,n=8;con组,n=8)被用于后期分析。2.与con组相比,crs组大鼠的体重增长降低(p0.01)、糖水偏好显著降低(p0.05)、强迫游泳静止时间延长(p0.05),提示CRS组大鼠表现出明显的抑郁样行为;在旷野试验和高架十字迷宫试验中,CRS组和CON组之间无统计学差异,提示CRS组大鼠无焦虑样行为。3.结合多变量统计和单变量统计方法,以及Benjamini-Hochberg校正后,CRS组和CON组大鼠PFC之间发现36种差异代谢物。4.生物信息学功能分析结果显示,这36种差异代谢物主要与氨基酸代谢、能量代谢和脂质代谢有关。结论1.慢性束缚刺激可诱发大鼠抑郁样行为,而无焦虑样行为。2.采用非靶向GC-MS代谢组学方法发现CRS组大鼠的PFC中存在代谢紊乱,这种代谢紊乱主要涉及氨基酸代谢紊乱、能量代谢紊乱和脂质代谢紊乱。此结果为探索抑郁症的发病机制奠定了基础。
[Abstract]:On the background of depression is a significant and persistent low mood as the main clinical features of common thymogenic mental disease. The incidence rate, high death rate. Long term depression can lead to cognitive dysfunction, serious to patients, families and the society has brought a heavy burden of disease.WHO is expected in 2030 will rise for depression the world's largest burden of disease. Although a large number of scientific research on the pathogenesis of depression are explored, proposed various mechanism hypothesis, but the exact pathogenesis of depression is still unclear. Studies show that chronic stress may play an important role in the pathogenesis of depression, which can lead to the prefrontal cortex (prefrontal cortex, PFC) change the structure and the function of patients with depression. According to brain imaging studies showed decreased PFC activity and volume. Thus, the structure of PFC function and depression. The development of students. The purpose of this study is highly correlated with non target gas chromatography-mass spectrometry (gas chromatography-mass spectrometry GC-MS (chronic) analysis of chronic restraint stress restraint stress, CRS) metabolism of PFC in rat model of depression, and to explore the possible molecular mechanism of the pathogenesis of depression. The construction method of 1.crs model of depression receive: 35 male SD rats (Sprague-Dawley), after a week to adapt to the environment, according to the sucrose preference test and (or) exercise test results, excludinghas abnormal values in rats; the remaining rats were randomly divided into CRS group and normal control group (non-stressedcontrol, con), continuous stimulation for 6 hours every day after 21 days of stimulation evaluation performed on two rats; finally, according to the sucrose preference test results of CRS were divided into sensitive and insensitive stimulation stimulation group, only to stimulate the sensitive group and con group for later analysis by.2. Non targeted metabolomics method for detection of GC-MS PFC in the two groups of rats tissue metabolism profiles, using univariate and multivariate statistical statistical methods, and after benjamini-hochberg correction, were selected between CRS group and con group differences in metabolites; the use of MATLAB software for the above differences into the metabolic thermal analysis of.3. on the metabolism of the above differences bioinformatics analysis using IPA software, explore the metabolic pathway is involved in the onset of depression. Results 1. according to the 95% reference interval and exercise test results (or value) 95% percentile interval sucrose preference value results, 5 were excluded with abnormal values in rats; the remaining 30 rats were randomly divided into CRS group (n=20) and con group (n=10); after stimulation, according to the results of the sucrose preference in the CRS group, 8 rats were classified as sensitive stimulation group, 12 rats for stimulation insensitive group; in order to maintain the balance between groups, in the con group were randomly selected 8 rats; finally, 16 rats (CRS sensitive group, n=8; group con, n=8) to be used in the later analysis of.2. compared with con group, CRS group rats reduced weight gain (P0.01), sucrose preference significantly decreased (P0.05), forced to prolong the swimming immobility time (P0.05), suggesting that the CRS rats showed depression like behaviors were in the wilderness; test and elevated plus maze test, there is no significant difference between CRS group and CON group, CRS group of rats.3. combined with univariate and multivariate statistical methods and statistical anxiety like behavior, after Benjamini-Hochberg correction between CRS group and CON group rat PFC found 36 metabolites.4. bioinformatics analysis showed that the 36 main metabolites and amino acid metabolism, lipid metabolism and energy metabolism. Conclusion 1. chronic restraint stress can induce the depressive like behaviors in rats, without anxiety like behavior of.2. with non targeting GC- MS metabonomics revealed that there was metabolic disorder in the PFC of CRS group. This metabolic disorder mainly involved amino acid metabolism disorder, energy metabolism disorder and lipid metabolism disorder. The results laid the foundation for exploring the pathogenesis of depression.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.4;R-332

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