【摘要】：目的：本實(shí)驗(yàn)采用慢性不可預(yù)見性溫和應(yīng)激(chronic unpredictablemild stress，CUMS)建立大鼠抑郁模型，觀察尼美舒利對(duì)慢性應(yīng)激大鼠抑郁行為的影響。采用比較蛋白質(zhì)組學(xué)研究策略，運(yùn)用ITRAQ聯(lián)合LC-MS/MS技術(shù)檢測(cè)正常組及各用藥組大鼠海馬組織與模型組相比較所得的差異蛋白，以期從蛋白質(zhì)組學(xué)角度闡述尼美舒利對(duì)抑郁大鼠的作用機(jī)制并希望找到新的作用靶點(diǎn)。 方法：（1）120只成年雄性SD大鼠，隨機(jī)分為模型組、正常組、陽性對(duì)照組和尼美舒利3,6和12mgkg-1組。正常組不進(jìn)行任何刺激，其余各組采用慢性不可預(yù)見性輕度應(yīng)激方法制備大鼠抑郁模型。造模完成后，分別給予尼美舒利3mgkg-1、6mgkg-1和12mgkg-1，陽性對(duì)照組給予氟西汀3mgkg-1，正常組和模型組給予濃度為0.5%等體積的羧甲基纖維素鈉，每天灌胃1次,連續(xù)21d。 （2）通過高架十字迷宮、曠場(chǎng)實(shí)驗(yàn)和強(qiáng)迫游泳試驗(yàn)，檢測(cè)大鼠行為學(xué)改變；通過組織病理切片HE染色，觀察大鼠海馬神經(jīng)細(xì)胞的形態(tài)變化，通過免疫組織化學(xué)染色，觀察大鼠海馬神經(jīng)細(xì)胞SYN、NGF、BDNF、5-HT1A受體蛋白表達(dá)；通過酶聯(lián)免疫吸附法，檢測(cè)大鼠海馬組織中的IL-1β、IL-4、IL-6、IL-10和TNF-α的含量變化。 （3）采用比較蛋白質(zhì)組學(xué)研究策略，ITRAQ聯(lián)合LC-MS/MS篩選大鼠海馬組織表達(dá)有差異的蛋白質(zhì)。 結(jié)果：（1）模型組大鼠進(jìn)入高架十字迷宮進(jìn)入總次數(shù)和開臂次數(shù)及停留時(shí)間百分率均較正常組減少；強(qiáng)迫游泳實(shí)驗(yàn)中，靜止不動(dòng)的時(shí)間相比正常組增加；曠場(chǎng)實(shí)驗(yàn)中，垂直活動(dòng)次數(shù)和水平活動(dòng)次數(shù)相比正常組減少。模型組大鼠海馬組織細(xì)胞的IL-1β、TNF-α和IL-6含量升高，IL-4和IL-10含量降低；海馬組織中神經(jīng)細(xì)胞出現(xiàn)細(xì)胞丟失、核固縮，SYN、NGF、BDNF及5-HT1A受體蛋白表達(dá)降低。 （2）尼美舒利各劑量組能明顯改善大鼠的抑郁行為，降低海馬組織細(xì)胞IL-1β、TNF-α和IL-6含量，升高IL-4、 IL-10含量。減輕慢性應(yīng)激大鼠海馬神經(jīng)的損傷，阻遏海馬神經(jīng)細(xì)胞BDNF、NGF、SYN、5-HT1A表達(dá)下調(diào)。 (3)與正常組比較，模型組篩選出54個(gè)差異蛋白質(zhì)，41個(gè)高表達(dá)，13個(gè)低表達(dá)。從尼美舒利3mgkg-1組篩選到75個(gè)差異蛋白，55個(gè)高表達(dá)，20個(gè)低表達(dá)。從尼美舒利6mgkg-1組篩選到40個(gè)差異蛋白，31個(gè)高表達(dá)，9個(gè)低表達(dá)。從尼美舒利12mgkg-1組篩選到42個(gè)差異蛋白，12個(gè)高表達(dá)，30個(gè)低表達(dá)。從氟西汀組篩選到70個(gè)差異蛋白，57個(gè)高表達(dá)，13個(gè)低表達(dá)。并且發(fā)現(xiàn)Tyrosine-protein kinase receptor（酪氨酸蛋白激酶受體），Sodium-and chloride-dependent GABA transporter3（SLC6A11）這兩個(gè)差異蛋白在正常組，氟西汀組，尼美舒利3mgkg-1、6mgkg-1和12mgkg-1組篩選出的差異蛋白中均有出現(xiàn)，uniport數(shù)據(jù)庫分析其功能主要與樹突細(xì)胞因子的產(chǎn)生，，炎癥應(yīng)答，細(xì)胞增殖，神經(jīng)的可塑性，腦發(fā)育，神經(jīng)遞質(zhì)的運(yùn)輸?shù)裙δ苊芮邢嚓P(guān)。這兩個(gè)蛋白質(zhì)在憂郁癥的發(fā)病、預(yù)后起到什么作用需要進(jìn)一步深入研究。 結(jié)論：尼美舒利各劑量組對(duì)CUMS大鼠的抑郁行為有明顯的改善作用，其作用機(jī)制可能與抑制炎癥因子的釋放,增加神經(jīng)可塑性有關(guān)。比較蛋白質(zhì)組學(xué)結(jié)果表明海馬組織蛋白質(zhì)組發(fā)生的這種復(fù)雜變化對(duì)抑郁癥的發(fā)病及預(yù)后起到十分重要的作用，兩個(gè)共同蛋白質(zhì)存在正常組與給藥組表明炎性因子及神經(jīng)可塑性相關(guān)蛋白可能是潛在的抗抑郁藥物靶點(diǎn)。
[Abstract]:Objective: To observe the effect of nimesulide on the depressive behavior of chronic stress rats by using chronic unpredictability and stress (CUMS) to establish a rat's depression model. Using the comparative proteomics research strategy, the difference protein obtained from the normal group and the model group was detected by using the ITRAQ combined LC-MS/ MS technique. In order to study the mechanism of the effect of nimesulide on the depressed rats from the group of proteomics, we hope to find a new target of action. Methods: (1) 120 adult male SD rats were randomly divided into two groups: model group, normal group, positive control group and nimesulide 3, 6 and 12mg? kg-Group 1. No stimulation was performed in the normal group, and the rest of the groups were treated with a mild stress method of chronic unpredictability to prepare the rat's depression The model was established. After the model was completed, the dose of sodium methylcellulose with the concentration of 0.5% and the like was given in the normal group and the model group with the concentration of 0.5% and the like in the normal group and the model group, respectively. 1d. (2) The behavior change of the rat's behavior was detected by the overhead cross-maze, the field experiment and the forced swimming test. The morphological changes of the nerve cells in the hippocampus of the rat were observed by the HE staining of the tissue. The SYN and NGF of the neurons in the hippocampus of the rat were observed by immunohistochemical staining. The expression of IL-1, IL-4, IL-6, IL-10 and TNF-1 in the hippocampus of rats was detected by enzyme-linked immunosorbent assay. (3) Using the comparative proteomics research strategy, the ITRAQ combined with LC-MS/ MS to screen the rat hippocampal tissue Results: (1) The total number of access to the elevated cross-maze and the number of open arms and the percentage of residence time in the model group were lower than that in the normal group; in the experiment of forced swimming, the time compared with the rest time Increase in normal group; number and level of vertical activity in field experiment The levels of IL-1, TNF-1, and IL-6 in the hippocampus of the model group were increased, and the levels of IL-4 and IL-10 decreased. (2) The levels of IL-1, TNF-1, and IL-6 in the hippocampus were significantly improved. the content of IL-4 and IL-10 is increased, The expression of SYN, 5-HT1A was down-regulated. (3) Compared with the normal group, the model group screened 54 differences Protein, 41 high expression, 13 low expression. 75 differences were selected from the nintedanib 3mg? kg-1 group Isoprotein, 55 high-expression, 20 low-expression. 40 from the nimesulide 6mg? kg-1 group Differential protein, 31 high expression, 9 low expression. 42 differences were selected from the nimesulide 12mg? kg-1 group Isoprotein, 12 high expression, 30 low expression. 70 differences were selected from the fluoxetine group The two different proteins of Tyromine-protein kinase receptor (tyrosine protein kinase receptor), Sodidium-and chloride-dependent GABA trans-3 (SLC6A11) were found in the normal group, the fluoxetine group, the nimesulide 3mg? kg-1, 6mg? kg-1, and the 12mg? kg-1 group. It is mainly related to the production of dendritic cells, the inflammatory response, the cell proliferation, the plasticity of the nerve, the brain, It is closely related to the function of development, the transport of neurotransmitters, etc. The two proteins are in the pathogenesis of depression, Conclusion: The results of this study suggest that the dose group of nimesulide has a significant effect on the depression behavior of CUMS rats, and its mechanism of action may It is related to the inhibition of the release of inflammatory factors and to increase the plasticity of the neuroplasticity. The pathogenesis and prognosis of depression play an important role in the pathogenesis and prognosis of depression.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R965

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