本文選題：卒中后抑郁 + 大鼠��； 參考：《右江民族醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:建立卒中后抑郁(Post-stroke depression,PSD)大鼠模型,探討PSD大鼠海馬中DNA甲基轉(zhuǎn)移酶1(DN A methyltransferase 1,DNMT1)蛋白、DNMT1mRNA表達(dá)與血清同型半胱氨酸(Homocysteine,Hcy)、葉酸(Folic acid,FA)水平的關(guān)系及其在PSD發(fā)病中的作用。方法:1、將48只雄性Sprague-Dawley(SD)大鼠隨機(jī)分成正常組、抑郁組、卒中組、PSD組。卒中組應(yīng)用線栓法制成大腦中動脈閉塞(Middle cerebral artery occlusion,MCAO)模型;抑郁組應(yīng)用慢性不可測的溫和刺激(Chronic unpredictable mild stress,CUMS)結(jié)合孤養(yǎng)法制成抑郁模型;PSD組是在MCAO模型的基礎(chǔ)上,結(jié)合CUMS及孤養(yǎng)法制成PSD模型。2、應(yīng)用蔗糖水消耗(Sucrose preference test,SPT)試驗、曠場試驗(Open field test,OPT)以及體質(zhì)量來評價造模是否成功。3、應(yīng)用Western blot、qPCR及ELISA法檢測各組大鼠海馬組織DNMT1蛋白、DNMT1mRNA和血清Hcy、FA的表達(dá)水平。結(jié)果:1、行為學(xué)與體質(zhì)量檢測結(jié)果:(1)SPT檢測:PSD組糖水消耗量均較其他組減少(P0.01);卒中組與抑郁組糖水消耗量較正常組減少(P0.01);卒中組糖水消耗量較抑郁組減少(P0.01)。(2)OPT檢測:1)平面與直立運動:PSD組均較其他組減少(P0.01);卒中組與抑郁組較正常組減少(P0.01);卒中組較抑郁組減少(P0.01);2)中央格停留時間:PSD組均較其他組增多(P0.01);卒中組與抑郁組均較正常組增多(P0.01);卒中組較抑郁組增多(P0.01)。(3)體質(zhì)量:PSD組均較其他組減低(P0.01);卒中組與抑郁組較正常組減低(P0.01);卒中組較抑郁組減低(P0.01)。2、生化及分生檢測結(jié)果:(1)Western blot:PSD組海馬DNMT1蛋白相對表達(dá)量均較其他組低(P0.01);卒中組與抑郁組海馬DNMT1蛋白相對表達(dá)量均較正常組低(P0.01)。卒中組海馬DNMT1蛋白相對表達(dá)量與抑郁組比較,差異無統(tǒng)計學(xué)意義(P0.05)。(2)qPCR:PSD組海馬DNMT1mRNA相對表達(dá)量均較其他組低(P0.01);卒中組與抑郁組海馬DNMT1mRNA相對表達(dá)量均較正常組低(P0.01);卒中組海馬DNMT1mRNA相對表達(dá)量較抑郁組低(P0.01)。(3)ELISA:PSD組與其他各組比較,血清Hcy水平明顯升高,血清FA水平明顯降低,差異有統(tǒng)計學(xué)意義(P0.01);卒中組、抑郁組分別與正常組比較,血清Hcy水平均明顯升高,血清FA水平明顯降低,差異有統(tǒng)計學(xué)意義(P0.01);卒中組血清FA水平低于抑郁組,差異有統(tǒng)計學(xué)意義(P0.01);抑郁組與卒中組的血清Hcy水平比較,差異無統(tǒng)計學(xué)意義(P0.05)。3、相關(guān)分析結(jié)果:Pearson相關(guān)分析顯示各組大鼠海馬DNMT1蛋白、DNMT1mRNA相對表達(dá)量與血清Hcy水平均呈明顯負(fù)相關(guān)(P0.01),而與血清FA水平均呈明顯正相關(guān)(P0.01)。結(jié)論:1、MCAO、CUMS及孤養(yǎng)法聯(lián)合造模的方法能成功制成PSD大鼠模型。2、PSD大鼠海馬DNMT1蛋白、DNMT1mRNA的低表達(dá)與其血清Hcy水平的升高及FA水平的降低相關(guān),這些表達(dá)變化有可能參與了PSD的發(fā)病機(jī)制。
[Abstract]:Objective: to establish a rat model of post-stroke depression (PSD) and to investigate the relationship between the expression of DNMT1 mRNA and serum homocysteine, folate Folic acid FAA in hippocampus and its role in the pathogenesis of PSD. Methods: one, 48 male Sprague-Dawley SD rats were randomly divided into normal group, depression group, stroke group and PSD group. The middle cerebral artery occlusion of middle cerebral artery occlusion (MCAO) model was established in stroke group, and the depression group was treated with chronic unmeasured mild stimuli (Chronic unpredictable mild stress CUMS) combined with solitary therapy. The PSD model. 2 was made with CUMS and solitary culture method. The sucrose water consumption preference test was used to test the PSD model. Open field test (OPTT) and body mass were used to evaluate whether the model was successful or not. Western blottqPCR and Elisa were used to detect the expression of DNMT1 protein and serum HcyT-1 in hippocampus of rats in each group. Results: 1, Behavioral and body mass test results: 1 / 1 SPT Test: the consumption of sugar water in the PSD group was lower than that in the other groups; the consumption of sugar water in the stroke group and the depression group was lower than that in the normal group; and the consumption of sugar water in the stroke group was lower than that in the depression group. Compared with the other groups, the face and upright movement PSD group decreased P0.01U; the stroke group and depression group decreased the P0.01D; the stroke group decreased P0.01D; the stroke group decreased P0.01D (2) the central lattice residence time of the PSD group increased compared with the other groups; the stroke group and the depression group increased the P0.01g time compared with the normal group. The patients in the stroke group and the depression group both increased compared with the normal group. The rest of the patients in the stroke group and the depression group were significantly higher than the normal group. Compared with the depression group, the body weight of the stroke group was higher than that of the depression group. The body weight of the stroke group was lower than that of the other groups. The body weight of the stroke group was lower than that of the other groups, and that of the stroke group and the depression group was lower than that of the normal group. The relative expression of DNMT1 protein in the hippocampus of the stroke group was lower than that of the depression group, and the biochemical and meristem tests showed that the DNMT1 protein expression in the hippocampus of the stroke group was lower than that of the depression group, and the expression of DNMT1 protein in the hippocampus of the control group was lower than that of the depression group. The relative expression of DNMT1 protein in hippocampus of stroke group and depression group was lower than that of normal group. Compared with depression group, the relative expression of DNMT1 protein in hippocampus of stroke group was higher than that of depression group. The relative expression of DNMT1 mRNA in hippocampus of PSD group was lower than that of other groups, the relative expression of DNMT1 mRNA in apoplexy group and depression group was lower than that in normal group, the relative expression of DNMT1 mRNA in hippocampus of stroke group was lower than that of depression group, and the relative expression of DNMT1 mRNA in apoplexy group was lower than that in depression group. The levels of serum Hcy and FA in stroke group and depression group were significantly higher than those in normal group, and the serum FA level was significantly lower than that in stroke group and depression group (P < 0.01), but the serum Hcy level was significantly higher and the serum FA level was significantly lower in stroke group and depression group than in normal group. The serum FA level in stroke group was significantly lower than that in depression group (P 0.01), and the serum Hcy level in depression group was higher than that in stroke group. There was no significant difference (P 0.05). The results of correlation analysis showed that the relative expression of DNMT1 protein and DNMT1 mRNA was negatively correlated with serum Hcy level, but positively correlated with serum FA level. Conclusion the low expression of DNMT1 mRNA in hippocampus of PSD rat model can be successfully established by the combination of MCAOCUMS and solitary nourishment. The low expression of DNMT1 mRNA in hippocampus of PSD rats is related to the increase of serum Hcy level and the decrease of FA level. These changes may be involved in the pathogenesis of PSD.
【學(xué)位授予單位】：右江民族醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R743.3;R749.4;R-332

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