DNMT1及血清同型半胱氨酸、叶酸表达变化在卒中后抑郁模型大鼠中的作用

发布时间：2018-06-18 19:59

本文选题：卒中后抑郁 + 大鼠　；参考：《右江民族医学院》2017年硕士论文

【摘要】：目的:建立卒中后抑郁(Post-stroke depression,PSD)大鼠模型,探讨PSD大鼠海马中DNA甲基转移酶1(DN A methyltransferase 1,DNMT1)蛋白、DNMT1mRNA表达与血清同型半胱氨酸(Homocysteine,Hcy)、叶酸(Folic acid,FA)水平的关系及其在PSD发病中的作用。方法:1、将48只雄性Sprague-Dawley(SD)大鼠随机分成正常组、抑郁组、卒中组、PSD组。卒中组应用线栓法制成大脑中动脉闭塞(Middle cerebral artery occlusion,MCAO)模型;抑郁组应用慢性不可测的温和刺激(Chronic unpredictable mild stress,CUMS)结合孤养法制成抑郁模型;PSD组是在MCAO模型的基础上,结合CUMS及孤养法制成PSD模型。2、应用蔗糖水消耗(Sucrose preference test,SPT)试验、旷场试验(Open field test,OPT)以及体质量来评价造模是否成功。3、应用Western blot、qPCR及ELISA法检测各组大鼠海马组织DNMT1蛋白、DNMT1mRNA和血清Hcy、FA的表达水平。结果:1、行为学与体质量检测结果:(1)SPT检测:PSD组糖水消耗量均较其他组减少(P0.01);卒中组与抑郁组糖水消耗量较正常组减少(P0.01);卒中组糖水消耗量较抑郁组减少(P0.01)。(2)OPT检测:1)平面与直立运动:PSD组均较其他组减少(P0.01);卒中组与抑郁组较正常组减少(P0.01);卒中组较抑郁组减少(P0.01);2)中央格停留时间:PSD组均较其他组增多(P0.01);卒中组与抑郁组均较正常组增多(P0.01);卒中组较抑郁组增多(P0.01)。(3)体质量:PSD组均较其他组减低(P0.01);卒中组与抑郁组较正常组减低(P0.01);卒中组较抑郁组减低(P0.01)。2、生化及分生检测结果:(1)Western blot:PSD组海马DNMT1蛋白相对表达量均较其他组低(P0.01);卒中组与抑郁组海马DNMT1蛋白相对表达量均较正常组低(P0.01)。卒中组海马DNMT1蛋白相对表达量与抑郁组比较,差异无统计学意义(P0.05)。(2)qPCR:PSD组海马DNMT1mRNA相对表达量均较其他组低(P0.01);卒中组与抑郁组海马DNMT1mRNA相对表达量均较正常组低(P0.01);卒中组海马DNMT1mRNA相对表达量较抑郁组低(P0.01)。(3)ELISA:PSD组与其他各组比较,血清Hcy水平明显升高,血清FA水平明显降低,差异有统计学意义(P0.01);卒中组、抑郁组分别与正常组比较,血清Hcy水平均明显升高,血清FA水平明显降低,差异有统计学意义(P0.01);卒中组血清FA水平低于抑郁组,差异有统计学意义(P0.01);抑郁组与卒中组的血清Hcy水平比较,差异无统计学意义(P0.05)。3、相关分析结果:Pearson相关分析显示各组大鼠海马DNMT1蛋白、DNMT1mRNA相对表达量与血清Hcy水平均呈明显负相关(P0.01),而与血清FA水平均呈明显正相关(P0.01)。结论:1、MCAO、CUMS及孤养法联合造模的方法能成功制成PSD大鼠模型。2、PSD大鼠海马DNMT1蛋白、DNMT1mRNA的低表达与其血清Hcy水平的升高及FA水平的降低相关,这些表达变化有可能参与了PSD的发病机制。
[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.
【学位授予单位】：右江民族医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3;R749.4;R-332

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