慢性不可预见性应激抑郁模型大鼠海马MKP-1表达及其启动子区组蛋白H3乙酰化状态研究
发布时间：2019-06-04 22:52
【摘要】：背景 抑郁障碍以心境低落为主要临床特征,具有明显慢性化倾向,严重影响患者的身心健康,造成沉重的疾病负担。对抑郁障碍分子生物学机制的了解有助于改进治疗方法,改善预后。近年来,相关研究中,细胞内信号系统逐渐受到人们的重视,越来越多的研究发现,决定神经元增殖、分化、成熟、存活、突起生长等的细胞内信号通路与抑郁障碍密切相关。MAPK信号系统位于胞内,可传导多种细胞外信号至细胞内,引起基因的表达的变化,此系统在中枢神经系统中的活性与抑郁密切相关。有研究提示,作为MAPK系统的负性调节因子之一,脑中MKP-1的过表达可能与抑郁的发生相关。作为表观遗传学修饰机制之一,启动子区组蛋白H3乙酰化可引起基因表达的提高,推测MKP-1启动子区组蛋白H3乙酰化修饰也可能参与抑郁的发生。本研究建立大鼠慢性不可预见性应激抑郁模型,以氟西汀为干预药物,以海马为研究目标脑区,探讨MKP-1表达及其启动子区组蛋白H3乙酰化在抑郁障碍发生中的地位。 目的 1.了解海马区MKP-1表达及其启动子区组蛋白乙酰化状态与抑郁模型大鼠类抑郁样表现的关系。 2.了解氟西汀抗抑郁效应与海马区MKP-1表达及其启动子区组蛋白乙酰化状态的关系。 方法 1.实验动物,分组,及抑郁模型的建立 将行为学评价得分相近的30只SD大鼠随机平均分为3组,对照组(control),刺激组(CUS),刺激+氟西汀组(CUS+fluoxetine)。对照组正常饲养,其余2组给予慢性不可预见性应激共实施42天,制备抑郁模型。 2.药物干预方法 干预药物采用氟西汀,模拟患者给药途径,用灌胃法给药。刺激组给予相同容积的生理盐水灌胃模拟药物干预。 3.行为学评价方法 采用体质量变化、旷场试验、蔗糖水消耗试验对大鼠进行行为学评价。 4.实验室方法 采用免疫组化法了解海马MKP-1蛋白表达状态,采用染色质免疫共沉淀(CHIP)法了解组蛋白乙酰化状态。 5.统计学方法 采用方差分析分析实验数据,用SPSS13.0进行数据处理。采用单因素方差分析(one-way ANOVA)进行比较。以P0.05为差异有统计学意义。 结果 1.行为学评价 实验前,各组大鼠的体质量、水平运动距离、蔗糖水消耗比例差异无统计学意义(P=0.699,P=0.950,P=0.996)。慢性应激21天后,体质量、水平运动距离、蔗糖水消耗比例刺激组(P=0.003,P=0.013,P=0.009)、刺激+氟西汀组(P=0.000,P=0.022,P=0.008)较对照组明显下降。药物干预21天后,刺激组体质量、水平运动距离、蔗糖水消耗比例显著低于对照组(P=0.000,P=0.001,P=0.000)；刺激+氟西汀组水平运动距离、蔗糖水消耗比例较刺激组明显提高(P=0.000,P=0.000)；刺激+氟西汀组和对照组比较差异无统计学意义(P=0.663,P=0.087)；体质量刺激组和刺激+氟西汀组低于对照组,差异有计学意义(P=0.000,P=0.000)。 2.模型大鼠海马MKP-1表达以及氟西汀的作用 刺激组海马CA1、CA3、DG区MKP-1表达较对照组明显提高,差异有统计学意义(P=0.000,P=0.000,P=0.000)。刺激+氟西汀组海马CA1、CA3、DG区MKP-1表达较刺激组明显下降,差异有统计学意义(P=0.002,P=0.009,P=0.000)。各组大鼠海马CA2区MKP-1表达差异无统计学意义(P=0.828)。 3.模型大鼠海马MKP-1启动子区组蛋白H3乙酰化情况及氟西汀的作用 刺激组海马MKP-1启动子区组蛋白H3乙酰化程度较对照组明显提高,差异有统计学意义(P=0.004)。刺激+氟西汀组较刺激组无明显下降,差异无统计学意义(P=0.323)。刺激+氟西汀组高于对照组,差异有统计学意义(P=0.037)。 结论 1.慢性不可预见性应激抑郁模型大鼠海马内MKP-1表达升高与启动子区组蛋白H3乙酰化水平升高,可能参与抑郁症的发病机制。 2.氟西汀能改善抑郁模型大鼠的行为表现并逆转海马内MKP-1过表达,可能是氟西汀抗抑郁机制的靶点之一。
[Abstract]:background Depressive disorder is the main clinical characteristic of depression, which has the obvious chronic tendency, which seriously affects the physical and mental health of the patient, and causes the heavy disease to be negative The understanding of molecular biological mechanisms in the disorder of depression can help to improve the methods of treatment and to improve the pre-treatment In recent years, in recent years, in the relevant research, the signal system in the cell is gradually being paid attention to, and more and more research has found that the signal pathway in the cells, such as neuron proliferation, differentiation, maturation, survival, protrusion growth, and the like, is closely related to the depressive disorder. The MAPK signal system is located in the cell, can conduct various extracellular signals to the cell, cause the change of the expression of the gene, and the activity of the system in the central nervous system is closely related to the depression The results suggest that the overexpression of MKP-1 in the brain may be associated with the occurrence of depression as one of the negative regulation factors of the MAPK system. On the other hand, as one of the epigenetic modification mechanisms, the histone H3 ethylation of the promoter region can cause the increase of gene expression, and it is presumed that the histone H3 ethylation modification of the MKP-1 promoter region may also be involved in the development of depression. To study the relationship between the expression of MKP-1 and the histone H3 of its promoter region in the pathogenesis of depression. Bit. Objective: To study the expression of MKP-1 in hippocampus and the depression pattern of histone in the model of depression. 2. To study the effect of fluoxetine on the expression of MKP-1 in hippocampus and its promoter region histone B systematize A state-to-state relationship. Method 1. Experimental animals Thirty SD rats with similar behavior evaluation scores were randomly divided into 3 groups, control group (control), stimulation group (CUS), stimulation + fluoxetine group (CU). S + fluoxetine. The control group was normally fed and the remaining 2 groups were given chronic unpredictability stress in a total of 42 days, that system Depressive model.2. The drug intervention method intervenes with the use of fluoxetine To mimic the route of administration of the patient and administer it by gavage. Same volume of saline 3. The method of behavior evaluation adopts the change of body mass and the level of the body mass. Field test, sucrose water 4. The expression of the MKP-1 protein in the hippocampus was studied by means of the immunohistochemical method in the laboratory. chromatin immunoprecipitation a statistical method for the understanding of histone ethylation status by means of the (CHIP) method The data of the experiment was analyzed by the analysis of variance, and the data was analyzed by SPSS 13.0. Lemma. One-factor analysis of variance (one- wa y ANOVA) Results 1. The body mass, the horizontal movement distance and the water consumption of sucrose in each group were measured before the behavior evaluation. The proportion difference was not statistically significant (P = 0.699, P = 0.950, P = 0.996). After 21 days of chronic stress, the body mass, the horizontal movement distance, the sucrose water consumption ratio stimulation group (P = 0.003, P = 0.013, P = 0.009), stimulation + Flui The control group (P = 0.000, P = 0.022, P = 0.008) was significantly lower than that in the control group (P = 0.000, P = 0.022, P = 0.008). After 21 days of drug intervention, the mass of the stimulation group, the distance between the horizontal movement and the consumption of sucrose water were significantly lower than that of the control group (P = 0.000, P = 0.001, P = 0.000), and the level of the stimulation + fluorositin group was higher than that in the control group (P = 0.000, P = 0.001, P = 0.000). The percentage of sucrose water consumption was significantly higher than that in the stimulation group (P = 0.000, P = 0.000); the difference between the stimulation and the fluoxetine group and the control group was not statistically significant (P = 0.663, P = 0.087); the body mass stimulation group and the stimulation + Fluoxetine group was lower than that in the control group, and the difference was calculated (P = 0). (000, P = 0.000).2. The expression of MKP-1 in the hippocampus of the model rats and the expression of MKP-1 in the hippocampal CA1, CA3, and DG regions of the stimulated group of fluoxetine The difference was significant (P = 0.000, P = 0.000, P = 0.000). The difference was significant (P = 0.002, P = 0.009, P = 0). 000). There was no significant difference in the expression of MKP-1 in the hippocampal CA2 region of each group (P = 0). (828).3. The model rat hippocampal MKP-1 promoter region histone H3 B and the effect of fluoxetine in the hippocampus MK The level of histone H3 in the P-1 promoter region was significantly higher than that in the control group, and the difference was of statistical significance ( P = 0.004). There was no significant difference in the stimulation group and the stimulation group, and the difference was not statistically significant. to learn from one's The difference was statistically significant (P = 0.323) in the control group (P = 0.323). Conclusion 1. Chronic unpredictability stress The expression of MKP-1 in the hippocampus of the rat hippocampus increased with the level of histone H3 in the promoter region, and may be involved in the pathogenesis of depression.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.4
[Abstract]:background Depressive disorder is the main clinical characteristic of depression, which has the obvious chronic tendency, which seriously affects the physical and mental health of the patient, and causes the heavy disease to be negative The understanding of molecular biological mechanisms in the disorder of depression can help to improve the methods of treatment and to improve the pre-treatment In recent years, in recent years, in the relevant research, the signal system in the cell is gradually being paid attention to, and more and more research has found that the signal pathway in the cells, such as neuron proliferation, differentiation, maturation, survival, protrusion growth, and the like, is closely related to the depressive disorder. The MAPK signal system is located in the cell, can conduct various extracellular signals to the cell, cause the change of the expression of the gene, and the activity of the system in the central nervous system is closely related to the depression The results suggest that the overexpression of MKP-1 in the brain may be associated with the occurrence of depression as one of the negative regulation factors of the MAPK system. On the other hand, as one of the epigenetic modification mechanisms, the histone H3 ethylation of the promoter region can cause the increase of gene expression, and it is presumed that the histone H3 ethylation modification of the MKP-1 promoter region may also be involved in the development of depression. To study the relationship between the expression of MKP-1 and the histone H3 of its promoter region in the pathogenesis of depression. Bit. Objective: To study the expression of MKP-1 in hippocampus and the depression pattern of histone in the model of depression. 2. To study the effect of fluoxetine on the expression of MKP-1 in hippocampus and its promoter region histone B systematize A state-to-state relationship. Method 1. Experimental animals Thirty SD rats with similar behavior evaluation scores were randomly divided into 3 groups, control group (control), stimulation group (CUS), stimulation + fluoxetine group (CU). S + fluoxetine. The control group was normally fed and the remaining 2 groups were given chronic unpredictability stress in a total of 42 days, that system Depressive model.2. The drug intervention method intervenes with the use of fluoxetine To mimic the route of administration of the patient and administer it by gavage. Same volume of saline 3. The method of behavior evaluation adopts the change of body mass and the level of the body mass. Field test, sucrose water 4. The expression of the MKP-1 protein in the hippocampus was studied by means of the immunohistochemical method in the laboratory. chromatin immunoprecipitation a statistical method for the understanding of histone ethylation status by means of the (CHIP) method The data of the experiment was analyzed by the analysis of variance, and the data was analyzed by SPSS 13.0. Lemma. One-factor analysis of variance (one- wa y ANOVA) Results 1. The body mass, the horizontal movement distance and the water consumption of sucrose in each group were measured before the behavior evaluation. The proportion difference was not statistically significant (P = 0.699, P = 0.950, P = 0.996). After 21 days of chronic stress, the body mass, the horizontal movement distance, the sucrose water consumption ratio stimulation group (P = 0.003, P = 0.013, P = 0.009), stimulation + Flui The control group (P = 0.000, P = 0.022, P = 0.008) was significantly lower than that in the control group (P = 0.000, P = 0.022, P = 0.008). After 21 days of drug intervention, the mass of the stimulation group, the distance between the horizontal movement and the consumption of sucrose water were significantly lower than that of the control group (P = 0.000, P = 0.001, P = 0.000), and the level of the stimulation + fluorositin group was higher than that in the control group (P = 0.000, P = 0.001, P = 0.000). The percentage of sucrose water consumption was significantly higher than that in the stimulation group (P = 0.000, P = 0.000); the difference between the stimulation and the fluoxetine group and the control group was not statistically significant (P = 0.663, P = 0.087); the body mass stimulation group and the stimulation + Fluoxetine group was lower than that in the control group, and the difference was calculated (P = 0). (000, P = 0.000).2. The expression of MKP-1 in the hippocampus of the model rats and the expression of MKP-1 in the hippocampal CA1, CA3, and DG regions of the stimulated group of fluoxetine The difference was significant (P = 0.000, P = 0.000, P = 0.000). The difference was significant (P = 0.002, P = 0.009, P = 0). 000). There was no significant difference in the expression of MKP-1 in the hippocampal CA2 region of each group (P = 0). (828).3. The model rat hippocampal MKP-1 promoter region histone H3 B and the effect of fluoxetine in the hippocampus MK The level of histone H3 in the P-1 promoter region was significantly higher than that in the control group, and the difference was of statistical significance ( P = 0.004). There was no significant difference in the stimulation group and the stimulation group, and the difference was not statistically significant. to learn from one's The difference was statistically significant (P = 0.323) in the control group (P = 0.323). Conclusion 1. Chronic unpredictability stress The expression of MKP-1 in the hippocampus of the rat hippocampus increased with the level of histone H3 in the promoter region, and may be involved in the pathogenesis of depression.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.4
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