非甾体抗炎药尼美舒利对CUMS大鼠抑郁行为的影响
发布时间：2018-11-26 10:09
【摘要】：目的：本实验采用慢性不可预见性温和应激(chronic unpredictablemild stress，CUMS)建立大鼠抑郁模型，观察尼美舒利对慢性应激大鼠抑郁行为的影响。采用比较蛋白质组学研究策略，运用ITRAQ联合LC-MS/MS技术检测正常组及各用药组大鼠海马组织与模型组相比较所得的差异蛋白，以期从蛋白质组学角度阐述尼美舒利对抑郁大鼠的作用机制并希望找到新的作用靶点。 方法：（1）120只成年雄性SD大鼠，随机分为模型组、正常组、阳性对照组和尼美舒利3,6和12mgkg-1组。正常组不进行任何刺激，其余各组采用慢性不可预见性轻度应激方法制备大鼠抑郁模型。造模完成后，分别给予尼美舒利3mgkg-1、6mgkg-1和12mgkg-1，阳性对照组给予氟西汀3mgkg-1，正常组和模型组给予浓度为0.5%等体积的羧甲基纤维素钠，每天灌胃1次,连续21d。 （2）通过高架十字迷宫、旷场实验和强迫游泳试验，检测大鼠行为学改变；通过组织病理切片HE染色，观察大鼠海马神经细胞的形态变化，通过免疫组织化学染色，观察大鼠海马神经细胞SYN、NGF、BDNF、5-HT1A受体蛋白表达；通过酶联免疫吸附法，检测大鼠海马组织中的IL-1β、IL-4、IL-6、IL-10和TNF-α的含量变化。 （3）采用比较蛋白质组学研究策略，ITRAQ联合LC-MS/MS筛选大鼠海马组织表达有差异的蛋白质。 结果：（1）模型组大鼠进入高架十字迷宫进入总次数和开臂次数及停留时间百分率均较正常组减少；强迫游泳实验中，静止不动的时间相比正常组增加；旷场实验中，垂直活动次数和水平活动次数相比正常组减少。模型组大鼠海马组织细胞的IL-1β、TNF-α和IL-6含量升高，IL-4和IL-10含量降低；海马组织中神经细胞出现细胞丢失、核固缩，SYN、NGF、BDNF及5-HT1A受体蛋白表达降低。 （2）尼美舒利各剂量组能明显改善大鼠的抑郁行为，降低海马组织细胞IL-1β、TNF-α和IL-6含量，升高IL-4、 IL-10含量。减轻慢性应激大鼠海马神经的损伤，阻遏海马神经细胞BDNF、NGF、SYN、5-HT1A表达下调。 (3)与正常组比较，模型组筛选出54个差异蛋白质，41个高表达，13个低表达。从尼美舒利3mgkg-1组筛选到75个差异蛋白，55个高表达，20个低表达。从尼美舒利6mgkg-1组筛选到40个差异蛋白，31个高表达，9个低表达。从尼美舒利12mgkg-1组筛选到42个差异蛋白，12个高表达，30个低表达。从氟西汀组筛选到70个差异蛋白，57个高表达，13个低表达。并且发现Tyrosine-protein kinase receptor（酪氨酸蛋白激酶受体），Sodium-and chloride-dependent GABA transporter3（SLC6A11）这两个差异蛋白在正常组，氟西汀组，尼美舒利3mgkg-1、6mgkg-1和12mgkg-1组筛选出的差异蛋白中均有出现，uniport数据库分析其功能主要与树突细胞因子的产生，，炎症应答，细胞增殖，神经的可塑性，脑发育，神经递质的运输等功能密切相关。这两个蛋白质在忧郁症的发病、预后起到什么作用需要进一步深入研究。 结论：尼美舒利各剂量组对CUMS大鼠的抑郁行为有明显的改善作用，其作用机制可能与抑制炎症因子的释放,增加神经可塑性有关。比较蛋白质组学结果表明海马组织蛋白质组发生的这种复杂变化对抑郁症的发病及预后起到十分重要的作用，两个共同蛋白质存在正常组与给药组表明炎性因子及神经可塑性相关蛋白可能是潜在的抗抑郁药物靶点。
[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.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
本文编号：2358251
[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.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
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本文编号：2358251
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