【作者】 徐明婧；

【导师】 张惠云；

【作者基本信息】 山东中医药大学 ， 中药学， 2011， 硕士

【摘要】 目的:检测抑郁情绪模型大鼠脑区额叶皮质、顶叶皮质、海马和下丘脑相关区域内5-HT_3B受体(5-HT_3BR)蛋白分布、表达的变化,初步探讨抑郁情绪发病的中枢机制,并研究中药舒郁胶囊对该病证的干预作用。方法:筛选大鼠进入实验,采用慢性温和应激法（CMS）复制抑郁情绪大鼠模型,以糖水偏好实验、旷场试验和宏观行为表现评价模型,以西药氟西汀为阳性药,采用免疫荧光法检测正常对照组、抑郁情绪模型组、舒郁胶囊给药组和氟西汀西药给药组大鼠额叶皮质、顶叶皮质、海马和下丘脑中5-HT_3BR的定位分布,并定量检测海马与下丘脑中5-HT_3BR的表达。结果:与正常组大鼠相比,模型组大鼠体重显著减轻（P<0.01）,旷场实验水平得分、垂直得分、旷场实验总分均显著减少（P<0.01）,糖水偏好实验糖水摄入量显著减少（P<0.01）,大鼠表现出精神萎靡,眼神呆滞,对外界刺激不敏感的状态;而造模并给予舒郁胶囊和氟西汀的大鼠较模型组大鼠体重增长显著（P<0.05）,旷场实验三项得分均显著增加（P<0.01）,糖水偏好实验中饮糖水量显著增加（P<0.01）,但两组之间都没有显著性差异（P>0.05）。免疫荧光结果显示,四组大鼠额叶皮质、顶叶皮质、海马、下丘脑中5-HT_3BR的定位分布无显著性差异。与正常组大鼠相比,模型组大鼠海马CA1、CA3与下丘脑VMH区5-HT_3BR平均荧光强度（MFI）显著升高（P<0.05或P<0.01）,额叶皮质、顶叶皮质5-HT_3BR有升高的可能。造模并给予舒郁胶囊和氟西汀治疗的大鼠与模型组相比,海马CA1、CA3与下丘脑VMH区5-HT_3BR平均荧光强度（MFI）显著降低（P<0.01）,两组间没有显著性差异（P>0.05）,额叶皮质、顶叶皮质5-HT_3BR有降低的可能。结论:采用慢性温和应激法可以成功制备抑郁情绪大鼠模型,糖水偏好实验、旷场试验结合宏观行为药理学可有效评价模型。大鼠额叶皮质、顶叶皮质、海马CA1、CA3、下丘脑VMH区5-HT_3BR蛋白表达升高可能与抑郁情绪关系密切,可能是抑郁情绪发病的中枢机制之一;中药舒郁胶囊可以有效纠正模型大鼠上述脑区5-HT_3BR蛋白表达异常升高的状态,可能是该药治疗抑郁情绪的机制之一。更多还原

【Abstract】 Objective:Our aim is to detect the relationship between protein location & expression levels of 5-HT_3BR in correlated region offrontal cortex , parietal cortex ,hippocampus & hypothalamus of depression emotion model rats, which gives a preliminary discussion on the pathogenesis of depression emotion,as well as studies the effects of Shuyu capsule.Methods:After being chosen,depression emotion model rats were prepared by using chronicity mild stimulus（CMS）method.And then the rats were evaluated by sucrose preference test,open field test and macro-behavior observation.We used fluoxetine as masculine medicine, detected the 5-HT_3BR protein location in parietal cortex, frontal cortex, hippocampus and hypothalamus of rats in the groups including the normal group, depression emotion model group,and the two treatment groups by using immunofluorescence（IF）, and detected the mean fluorescence intensity （MFI） in CA1、CA3 of hippocampus and in ventromedial hypothalamic area（VMH）of rats.Results: Compared with the normal group,the weight of rats in depression emotion model group lessened significantly（P<0.01）, the crossing score,rearing score and total score of open field test all decreased obviously（P<0.01） and the sucrose intake of preference test decreased obviously（P<0.01）.Meanwhile, the rats showed listlessness,dull in eyes and insensitive to surrounding stimulation.On the other hand,compared with the depression emotion model group,the weight of rats in Shuyu capsule and fluoxetine treatment groups increased significantly（P<0.05）, the open field scores all increased remarkably（P<0.01）,and the sucrose intake of preference test increased obviously（P<0.01）,and there was no statistical differences in the two treatment groups（P>0.05）.IF demonstrated that there was no noticeable differences in location of 5-HT_3BR infrontal cortex, parietal cortex, hippocampus and hypothalamus of rats in four groups.Compared with the normal group,the numerical value of MFI of 5-HT_3BR in depression emotion model group increased markedly （P<0.05 or P<0.01）in CA1、CA3 of hippocampus and in VMH, there maybe a elevated possibility of 5-HT_3BR in frontal cortex and parietal cortex. Compared with the depression emotion model group, the numerical value of MFI of 5-HT_3BR in Shuyu capsule and fluoxetine treatment groups decreased markedly （P<0.05）in CA1 of hippocampus and in VMH, there was no statistical differences in the two treatment groups（P>0.05）, and there maybe a reducing possibility of 5-HT_3BR in frontal cortex and parietal cortex.Conclusion: The model rats with depression emotion can be prepared successfully by using chronicity mild stimulus（CMS）,and they can be evaluated effectively by sucrose preference test,open field test and macro-behavior observation.The high expressions of the5-HT_3BR protein in frontal cortex, parietal cortex, CA1、CA3 of hippocampus and in VMH may have close relationship with depression emotion,and probably is one of the centre pathogenesis of depression emotion.The Shuyu capsule can correct the high expression of 5-HT_3BR protein of model rats in above brain regions, which may be one of the mechanism to treat depression emotion.更多还原