逍遥散对慢性心理应激大鼠行为学及代谢组学的影响
发布时间：2019-05-20 17:04
【摘要】：目的:观察慢性束缚应激大鼠粪样中代谢物变化及中药复方逍遥散的调节作用,确定与慢性束缚应激和逍遥散作用密切相关的“代谢组学”小分子标志化合物,以探索逍遥散对慢性心理应激的干预作用及其机制。方法:采用每天3h连续21天束缚制动方式建立慢性应激大鼠模型,以逍遥散为干预用药。72只SD大鼠随机分为正常组、模型组、逍遥散组。模型组和逍遥散组大鼠均每天给予束缚应激,逍遥散组大鼠在束缚应激同时每天灌服逍遥散3.854g/kg,正常组和模型组大鼠灌服0.9%生理盐水。观测各组大鼠的外观、体重及摄食量等变化;采用旷场实验和高架十字迷宫实验观测应激大鼠的自主活动能力和对新异环境的探究能力;采用基于核磁共振氢谱(1H Nuclear Magnetic Resonance,1H NMR)技术的代谢组学方法检测各组大鼠粪样代谢谱变化,通过多元统计分析寻找差异性代谢产物,确定与慢性束缚应激和逍遥散作用机理密切相关的“代谢组学特征”的小分子标志化合物。结果:1外观表现、体重与摄食量变化模型组大鼠的皮毛色泽等外观表现较差,体重和摄食量均较正常组大鼠明显下降(P0.05或P0.01);逍遥散干预大鼠的皮毛色泽等外观表现有所改善,较模型组大鼠体重升高(P0.01),摄食量也有增加趋势,尤其实验第9~15天增加明显。2行为学实验结果2.1旷场实验在实验第7、21天,模型组大鼠5 min总移动路程和在中央区移动路程均较正常组减少,尤其总移动路程下降明显(P0.05或P0.01);逍遥散干预大鼠5 min内总移动路程和中央区移动路程均较模型组大鼠明显增加(P0.05或P0.01)。2.2高架十字迷宫实验在实验第7、21天,模型组大鼠5 min内在开放臂停留时间和进入次数均较正常大鼠减少,尤其实验末第21天变化明显(P0.05或P0.01);模型组大鼠5min内在封闭臂停留时间增加但进入次数减少,在第21天表现也最明显(P0.05或P0.01)。在实验第21天,逍遥散干预大鼠5 min内进入开放臂次数和在开放臂停留时间较模型组大鼠明显增加(P0.05),而在封闭臂停留时间较单纯应激大鼠下降明显(P0.05)但进入次数也显著增加(P0.05)。旷场实验和高架十字迷宫实验说明持续21天的慢性束缚应激引发了大鼠恐惧、焦虑、忧郁等心理反应。3 1H NMR谱结合多元统计分析结果模型组和逍遥散组大鼠粪样中Acetate、Butyrate、Glucose、Propionate、Ribose、Maltose、Valerate、Xylose 8种差异性代谢产物均明显高于正常大鼠(P005或P001);逍遥散组大鼠的8种差异性代谢产物明显高于模型组大鼠(P005或P001)。结论:1逍遥散对慢性心理应激大鼠外观表现、摄食量、体重具有一定的调节作用,并可缓解其恐惧、忧郁等心理反应。2大鼠粪样中以Glucose、Maltose、Ribose、Xylose为主的糖类和以Acetate、Butyrate、Proionate、Valerate为主的短链脂肪酸可作为与慢性心理应激和逍遥散作用关系密切的差异代谢小分子标志化合物。3调节肠道菌群种类及数量从而增加糖类及短链脂肪酸等小分子代谢物形成,是逍遥散干预作用的可能机制之一。
[Abstract]:Objective: To observe the changes of the metabolites in the fecal samples of the rats with chronic binding stress and the regulation of the traditional Chinese medicine, and to determine the small "metabolomics" molecular marker compounds which are closely related to the effects of the chronic binding stress and the dispersing effect. In order to explore the effect and mechanism of the intervention on the chronic psychological stress. Methods: The model of chronic stress rats was established by three-hour continuous 21-day binding braking method. The rats were randomly divided into the normal group, the model group and the scattered group in 72 SD rats. The rats of the model group and the scattered group were given the restraint stress every day, while the rats in the group of the control group and the model group were filled with the saline solution of 3.854g/ kg, and the normal group and the model group were given 0.9% normal saline. The appearance, body weight and food consumption of each group of rats were observed. The self-activity and the exploration ability of the stress rats were observed by open-field experiment and elevated cross-maze test. The nuclear magnetic resonance hydrogen spectrum (1H Nuclear Magnetic Resonance) was used. The metabolomics method of 1H NMR technique was used to detect the changes of the fecal samples in each group, and the small molecular marker compounds of the "metabolomics" which are closely related to the mechanism of chronic binding stress and the mechanism of dispersing are determined by the multivariate statistical analysis to find the difference metabolites. Results: The appearance, body weight and food consumption of rats in the model group showed poor appearance, the body weight and the food consumption decreased significantly (P0.05 or P0.01), and the appearance of the fur and the color of the rats in the rats was improved. In the model group, the weight of the rats was increased (P0.01), and the food consumption was increased, especially in the 9th to 15th day of the experiment. In particular, the total movement distance of the rats was significantly lower than that of the model group (P0.05 or P0.01), and the total movement distance and the movement distance of the central area in the rats were significantly higher than that in the model group (P0.05 or P0.01). The residence time of the open arms and the number of entering times in the model group were less than that in the normal rats (P0.05 or P0.01). The residence time of the closed arm in the model group was increased within 5 minutes, but the number of entering times decreased, and the expression in the 21st day was the most obvious (P0.05 or P0.01). In the 21st day of the experiment, the number of open arms and the number of rats in the model group with the time of the open arms increased significantly (P0.05), while the time of the closed arm was significantly lower than that of the rats with simple stress (P <0.05), but the number of access was also increased significantly (P0.05). The experimental and elevated cross-maze test showed that the chronic binding stress of 21 days resulted in the psychological reaction of the rat's fear, anxiety, and depression. The 1H NMR spectrum combined with the multivariate statistical analysis result model group and the Aetate, Buyrate, Gluconate, Propionate, Ribose, Maltaose, Valeate in the model group and the scattered group rats. The 8 difference metabolites of Xylose were significantly higher than those of normal rats (P005 or P001), and the 8 difference metabolites in the group of Xylose were significantly higher than that in the model group (P005 or P001). Conclusion:1. The effects of 1.1.1. powder on the appearance, food consumption and body weight of the rats with chronic psychological stress can be adjusted, and their fear, depression and other psychological reactions can be relieved. Valerate-based short-chain fatty acid can be used as a marker compound with small molecular marker which is closely related to the effect of chronic psychological stress and dispersion, and the type and quantity of intestinal flora can be adjusted, so as to increase the formation of small molecular metabolites such as saccharides and short-chain fatty acids, It is one of the possible mechanisms of the effect of sporadic intervention.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5
[Abstract]:Objective: To observe the changes of the metabolites in the fecal samples of the rats with chronic binding stress and the regulation of the traditional Chinese medicine, and to determine the small "metabolomics" molecular marker compounds which are closely related to the effects of the chronic binding stress and the dispersing effect. In order to explore the effect and mechanism of the intervention on the chronic psychological stress. Methods: The model of chronic stress rats was established by three-hour continuous 21-day binding braking method. The rats were randomly divided into the normal group, the model group and the scattered group in 72 SD rats. The rats of the model group and the scattered group were given the restraint stress every day, while the rats in the group of the control group and the model group were filled with the saline solution of 3.854g/ kg, and the normal group and the model group were given 0.9% normal saline. The appearance, body weight and food consumption of each group of rats were observed. The self-activity and the exploration ability of the stress rats were observed by open-field experiment and elevated cross-maze test. The nuclear magnetic resonance hydrogen spectrum (1H Nuclear Magnetic Resonance) was used. The metabolomics method of 1H NMR technique was used to detect the changes of the fecal samples in each group, and the small molecular marker compounds of the "metabolomics" which are closely related to the mechanism of chronic binding stress and the mechanism of dispersing are determined by the multivariate statistical analysis to find the difference metabolites. Results: The appearance, body weight and food consumption of rats in the model group showed poor appearance, the body weight and the food consumption decreased significantly (P0.05 or P0.01), and the appearance of the fur and the color of the rats in the rats was improved. In the model group, the weight of the rats was increased (P0.01), and the food consumption was increased, especially in the 9th to 15th day of the experiment. In particular, the total movement distance of the rats was significantly lower than that of the model group (P0.05 or P0.01), and the total movement distance and the movement distance of the central area in the rats were significantly higher than that in the model group (P0.05 or P0.01). The residence time of the open arms and the number of entering times in the model group were less than that in the normal rats (P0.05 or P0.01). The residence time of the closed arm in the model group was increased within 5 minutes, but the number of entering times decreased, and the expression in the 21st day was the most obvious (P0.05 or P0.01). In the 21st day of the experiment, the number of open arms and the number of rats in the model group with the time of the open arms increased significantly (P0.05), while the time of the closed arm was significantly lower than that of the rats with simple stress (P <0.05), but the number of access was also increased significantly (P0.05). The experimental and elevated cross-maze test showed that the chronic binding stress of 21 days resulted in the psychological reaction of the rat's fear, anxiety, and depression. The 1H NMR spectrum combined with the multivariate statistical analysis result model group and the Aetate, Buyrate, Gluconate, Propionate, Ribose, Maltaose, Valeate in the model group and the scattered group rats. The 8 difference metabolites of Xylose were significantly higher than those of normal rats (P005 or P001), and the 8 difference metabolites in the group of Xylose were significantly higher than that in the model group (P005 or P001). Conclusion:1. The effects of 1.1.1. powder on the appearance, food consumption and body weight of the rats with chronic psychological stress can be adjusted, and their fear, depression and other psychological reactions can be relieved. Valerate-based short-chain fatty acid can be used as a marker compound with small molecular marker which is closely related to the effect of chronic psychological stress and dispersion, and the type and quantity of intestinal flora can be adjusted, so as to increase the formation of small molecular metabolites such as saccharides and short-chain fatty acids, It is one of the possible mechanisms of the effect of sporadic intervention.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5
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