逍遥散干预PolyI:C诱导小鼠肝郁证的作用研究

发布时间：2018-05-08 23:01

本文选题：肝郁证 + PolyI:C　；参考：《南方医科大学》2017年硕士论文

【摘要】：目的建立PolyI:C诱导小鼠肝郁证模型,探讨逍遥散对PolyI:C诱导小鼠肝郁模型的作用及其作用机制。方法1.PolyI:C诱导小鼠肝郁证模型的建立:第一批:将40只C57小鼠随机分为4组,对照组、3mg/kg PolyI:C组、6mg/kgPolyI:C组、12mg/kgPolyI:C组。除对照组外,其余三组予腹腔注射相应浓度的PolyI:C溶液,用药1天。第二批:将20只C57小鼠随机分为2组,对照组、12 mg/kg Poly I:C造模组。造模组予腹腔注射12 mg/kg Poly I:C溶液,对照组予腹腔注射生理盐水,连续3周。第三批:将60只C57小鼠随机分为6组,对照组、模型组、逍遥散低、中、高剂量组、艾司西酞普兰组。除对照组外,其余三组予腹腔注射PolyI:C,在造模前30分钟予灌胃给药,每天1次,连续3周。2.动物模型的评价在造模后运用旷场实验、糖水偏好实验、强迫游泳实验、悬尾实验等评价行为学指标。3.动物组织样品的采集与检测在完成造模和行为学实验后,将小鼠麻醉,经心脏采血后进行灌注,并采集小鼠大脑组织,用于免疫荧光分析、免疫印迹和酶联免疫测定。结果1.高剂量PolyI:C连续3周造模能成功建立小鼠肝郁证模型。与对照组比较,12 mg/kg PolyI(C组的小鼠中央区域停留时间明显减少(P0.05),糖水偏好分数明显下降(P0.05),悬尾实验和强迫游泳实验的不动时间延长(P0.05)。2.逍遥散能改善PolyI:C诱导的小鼠抑郁样行为。与模型组比较,高剂量逍遥散组糖水偏好分数显著升高(P0.05),悬尾实验和强迫游泳实验的不动时间明显减少(P0.05)。3.逍遥散能调节PolyI:C诱导的前额皮质、VTA、NAc炎症细胞因子。与模型组比较,高剂量逍遥散和西酞普兰均能降低前额皮质显著升高的IL-1β、IL-6及TNF-α的水平(P0.05);高剂量逍遥散可以降低VTA显著升高的IL-6水平(P0.05),而西酞普兰能下调IL-1β、IL-6及TNF-α的水平(P0.05);高剂量逍遥散和西酞普兰均能降低NAc显著升高的IL-6及TNF-α水平(P0.05)。4.逍遥散能下调PolyI:C造模后小鼠前额皮质、VTA、NAc升高的小胶质细胞数。与模型组比较,高剂量逍遥散和西酞普兰能下调前额皮质中显著升高的小胶质细胞数(P0.05);高剂量逍遥散和艾司西酞普兰能下调VTA显著升高的小胶质细胞数(P0.05),西酞普兰还能下调VTA升高的星型胶质细胞数(P0.05);高剂量逍遥散和西酞普兰能下调NAc显著升高的小胶质细胞数(P0.05)。5.逍遥散改善PolyI:C诱导肝郁证的作用可能与调节IDO、p38、GCH1蛋白的表达有关。在前额皮质中,IDO和p38表达升高,逍遥散和西酞普兰能下调其表达。在V,TA中,GCH1表达升高,逍遥散和西酞普兰均能下调其表达。在NAc中,p38、GCH1表达升高,逍遥散能下调GCH1表达,西酞普兰能下调p38、GCH1表达。结论逍遥散对PolyI:C诱导的肝郁证小鼠具有预防性抗抑郁和抗神经炎症作用。长期使用PolyI:C能诱导小鼠大脑前额皮质、VTA、NAc脑区产生神经炎症反应,这些改变能被逍遥散所改善。逍遥散对PolyI:C诱导肝郁证的抗抑郁作用可能与调节IDO、p38、GCH1蛋白表达有关。
[Abstract]:Objective to establish the model of liver depression induced by PolyI:C in mice and to explore the effect and mechanism of Xiaoyao Powder on the model of liver depression induced by PolyI:C in mice. Methods the model of liver depression induced by 1.PolyI:C in mice was established. The first batch: 40 C57 mice were randomly divided into 4 groups. The control group was divided into 3 mg / kg PolyI:C group and 6 mg / kg polyI: C group with 12 mg / kg polyi: C group. In addition to the control group, the other three groups were given intraperitoneal injection of corresponding concentration of PolyI:C solution for 1 day. The second batch: 20 C57 mice were randomly divided into two groups, the control group was treated with 12 mg/kg Poly I: C model. The model group was intraperitoneally injected with 12 mg/kg Poly I: C solution and the control group was treated with saline for 3 weeks. The third batch: 60 C57 mice were randomly divided into 6 groups: control group, model group, low, medium, high dose group and Escitalopram group. In addition to the control group, the other three groups were intraperitoneally injected with PolyI: C and given intragastric administration 30 minutes before modeling, once a day for 3 weeks. The animal model was evaluated with open field experiment, sugar water preference test, forced swimming test, tail suspension test and so on. The collection and detection of animal tissue samples after the completion of model and behavioral experiments, the mice were anesthetized and perfused with blood collected from the heart. The brain tissues of the mice were collected for immunofluorescence analysis, immunoblotting and enzyme-linked immunosorbent assay (Elisa). Result 1. The model of liver depression in mice was successfully established by high dose PolyI:C for 3 weeks. Compared with the control group, the retention time in the central area of the mice in 12 mg/kg PolyI(C group decreased significantly, the preference fraction of sugar water decreased significantly, and the immobility time of tail suspension test and forced swimming test was prolonged (P0.05 路2). Xiaoyao Powder can improve the depressive behavior induced by PolyI:C in mice. Compared with the model group, the sugar water preference fraction in the high dose Xiaoyao powder group was significantly higher than that in the model group, and the immobility time of the tail suspension test and forced swimming test was significantly decreased. Xiaoyao Powder can regulate the inflammatory cytokines of VTA-NAc in prefrontal cortex induced by PolyI:C. Compared with the model group, Both high dose Xiaoyao Powder and citalopram could decrease the levels of IL-1 尾 -IL-6 and TNF- 伪 in the prefrontal cortex, and high dose Xiaoyao Powder could decrease the IL-6 level of VTA, while citalopram could down-regulate the levels of IL-1 尾 -IL-6 and TNF- 伪 (P0.05), and high dose Xiaoyao San could decrease the level of IL-1 尾 -IL-6 and TNF- 伪 (P0.05). Both telecontrol and citalopram could decrease the levels of IL-6 and TNF- 伪, which were significantly increased in NAc. Xiaoyao Powder can down-regulate the increased number of microglia in prefrontal cortex of mice after PolyI:C model. Compared with the model group, High dose Xiaoyao San and citalopram could down-regulate the significantly increased microglial cell count in prefrontal cortex (P0.05), high dose Xiaoyao Powder and Escitalopram could down-regulate the number of microglia cells significantly increased in VTA, and citalopram could also down-regulate the increase of VTA. High dose of Xiaoyao San and citalopram could down-regulate the number of microglial cells significantly increased in NAc. The effect of Xiaoyao Powder on PolyI:C induced liver depression may be related to the regulation of the expression of GCH1 protein in IDOP38. In prefrontal cortex, the expression of IDO and p38 increased, but Xiaoyao San and citalopram could down-regulate the expression of IDO and p38. The expression of GCH1 in VTA was increased, and both Xiaoyao San and citalopram could down-regulate its expression. In NAc, the expression of p38 GCH1 was increased, the expression of GCH1 was down-regulated by Xiaoyao San, and the expression of p38 GCH1 was down-regulated by citalopram. Conclusion Xiaoyao Powder has preventive antidepressant and anti-inflammatory effects on PolyI:C induced liver depression mice. Long-term use of PolyI:C could induce neuroinflammatory responses in the prefrontal cortex of mice, and these changes could be improved by Xiaoyao San. The antidepressant effect of Xiaoyao Powder on PolyI:C induced liver depression may be related to regulating the expression of GCH1 protein in IDO p38.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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