金郁欢汤剂灌胃对抑郁症小鼠抑郁样行为的改善作用及其机制

Effect of intragastric administration of Jinyuhuan decoction on depression-like behavior in depressed mice and its mechanism

目的观察金郁欢汤剂灌胃对抑郁症小鼠抑郁样行为的改善作用,并探讨其机制。方法60只ICR小鼠随机分为control组、model组、JYH-L组(金郁欢低剂量)、JYH-H组(金郁欢高剂量)、fluoxetine组(氟西汀),每组12只,其中model组、JYH-L组、JYH-H组、fluoxetine组通过CMUS法建立小鼠抑郁症模型。造模完成后,JYH-L组小鼠给予0.2 g/kg金郁欢汤剂灌胃,JYH-H组给予0.8 g/kg金郁欢汤剂灌胃,fluoxetine组小鼠给予氟西汀悬浮液灌胃,control组和model组小鼠同期等体积生理盐水灌胃,每日1次,连续3周。采用糖水偏嗜实验观察各组小鼠抑郁样行为,以糖水偏嗜度表示。采用Nissl染色法观察小鼠海马区神经细胞形态、尼氏小体染色情况,使用JD-801形态学图像分析系统检测各组小鼠海马CA1区尼氏小体积分光密度(IOD)。采用实时荧光定量PCR法检测各组小鼠海马组织中散乱蛋白1(Dvl1)mRNA,采用Western Blotting法检测各组小鼠海马组织中Dvl1蛋白。结果control组、JYH-L组、JYH-H组、fluoxetine组小鼠糖水偏嗜度均高于model组(P均<0.05),JYH-H组、fluoxetine组小鼠糖水偏嗜度均高于JYH-L组(P均<0.05)。model组、JYH-L组神经细胞排列松散或缺失,尼氏小体被轻微染色;JYH-H组与fluoxetine组海马区神经元细胞排列的更密集、更整齐,神经细胞无核固缩深染现象,尼氏小体染色加深。model组小鼠海马CA1区尼氏小体IOD值低于control组(P<0.05),JYH-H组、fluoxetine组小鼠海马CA1区尼氏小体IOD值均高于model组、JYH-L组(P均<0.05)。model组小鼠海马组织中Dvl1 mRNA和蛋白相对表达量均低于control组(P均<0.05),JYH-H组、fluoxetine组小鼠海马组织中Dvl1 mRNA和蛋白相对表达量均高于model组、JYH-L组(P均<0.05)。结论高剂量(0.8 g/kg)金郁欢汤剂灌胃可改善抑郁症小鼠的抑郁样行为,金郁欢汤剂改善抑郁症小鼠抑郁样行为的作用机制可能与其有效成分可上调海马组织中Dvl1的表达水平有关。

Objective To observe the effect of intragastric administration of Jinyuhuan(JYH) decoction on depression-like behavior in depressed mice and to explore the mechanism.Methods Sixty ICR mice were randomly divided into the control group,model group,JYH-L(low dose) group,JYH-H(high dose) group and fluoxetine(fluoxetine) group,with 12 mice in each.Depressed mouse models were established in the model,JYH-L,JYH-H and fluoxetine groups by CMUS.After completion of molding,mice in the JYH-L group were fed with 0.2 g/kg JYH decoction,mice in the JYH-H group were fed with 0.8 g/kg JYH decoction,fluoxetine group with fluoxetine suspension,and the control and model groups with equal volume of saline,once a day,for 3 weeks.Depression-like behavior in each group was observed using sugar water-biased experiment,which was expressed as sugar water-biased taste.Neurocell morphology and Nisslisome staining in the hippocampus were observed by Nissl staining,and the small volume spectrodensity(IOD) of the hippocampus was detected by using the JD-801 morphological image analysis system.Scattered protein 1(Dvl1) mRNA was detected in each mouse hippocampus by quantitative real-time PCR,and Dvl1 protein by Western blotting.Results The control group,JYH-L group,JYH-H group and fluoxetine group had higher sugar water-biased taste than the model group(all P<0.05),and sugar water-biased taste was higher in the JYH-H group and fluoxetine group than in the JYH-L group(all P<0.05).Neural cells were arranged loosely or absent in the model group and JYH-L group,nissl bodies were stained slightly,more dense and neatly aligned neurons were found in the JYH-H group and fluoxetine group with nerve cells without nuclear fixation and nissl bodies with deeper staining.In the model group,IOD value in the CA1 area was lower than that in the control group(P<0.05),and IOD value in the CA1 area of the JYH-H and fluoxetine groups were higher than those in the model and JYH-L groups(all P<0.05).The relative expression levels of Dvl1 mRNA and protein were lower in the model group than in the control group(all P<0.05).The relative expression levels of Dvl1 mRNA and protein in the hippocampus of mice in the JYH-H and fluoxetine groups were higher than those in the model group and JYH-L group(all P<0.05).Conclusion The intragastric administration of high-dose Jinyuhuan decoction(0.8 g/kg) improves depression-like behavior in depressed mice,and the mechanism of action may be related to the active component of Jinyuhuan decoction to up-regulate Dvl1 expression level in hippocampal tissues.