Effects Of Huazhuojiedushuganfang On Behavior And Expression Of TPH2 In The Brain Of Depression Model Mice

Objective: To observe the effect of Huazhuojiedushuganfang on the behavior of the mouse model,the model of depression mouse was established by solitary feeding and chronic unpredictable mild stress(CUMS).Additionally,the curative effect of Huazhuojiedushuganfang were analyzed with immnohistochemical staining and the distribution and the expression of tryptophan hydroxylase-2(TPH2)in the mouse raphe nucleus.We also explore the mechanism of Huazhuojiedushuganfang in preventing depression,and then providing experimental evidence for the Treatment of depression.Experimental methods: After a week of adaptive feeding,72 ICR male mice were measured the preference ratio of sugar water.According to the preference ratio of sugar water,mice were divided into six groups,the normal group,model group,baicalin group,fluoxetine group,and Huazhuojiedushuganfang group(low-dose group,medium-dose group and high-dose group)with 12 mice in each group,the average of sugar water preference ratio of each group was similar.After these treatments,behavior of mice were analyzed with novelty suppressed feeding test,Forced swimming test,sugar water preference test,tail suspension test.the immunohistochemical staining showed the positive neurons of tryptophan hydroxylase 2(TPH2).The number of TPH2 positive neurons in the nucleus of the nucleus and the nucleus of the brain bridge were counted.The data were analyzed by one-way anova method,and the difference between the two groups was statistically significant using the Newman-Keuls test.Results: 1.Behavioral test results show: There was a significant difference between the overall mean value of sugar water preference in each group(p <0.05).The sugar water preference in the model group was lower than that in the normal group,and the difference was significantly significant(p< 0.05);There was a significant difference between the overall mean of the fixed time of the mouse tail(p<0.01);In the novel inhibitory feeding experiment,the feeding incubation period of the model group was significantly longer than that of the normal group(p<0.01);In the forced swimming test,the model group swimming incubation period significantly longer than the model group and the medication group(p < 0.05).2.Immunohistochemical test results show that: There were significantly fewer TPH2 positive neurons in the model group(DRD)than the normal group and turbidified detoxification,and the dose group(p<0.05);The TPH2 positive neurons in the ventral side of the dorsal core(DRV)were significantly less than the normal group(P<0.05);The TPH2 positive neurons of the dorsal nuclear beam shoulder(DRI)were significantly less than the normal group(p<0.05);The TPH2 positive neurons in the caudal end of the middle seam(CLi)were significantly less than the normal group(p<0.05);The TPH2 positive neurons in the median nucleus(Mn R)of the middle seam(Mn R)were higher than the normal group(p>0.05)Compared with the model group;Near the center of the raphe nucleus(PMn R)TPH2 positive neurons of model group was significantly less than the fluoxetine group and turbidity of liver detoxification,high dose group(p < 0.05);The TPH2 positive neurons were significantly lower than that of the turbidized antivirus(p<0.05);The TPH2 positive neurons in the inner nucleus of the model group were significantly less than that of the TPH2 positive neurons and the low,medium and high dose group(p<0.05).Discussion: 1.Novel and inhibition of feeding the incubation period,the sugar water preference,tail suspension fixed time and the tail suspension without moving the incubation period,forced swimming and fixed the incubation period and the fixed total time indexes reflect this research adopts the improved chronic mild stress(CUMS)unpredictability normal group take close cage,other groups of single cage measures,make the depression model mice.2.Fluoxetine,baicalin,turbidized and detoxifying liver can improve the depressive behavior caused by the combination of stress and isolation to a certain extent.3.Turbidity detoxification can significantly improve depressive behavior in depression model mice.4.In model group,the TPH2 positive neurons decreased in raphe nucleus,pons,covered the nucleus reticular nucleus and inside of the pons tegmental nucleus and other brain regions.positive neuron increased in the turbidity liver detoxification corresponding regions of the party group increased markedly positive neurons or have increasing trend,indicating that the turbidity liver detoxification by promoting TPH2 expression,the increased multiply 5-HT synthesis that is an important way for the prevention and treatment of depression.5.depression-like behavior shown differences sensitivity of to the three drugs.Moreover,DRD,DRV,DRI,pons tegmental reticular nucleus,pons,covered the medial nuclear TPH2 positive neurons in the brain,such as fluoxetine group,baicalin,shown no significant difference,suggesting that antidepressant effect of fluoxetine and baicalin has no relationship with TPH2 expression in corresponding regions and indicating the mechanism of action of antidepressant is different compared with turbidity liver detoxification.