The Mechanism Of The Dorsal Raphe Nucleus GABABR1 Mediated GABA Regulating Prefrontal Cortex Serotonin Level In Anger-out And Anger-in Male Rats

Objectives To explore the mechanism of the Dorsal Raphe Nucleus(DRN)GABABR1(GB1)mediated GABA regulating Prefrontal Cortex(PFC)serotonin(5-HT)level in Anger-out and Anger-in male rats.To further explore the difference of the anger-out and anger-in and the intervention mechanism of Baixiangdan Capsule(BXD)and Shuyu Capsules(SY).Methods A rat model of anger induced by social isolation combined with residential invasion.With behavioral evaluation to distinguish Anger-out and Anger-in model rats.BXD and SY were separately used as intervention medicine of Anger-out and Anger-in rats.Based on this,application of Enzyme Linked Immunosorbent Assay(ELISA)technology to detect GABA content in the DRN and5-HT content in the PFC after liver regulating prescription effecting in the different time-Histories.Immunofluorescence double labeling technique(IFDL)was used to detect the coexpression of 5-HT and GB1 in the DRN.Immunoelectronmicroscopy was used to detect the coexpression of 5-HT and GB1 in the DRN by subcellular level.Finally,baclofen,a GB1 specific agonist,and CGP35348,a GB1 specific inhibitor,was injected into the DRN by the stereotaxic microinjection technique.Then PFC 5-HT level were detected via ELISA.Results(1)High and low Aggressive Behavior Scores(ABS)and Total Distance(TD)were significantly distinguished in Anger-out Group and Anger-in Group after modeling(P<0.05).At the same time,there was a significant decrease in Sugar Preference Coefficient(SPC)and Relative Sugar Yield(RSY)(P<0.05).After corresponding administration,ABS and TD of BXD Group were significantly lower than Anger-out model Group(P<0.05),on the contrary,ABS and TD of SY Group were significantly higher than Anger-in model Group(P<0.05),SPC and RSY of BXD Group and SY Group were significantly higher than Anger-out model Group and Anger-in model Group(P<0.05).(2)The PFC 5-HT content of Anger-out model Group and Anger-in model Group were lower than Control Group and always maintained at a low level.After corresponding administration,with the increase of medication duration,the content of 5-HT increased in PFC,and gradually corrected the abnormal state,which was significantly higher than model Group(P<0.05)at seventh days.On the contrary,the DRN GABA content of Anger-out model Group and Anger-in model Group were higher than Control Group and always maintained at a high level.After corresponding administration,with the increase of medication duration,the content of GABA decreased in the DRN,and gradually corrected the abnormal state,which was significantly lower than model Group(P<0.05)at seventh days.(3)IFDL result showed that a large number of 5-HT positive staining cells(red)in DRN also showed positive staining of GB1(green),that is,the expression of 5-HT neurons in DRN could be coexpressed with GB1.Semi-quantitative analysis showed that GB1 expression in DRN of Anger-out model Group and Anger-in model Group were significantly higher than control group(P<0.05),and Anger-in model Group was significantly higher than Anger-out model Group(P<0.05).After corresponding administration,it was decreased significantly(P<0.05),reached the normal level(P>0.05).On the contrary,5-HT expression in DRN of Anger-out model Group and Anger-in model Group were significantly lower than control group(P<0.05),and Anger-in model Group was significantly lower than Anger-out model Group(P<0.05).After corresponding administration,it was increased significantly(P<0.05),reached the normal level(P>0.05).(4)The results of immunoelectron microscopy showed that GB1 particles were located in the synaptic membrane containing 5-HT particles.(5)Injected saline as control,after injecting baclofen into DRN,the PFC 5-HT content were significantly decreased(P<0.05).On the contrary,after injection of CGP35348,the PFC 5-HT content were significantly increased(P<0.05).Conclusions The method of social isolation combined with residential invasion can successfully create a stable and scientific Anger-out and Anger-in model.GABA significantly increased in the DRN and GB1 significantly increased in the DRN 5-HT neurons binding,further mediated synaptic inhibition effect,thereby reducing the 5-HT in DRN,resulting in 5-HT level of PFC decreased significantly.Therefore,GB1 in DRN mediated GABA regulation of PFC 5-HT level is one of the mechanisms of anger-out and anger-in.The different degree of abnormal increase of GB1 expression in the DRN can be mediated by GABA and the degree of 5-HT reduction in the 5-HT neurons of the DRN is also different,which are different in pathogenesis of anger-out and anger-in.BXD and SY can effectively correct the abnormal changes,and optimal action time was continuous medication for 7 days.