| Objectives:To explore the mechanism as well as the similarities and differences of 5-HT levels in hypothalamus regulated by Dorsal Raphe Nucleus(DRN)GABABR1 mediated GABA in anger-in and anger-out male rats,so as to provide scientific basis for the effective prevention and cure of the related disease syndrome induced by anger of Gan-regulating medicine.Methods:The anger-out and anger-in male rats model was established by social isolation combined with resident-intruder stress,and mainly tested by aggressive behavior scores(ABS).The model was distinguished by open field test and sucrose preference test and intervened by corresponding Gan-regulating medicine.On this basis,ELISA was used to detect 5-HT level in Hypothalamus and GABA level in DRN in different days of application(Administration by Gan-regulating medicine after 0,1,3,5,7 days);Immunofluorescence(IF)double labeling technique was used to detect the common expression of 5-HT and GABABR1 in the DRN as well as the intensity of IF,and detection of synaptic connections between 5-HT and GABABR1 in DRN was by immunoelectron microscopy double labeling technique;Determination of 5-HT content in hypothalamus was by ELISA after stereotactic injection of GABABR1 agonist Baclofen or inhibitor CGP35348.Results:(1)Anger-out and anger-in rats acted different ABS and open field distance(OFT)after modeling.After intervence,ABS of anger-out BXD was significantly lower than anger-out model group(P<0.05),ABS of anger-in SY was significantly higher than anger-in model group(P<0.05),and when two drug group levels were restored to the control group level(P>0.05),OFT also showed the same trend.(2)5-HT level in hypothalamus in the model group was significantly lower than normal group(P<0.05),and the GABA content of DRN was significantly higher than the normal group(P<0.05),and there was significant difference between the two model groups as well(P<0.05).After intervence of Gan-regulating medicine,with the increase of medication days,the 5-HT level in the hypothalamus of the treatment group was gradually increased and significantly higher than the model rats(P<0.05)at the seventh day,and was restored to normal(P>0.05);GABA level was contrary at the same time.(3)Determinations of IF double labeling technique showed 5-HT neurons could be co-expressed with GABABR1 in DRN.Determination of fluorescence semi-quantitative showed: compared with the control group,the GABABR1 levels in anger-out and anger-in model group were significantly increased(P<0.05),and anger-in model group was more higher(P<0.05),but after treatment the level was significantly decreased(P<0.05)and then restored to normal(P>0.05);5-HT was just contrary.(4)The determination of immunoelectron microscopy from the rat sections showed GABABR1 is located at the synaptic membrane containing 5-HT.(5)Compared with injection of normal saline,the 5-HT content in hypothalamus of rats in each group was significantly decreased after injection of GABABR1 agonist Baclofen(P<0.05);on the contrary,after injection of CGP35348,5-HT content in the hypothalamus of rats in each group was significantly increased(P<0.05).Conclusion:Social isolation combined with resident-intruder stress could be copied to establish anger-out and anger-in rats model and the method was stable.As two forms of anger,anger-out and anger-in can both increase the GABA and GABABR1 expression in DRN,and then cause the decrease of 5-HT release in DRN and significantly decrease of 5-HT content in hypothalamus.Relevant gan-regulating medicine can be applied to these targets and effectively cure disease,which is same between anger-out and anger-in.Different levels of abnormal increase of GABABR1 expression in DRN can lead to different releases of 5-HT mediated by GABA,where anger-out differ from anger-in. |
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