The Effect Of Shuyu Capsule And Bupleurum Extract Serum On GABA_BR₂-mediated Signal Pathway Of ERK AndAC/CAMP/CREB In Primary Cultured Rat Hippocampus Neurons

Objective:Our aim was to investigate the protein levels of γ-aminobutyric acid B2(GABABR2) receptor in rat hippocampus neurons with the model of rat depressive mood serum i nterventi on, to study the correcti ng f uncti on of Shu Yu capsule and Bupleurum extract which gives a preliminary discussion on the pathogenesis of depression emotion.as well as studies the effects of Shuyu capsule; we activated this receptor using GABAbR2specificity agonists to detect contain medici ne serum was how to activate the effector level of changes of downstream, Our aim was to explore the transduction mechanism of ShuYu capsule on GABAbR2downstream signal pathway.Methods1、we used the chronic mild stress(CMS)to establish the depression emotion model i n rats, and then the rats were eval uated by sucrose i ntake、the body weight and open-field from beginning to the end for the model.Serum of rats in each group was collected and then added into the the medium of vitro cultured hippocampi primary neurons.At last the expression of GABABR2and P-ERK1/2were detected with VVestern Blot respectively.2, Combining with serum pharmacology and cell culture technology, we activated this receptor using GABABR2specificity agonists, and observed the change of the cAMP level and the expression of P-ERK1/2and P-CREB.Results1、Compared with the normal group,the weight, the total score of open field test and the sucrose intake of preference test of rats in depression emotion model group lessened significantly (P<0.01); On the other hand.compared with the depression emotion model group, the weight.the total score of open field test and the sucrose intake of preference test of rats in Shuyu capsule,bupleurum and fluoxetine treatment groups increased significantly(P<0.05or P<0.01). Western Blot demonstrated that the serum of rats in model group obviously increased the protein levels of GABABR2and P-ERK1/2(P<0.01), compared with the normal control group;While serum of rats in the Shuyu capsule and fluoxetine treatment groups markedly decreased the protein levels of GABABR2and P-ERK1/2(P<0.05), the bupleurum group had no difference with the model group.but the trend has improved.2、After acting the hippocampal neurons GABABR2by the specific agonists, The results found that, compared with the blank group, the cAM P level and the expression of P-ERK1/2and P-CREB of the agonist group increased significantly (P<0.01). compared with the agoni st group, the Shuyu capsul e group and the antagonists group decreased the levels of cAM P (P<0.01), the bupleurum group had no difference with the agonist group, but the trend has improved; the antagonists group decreased the expression of P-ERK1/2(P<0.01), the Shuyu group without significantly reduced, the Shuyu capsule group and the antagonists group decreased the expression of P-CREB, the bupleurum group had no difference with the agonist group (P<0.05, P<0.01), the bupleurum group had no difference with the agonist group.Conclusion:1、The model rats with depression emotion can be prepared successfully by using chronicity mild stimulus (CMS),and they can be evaluated effectively by sucrose preference test, open field test and macro-behavior observation.2^The model of rat depressive mood serum can increase the protein levels of GABABR2in the hippocampal neurons, GABABR2may have close relationship with depression emotion.and probably is one of the centre pathogenesis of depression emotion.3> The experimental results of western blot and immunofluorescence show that, the regulating liver formula ShuYu capsule and active ingredient can efficiently correct the abnormal expression of GABABR2, as a result to reach the therapeutic aim, and this drug can reduce the signal pathway of GABABR2in the hippocampus neurons,so that may be one of the mechanisms of the drug treatment for depression emotion.