Proteomic Analysis Of Prefrontal Cortex In CUMS Depressive Rat Model Treated With Acupuncture Combined With Paroxetine By ITRAQ

ObjectiveThis research was carried out to screen prefrontal cortex(PFC)'s differentially expressed proteins of depressive rat models after stimulated by chronic unpredictable mild stress(CUMS),based on Isobaric Tags for Relative and Absolute Quantitation(iTRAQ),thus to investigate the rapidly therapeutic mechanisms of electroacupuncture combined Paroxetine.MethodsIn this study,CUMS was used for 21 days to induced depressive rat models,which included stimulations such as water deprivation,fasting,swimming in cold water and swimming in warm water.After 21 days' CUMS stimulations,successfully depressive rat models were given electroacupuncture combined Paroxetine,electroacupuncture or Paroxetine treatments.Behavioral tests such as weight monitor,sucrose preference test(SPT)and open field test(OFT)were used to evaluate antidepressant effect of treatments,while iTRAQ was applied to identify differentially expressed proteins in PFC,thus to reveal potential proteins participated in the pathogenesis of depression and rapidly therapeutic mechanisms of electroacupuncture combined Paroxetine.Results1.After CUMS stimulation,weight,SPT and OFT of depressive rats was decreased.After 1 week's treatment,weight,consumption of sucrose water,sucrose'preference,time in center zone and motion distance of depressive rats were increased,among which electroacupuncture combined Paroxetine treatment exhibited the most significant effect.Above-mentioned indexes were increased significantly in electroacupuncture combined Paroxetine treatment group than control group.And time in center in electroacupuncture combined Paroxetine group was increased significantly than electroacupuncture group and Paroxetine group.Moreover,there was no significant difference in consumption of sucrose water,sucrose' preference and time in center zone between electroacupuncture combined Paroxetine group and normal group.2.170 proteins of PFC were screened by iTRAQ.Bioinformatics analysis results of differentially expressed proteins indicated that molecular functions of those proteins relevant to binding functions and enzymatic activity primarily,and biological processes were mainly about maintaining structural integrity of neurons,signal transduction,transport and metabolism,while their pathways included endocytosis,dopaminergic synapse and serotonergic synapse primarily.Furthermore,interactive networks of differentially expressed proteins centre on dopamine metabolism and regulations of immunological function.3.Verification results of western blot(WB)showed that expressions of protein Bassoon,glutamate receptor 3(GluR3),Aromatic-L-amino-acid decarboxylase(DDC)and Tyrosine 3-monooxygenase(TH)were decreased in control group,with Bassoon exhibited significant reduction.After 1 week's treatment,expressions of Bassoon,GluR3,DDC and TH were increased,with DDC increased significantly in electroacupuncture combined Paroxetine group and TH increased significantly in electroacupuncture group when compared with control group.ConclusionElectroacupuncture combined with Paroxetine won a rapid effect after one week's treatment.Pathogenesis of depression is concerned with reductions of endocytosis,excitatory receptor and monoamine neurotransmitters likely.Rapidly therapeutic mechanisms of electroacupuncture combined Paroxetine treatment may concerning regulations of endocytosis,increase of excitatory receptor and monoamine neurotransmitters.Therefore,multiple proteins,targets and factors are involved in pathogenesis of depression and rapidly therapeutic mechanisms of electroacupuncture combined Paroxetine.