| OBJECTIVE:Depression is considered as a kind of critical disease which endangers the human being’s health.The pathogenesis of depression is complex and the disease lacks of effective drugs for long-term treatment with low side-effect.Chaihu-Shu-Gan-San(CSGS)is a traditional Chinese medicine prescription(TCMP)with comfirmed effect in treatment of depression in clinic,however,its active basis is not clear and there is no systematic and comprehensive research on the compatible regularity of this prescription.Therefore,experiments to explore the compatible regularity of CSGS in the antidepressive effect would be designed to make a thorough research based on metabolomics via chronic unpredictable mild stress model of rat in depression.METHODS:1.Healthy 80 male SD rats were randomly divided into 8 groups: normal control group,depression model group,fluoxetine group,CSGS group,CSGS eliminated monarch group(NJ),CSGS eliminated minister group(NC),CSGS eliminated assistant group(NZ)and CSGS eliminated guide group(NS).Besides the normal group,the rats in the other 7 groups were established chronic stress depression model by chronic unpredictable mild stimulation(CUMS)for continuous 32 days.2.The body weight,open-field test and sugar preference test were measured to evaluate the change of rats behavioral.3.Elisa tests were used to investigate the effect of CUMS on serum5-hydroxytryptamine(5-HT),norepinephrine(NE),adrenocorticotrophin(ACTH)and corticosterone(CORT)level.4.To find the difference of serum metabolites between the model group and the normal control group,and analyze the possible relationship between the potential biomarkers and the depression syndrome,our study were based on the metabolomics by ultra-performance liquid chromatography quadrupole/time of flight mass spectrometry(UPLC-Q/TOF MS)and combined with statistical analysis and online metabolome database.Furthermore,we could characterize the changes of endogenous metabolites in the serum of rats after treatment with CSGS and its compatibility,in order to find the potential biomarker and target which may be related to the anti-depression effect.RESULTS:1.On the 32 nd day of CUMS,compared with control group,the body weight of rats in other 7 groups were decreased significantly(p<0.05 or p<0.01).Body weight from high to low ranked as follows: control>CSGS>fluoxetine>NS>NJ>NZ>NC>model.Compared with control group,the time of horizontal movement and vertical movement in OFT of rats in other 7 groups were reduced obviously(p<0.01).The time of horizontal movement and vertical movement in OFT from high to low ranked as follows: control>fluoxetine>CSGS>NS>NJ>NC>NZ>model.On the 31 st day of CUMS,compared with control group,the sugar preference in SPT of rats in other 7 groups were reduced remarkably(p<0.05 or p<0.01).The sugar preference in SPT from high to low ranked as follows: control>fluoxetine>CSGS>NS>NJ> NC>NZ> model.2.On the 32 nd day of CUMS,compared with control group,the 5-HT level of rats serum in other 7 groups were fell dramatically(p<0.01).The 5-HT level from high to low ranked as follows: control>fluoxetine>CSGS>NS>NJ>NC>NZ>model.Compared with control group,the NE level of rats serum in other 7 groups were fell significantly(p<0.05 or p<0.01).The NE level from high to low ranked as follows: control group>fluoxetine group>CSGS group>NS group>NC group>NZ group>NJ group>model group.Compared with control group,the ACTH level of rats serum in other 7 groups were raised markedly(p<0.05 or p<0.01).The ACTH level from high to low ranked as follows: model>NZ>NC>NJ>NS>CSGS>fluoxetine>control.Compared with control group,the CORT level of rats serum in other 7 groups were raised obviously(p<0.05 or p<0.01).The CORT level from high to low ranked as follows: model>NZ>NC> NS>NJ>CSGS>fluoxetine>control.3.Partial least squares discriminant analysis(PLS-DA)showed that the metabolic profile of the rat serum in model group was deviated from the control group,which suggested that CUMS could induced the change in the metabolic profile of rats.A total of 24 metabolites were characterized as potential biomarkers involved in CUMS-induced depression,which were corresponding to the dysfunctions of five pathways including glycerophospholipid metabolism,sphingolipid metabolism,arachidonic acid metabolism,selenoamino acid metabolism and fatty acid metabolism.4.Among them,16 may correlate to the regulation of CSGS treatment in depression,which were related to 16 proteins and 63 genes that may be the important target of CSGS treatment in CUMS-induced depression.According to the potential biomarker that were related to the overall antidepressant effect of CSGS,monarch drug involved 5 potential biomarkers,minister drugs involved 5potential biomarkers,assistant drug involved 6 potential biomarkers,guide drug involved 2 potential biomarkers.CONCLUSION:1.In this study,the chronic unpredictable mild stress rat model was estabilished successfully,which can simulate the symptoms of clinical depression.2.CSGS improved the abnormal index of depression,and the antidepressant effect of CSGS was considerable with fluoxetine.Also,the antidepressant effect of CSGS was better than that of its compatibility.3.The monoamine neurotransmitters and the nerve endocrine in serum of rats were disordered by CUMS modeling.The antidepressant effect of CSGS and its compatibility were regulated by the secretion of monoamine neurotransmitters(5-HT,NE)and hormones of HPA axis(ACTH,CORT).4.The profile of serum metabolism in CUMS model rats was changed significantly.The metabolomic pathways of rat,including sphingolipid metabolism,arachidonic acid metabolism,glycerophospholipid metabolism,selenoamino acid metabolism and fatty acid metabolism were disturbed obviously by CUMS model,which could be recovered after CSGS treatment.5.Lack of each functional unit(monarch,minister,assistant and guide)in CSGS had different degree on anti-depressant effect via different metabolomic pathways.The maximum influence on the regulation of metabolic profile in CUMS model rats was NZ group,and the minimal influence on that was NS group. |
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