| Rationales:Depression is one of prevalently serious mental disorders caused by various factors.And the most severely depressive illness can lead to suicide,which severely disrupts us in terms of work and life.Because of the complex mechanism and various causes of depression,the treatment of depression mainly relies on synthetic antidepressants based on Neurotransmitter Theory nowadays,but they are not satisfying with a variety of side effect,slow onset and expensive price.Xiaoyaosan is one of the best-known Traditional Chinese used for treatment for depression.However,as Chinese Medicine Prescription,the mechanism of Xiaoyaosan for antidepression is little known and there is no objective evaluation system for it in depression management.As an important branch of system biology,metbolomics can timely and truly represents the overall response of organism under pathological conditions and the stimulation of various drugs,which has obvious advantages in the overall efficacy evaluation and mechanism study of Chinese herbal medicine.In our previous study,the blood and urine of CUMS rats by pre-or post-treatment of Xiaoyaosan for 28 days was investigated by metabolomics based on 1H NMR,GC-MS andLC-MS and achieved some progress.But depression is especially complex in clinic,so human has some differences from animal models even the one widely recognized.This brings some difficulties for clinical study of metabolomics.So,in our previous study,a clinical study on antidepressant of Xiaoyaosan was investigated from two points of view including western medicine and traditional Chinese medicine on basis of previous animal experiments.And the plasma and urine of all subjects were also investigated by metabolomics based on 1H NMR,where some metabolites and corresponding pathways associated with antidepressant effect of Xiaoyaosan were found.These works play a very important role in facilitating the therapeutic evaluation and mechanism study of Xiaoyaosan.However,the study based on GC-MS and LC-MS metabolomics of plasma in all subjects has no done.Objective:To screen the plasma metaboliteschanged significantly for therapeutic response in depressed patients by Xiaoyaosan by GC-MS and LC-MS metabolomic analysis and then excavate the corresponding pathways associated with therapeutic response of Xiaoyaosan to reveal the mechanism of Xiaoyaosan for depression management;To screen the potential biomarkers with great clinical diagnostic value and associated with antidepressant effect of Xiaoyaosan in combination with those found previously in rats and depressed patients,which will facilitate to explain the mechanism of Xiaoyaosan and establishment of objective index for the evaluation of antidepressant effect of XYS.Methods:In this study,the plasma of healthy control was used as reference.The plasma of depressed patients before and after Xiaoyaosan administration was depressed patients were analyzed by metabolomics based on GC-MS and LC-MS.The data obtained from GC-MS and LC-MS analysis was introduced into SIMCA-P 13.0 software package.And then the metabolites changed significantly were screened based on variable importance in the projection(VIP)of S-plot and t test.At the same time,their pathways were uncovered by database called KEGG and published literatures.The clinical diagnostic values of these metabolites changed significantly were investigated by ROC analysis.So,the therapeutic evaluation and mechanism study of depression were done by metabolomics based on GC-MS and LC-MS.Finally,the potential biomarkers were determined via GC-MS and LC-MS after comparing with those found in rats and depressed patients previously,which will be facilitating to mechanism explaining and efficacy of Xiaoyaosan for depression management.Results:1.The GC-MS metabolism analysis method of clinical plasma was established.And plasma of all subjects was analyzed by GC-MS metabolomics coupled with multivariate statistical analysis.It was shown that the plasma levels of Oxalic acid and stearic acid of depressed patients increased significantly(p?0.05)compared healthy controls,while the levels of valine and urea decreased significantly(p?0.05).And the levels of the four metabolites were reversed to normal after administration of Xiaoyaosan,which is involved in glyoxylate and dicarboxylate metabolism,arginine and proline metabolism,fatty acid biosynthesis,valine,leucine and isoleucine biosynthesis,indicating that Xiaoyaosan played an important role in regulation of these biomarkers and their corresponding pathways to make an antidepressant management.These metabolites changed significantly were also evaluated their diagnostic ability based on the receiver operating characteristic(ROC)analysis.The AUCs for oxalic acid and stearic acid were>0.7,indicating a great clinical diagnostic value.The combination of the two metabolites changed significantly in discovery data set and prediction date set were 0.889 and 0.893,respectively.It was shown that the combination of the biomarkers can provide higher predictive power than single ones.2.The LC-MS metabolism analysis method of clinical plasma was established.And plasma of all subjects was analyzed by LC-MS metabolomics coupled with multivariate statistical analysis.20 metabolites changed significantly and associated with antidepressant of Xiaoyaosan,including LPCs(LPC(C10:3),LPC(C16:1),LPC(21:4),LPC(C19:0),LPC(C18:0)),acylcarnitines(carnitine(C14:2),carnitine(C10:4)),sphingomyelin,ceramides(stearic amide and palmitic amide),cystathionine,4-oxohex-2-enedioic acid and acetylphosphate,were found.It was involved in glycerophospholipid metabolism,fatty acid metabolism,arginine and proline metabolism,fatty acid metabolism,taurine and hypotaurine metabolism and cysteine and methionine metabolism.The AUCs for 4-oxohex-2-enedioic acid,creatinine,acetyl phosphate and cystathionine were>0.7,indicating a great clinical diagnostic value.The combination of the two metabolites changed significantly in discovery data set and prediction date set were 0.879 and 0.833,respectively.It was shown that the combination of the biomarkers can provide higher predictive power than single ones.3.In this study,30 potential biomarkers related to theclinically effective antidepressant of Xiaoyaosan were identified,including amino acids(alanine,valine,leucine),organic acids(lactate and oxalic acid),amine(trimethylamineoxide,glutamine and ceramides),fatty acid(stearic acid,heptadecadienoic acid),lipids(LPCs,sphingolipids,choline),acyl carnitines(carnitine C10:4 and carnitine C14:2),glucose and so on.These potential biomarkers were mainly involved in metabolism of three aspects including:energy metabolism(glyoxylate and dicarboxylate metabolism,pyruvate metabolism and fatty acid metabolism),neurotransmitter synthesis(valine,leucine and isoleucine biosynthesisglutamate and glutamine metabolism,arginine and proline metabolism,cysteine and methionine metabolism,taurine and hypotaurine metabolism,gycerophospholipid metabolism,sphingolipid metabolism and gut microbe metabolism.The ROC curve analysis showed that the AUCs of 8 plasmatic metabolites were more than 0.7 including plasmatic lactate,trimethylamine oxide,oxalic acid,stearic acid,4-oxohex-2-enedioic acidand creatinine,acetylphosphate and cystathionine creatinine,acetyl phosphate,indicating a great clinical diagnostic value.It will play an important role in the establishment of evaluating therapeutic effect of Xiaoyaosan.Conclusion:the therapeutic effect of Xiaoyaosan in depressive disorder is mainly related to energy metabolism,neurotransmitter synthesis and gut microbe metabolism.The AUCs of lactate,trimethylamine oxide,oxalic acid,stearic acid,4-oxohex-2-enedioic acid and creatinine,acetylphosphate and cystathionine in plasma of depressed patients were more than 0.7,indicating a great clinical diagnostic value.Metabolomics shows a great potential in mechanism explaining and efficacy evaluation of Xiaoyaosan for depression management. |
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