Biochemical Effects Of Venlafaxine On Astrocytes As Revealed By ~1HNMR-based Metabolic Profiling

BackgroundMajor depressive disorder(MDD)is a common and disabling illness affecting an increasing percentage of the world's population.MDD has a high prevalence,presents a significant threat to global public health,and has been shown to worsen when comorbid with other chronic diseases(e.g.,diabetes,hypertension,and arthritis).Adequate treatment and prevention of depression has significant implications.The diagnosis of major depressive disorder are clinical symptoms of patients,what's more,the underlying pharmacological mechanisms remain largely unclear.Venlafaxine,which exhibits a strong serotonin(5-HT),norepinephrine and weaker dopaminergic reuptake inhibition effect,has been successfully used for clinical treatment of MDD and resistance to depression as a representative of a new class of antidepressants(serotonin-norepinephrine reuptake inhibitors [SNRIs]).However,the cellular and molecular mechanism underlying rapid antidepressant actions are likely to be complex and have not been characterized to date.Metabonomics,an approach that enables thousands of metabolites in biological samples to be quantified simultaneously,has been extensively used to capture drug specific metabolic alterations in order to explore pharmacological mechanisms.It's significant to implement the novel and well-established metabonomics approach to determine the pharmacological mechanisms of venlafaxine.Ingenuity Pathway Analysis(IPA)is a web-based pathway analysis tool that employs advanced pathway enrichment analysis to confirm the most relevant metabolic pathways in cellular processes and diseases.Astrocytes are major component cells of the central nervous system(CNS)and are morphologically and functionally diverse.To date,numerous findings have reported the central function of astrocytes in the CNS.Astrocyte pathology in MDD has a close relationship with energy metabolites and the function of neurotransmitters.In this study,we investigated whether venlafaxine mainly targeted astrocytes to exert its therapeutic effects.A ~1HNMR-based metabonomics method,coupled with IPA analysis,was used to characterize the cellular metabolic profile of astrocytes treated with venlafaxine in vitro.Our primary objective was to identify metabolic perturbations in venlafaxine-induced cells to help us better understand the underlying molecular mechanisms of depression therapy with venlafaxine.ObjectiveVenlafaxine has been widely used in the clinical diagnosis of depression as a new antidepressant drugs,emerging evidence suggests that astrocytes have close relationship with the antidepressant effect of venlafaxine,while its underlying pharmacological mechanisms remained largely unclear.To investigate the metabolic pathways of fluoxetine is a significant way to explore the mechanism of fluoxetine to major depressive disorder.Methods:1.We have established an efficient culture method for purifying astrocytes.2.We selected the most suitable concentration of venlafaxine by the CCK-8 test which is a cell proliferation assay.(the suitable concentration is 10?M).3.We used a high-resolution proton nuclear magnetic resonance(~1HNMR)with different statistical analysis such as: principal component analysis(PCA),partial least squares-discriminate analysis(PLS-DA)and then analysis by IPA to explore the inner change of astrocytes which may provide some novel research ideas for the treatment of depression.Results1.Successful build efficient astrocyte culture method.2.After the multivariate statistical analysis,there are 31 metabolites has been found which have obvious difference.Venlafaxine can cause the increases of lactate,lysine,isobutyrate,myo-inositol,sebacate,tyrosine,unsaturated lipid,mannose-6-phophate,3-phosphoglycerate and glycerol-3-phopshate(G-3-P),and lower levels of ?-glucose,?-glucose,asparagine,ethanolamine,methionine,methylamine,ethanol,betaine,cholate,creatine,glutamate,glutamine,phosphocholine,glycoprotein,low density lipoprotein(LDL),very low-density lipoprotein(VLDL),phosphorylethanolamine,phytate,and fructose-6-phosphate.These metabolites have close relationship with energy metabolism,lipid metabolism and amino acid metabolism.3.We can know the different metabolic pathway and metabolites between venlafaxine and fluoxetine according the IPA analysis.The main metabolic pathway of both them is amino acid metabolism,and methionine,lysine and tyrosine are the core metabolites.ConclusionIn conclusion,explore the metabolic rule of venlafaxine to astrocytes by 1HNMR metabonomics to find out some different metabolites and use the IPA analysis show that energy metabolism,lipid metabolism and amino acid metabolism are the most important.This research may provide a further idea about the mechanism of antidepressant drugs.