ABSTRACT
AIM: The kynurenine (KYN) pathway is implicated in diseases such as cancer, psychiatric, neurodegenerative and autoimmune disorders. Measurement of KYN metabolite levels will help elucidating the involvement of the KYN pathway in the disease pathology and inform drug development. METHODOLOGY: Samples of plasma, cerebrospinal fluid or brain tissue were spiked with deuterated internal standards, processed and analyzed by LC-MS/MS; analytes were chromatographically separated by gradient elution on a C18 reversed phase analytical column without derivatization. CONCLUSION: We established an LC-MS/MS method to measure 11 molecules, namely tryptophan, KYN, 3-OH-KYN, 3-OH-anthranilic acid, quinolinic acid, picolinic acid, kynurenic acid, xanthurenic acid, serotonin, dopamine and neopterin within 5.5 min, with sufficient sensitivity to quantify these molecules in small sample volumes of plasma, cerebrospinal fluid and brain tissue.
Subject(s)
Brain/metabolism , Kynurenine/blood , Kynurenine/cerebrospinal fluid , Neopterin/blood , Neopterin/cerebrospinal fluid , Tryptophan/blood , Tryptophan/cerebrospinal fluid , Animals , Chromatography, High Pressure Liquid/methods , Humans , Kynurenine/analogs & derivatives , Kynurenine/metabolism , Mice, Inbred C57BL , Neopterin/metabolism , Quinolinic Acid/blood , Quinolinic Acid/cerebrospinal fluid , Quinolinic Acid/metabolism , Signal Transduction , Tandem Mass Spectrometry/methods , Tryptophan/metabolism , ortho-Aminobenzoates/blood , ortho-Aminobenzoates/cerebrospinal fluid , ortho-Aminobenzoates/metabolismABSTRACT
BACKGROUND: The mechanism of action of vagus nerve stimulation (VNS) in intractable epilepsy is not entirely clarified. It is believed that VNS causes alterations in cytokines, which can lead to rebalancing the release of neurotoxic and neuroprotective tryptophan metabolites. We aimed to evaluate VNS effects on tryptophan metabolites and on epileptic seizures and investigated whether the antiepileptic effectiveness correlated with changes in tryptophan metabolism. METHODS: Forty-one children with intractable epilepsy were included in a randomized, active-controlled, double-blind study. After a baseline period of 12 weeks, all children underwent implantation of a vagus nerve stimulator and entered a blinded active-controlled phase of 20 weeks. Half of the children received high-output (therapeutic) stimulation (n=21), while the other half received low-output (active control) stimulation (n=20). Subsequently, all children received high-output stimulation for another 19 weeks (add-on phase). Tryptophan metabolites were assessed in plasma and cerebrospinal fluid (CSF) by use of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and compared between high- and low-output groups and between the end of both study phases and baseline. Seizure frequency was recorded using seizure diaries. Mood was assessed using Profile of Mood States (POMS) questionnaires. RESULTS: Regarding tryptophan metabolites, anthranilic acid (AA) levels were significantly higher at the end of the add-on phase compared with baseline (p=0.002) and correlated significantly with improvement of mood (τ=-0.39, p=0.037) and seizure frequency reduction (τ=-0.33, p<0.01). No significant changes were found between high- and low-output groups regarding seizure frequency. CONCLUSION: Vagus nerve stimulation induces a consistent increase in AA, a neuroprotective and anticonvulsant tryptophan metabolite. Moreover, increased AA levels are associated with improvement in mood and reduction of seizure frequency.
Subject(s)
Epilepsy/metabolism , Epilepsy/therapy , Tryptophan/metabolism , Vagus Nerve Stimulation/methods , Adolescent , Affect , Biotransformation , Child , Child, Preschool , Double-Blind Method , Drug Resistance , Electrodes, Implanted , Female , Humans , Kynurenine/metabolism , Male , Metabolic Networks and Pathways , Seizures/epidemiology , Seizures/prevention & control , Treatment Outcome , Tryptophan/blood , Tryptophan/cerebrospinal fluid , ortho-Aminobenzoates/cerebrospinal fluid , ortho-Aminobenzoates/metabolismABSTRACT
Anthranilic acid (AA) has been attracted considerable attention as one of the L-tryptophan-kynurenine pathway metabolites in the central nervous system. In this study, the concentration of L-kynurenine (L-KYN) and AA in serum and CSF, and its related enzyme activities were determined in several species. In rabbits, CSF AA concentrations were lower and serum AA concentrations were slightly higher than those in other species. However, the concentrations of L-KYN were substantially higher in rabbits in both serum and CSF compared with other species. Tissue enzyme activities varied among species. In rabbits, lung IDO activities were higher, but liver kynurenine 3-hydroxylase activities were lower than those of the other species tested. Furthermore, brain kynurenine 3-hydroxylase activities were higher in gerbils than those in other species. These results clearly demonstrated that kynurenine pathway enzyme activities and metabolite concentrations vary with species.
