MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14381/m_MTBLS14381_LC-MS_alternating_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14381/a_MTBLS14381_LC-MS_alternating_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14381/s_MTBLS14381.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14381/i_Investigation.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14381<p>A mixed standard solution of 100 central carbon metabolites at multiple concentrations was used to establish standard curves. The R² value of the quantitative formula for each standard curve was greater than 0.995. Then, based on the ratio of the mass spectrometry intensity of each metabolite to that of the internal standard in each sample, the injection concentration for each sample was calculated. Using the reconstitution volume and the initial sample amount, the content of each central carbon metabolite in the original sample was finally determined. A total of 83 central carbon metabolites were detected in the experimental samples.</p>MetaboLightsPublicLiquid Chromatography MS - alternating - reverse-phase<p>Separation was performed using a Shimadzu Nexera X2 LC-30AD high-performance liquid chromatography system. The column was a Kinetex F5 column. Mobile phase A was 10 mM ammonium acetate, and mobile phase B was pure acetonitrile. Samples were kept in the autosampler at 4 degrees, the column temperature was 30 degrees, the flow rate was 200 μL/min, and the injection volume was 5 μL. The LC gradient was as follows: 0 to 3 min, B maintained at 0 percent; 3 to10 min, B increased linearly from 0 percent to 95 percent; 10 to 12 min, B maintained at 95 percent; 12 to13 min, B decreased linearly from 95 percent to 0 percent; 13 to 14 min, B maintained at 0 percent.</p>β-hydroxybutyrate Restores Social Deficits by Suppressing HDAC9 in the ACC of Shank3B-deficient Mice.Fourth Military Medical UniversityErling Hublood serummass spectrometry assay<p>Samples were thawed at 4 degrees, vortexed to mix, and 100 μL of serum was taken. 400 μL of prechilled 100 percent methanol was added, vortexed to mix, and then sonicated in an ice bath for 20 min. The mixture was left to stand at under 20 degrees for 1 h, followed by centrifugation at 16,000 g at 4 degrees for 20 min. The entire supernatant was collected and dried in a high speed vacuum concentrator. For mass spectrometry analysis, the dried sample was reconstituted in 100 μL of prechilled 50 percent methanol in water, centrifuged at 20,000 g at 4 degrees for 15 min, and the supernatant was taken for MS injection analysis.</p>house mousehttps://www.ebi.ac.uk/metabolights/MTBLS14381Erling Hu. Fourth Military Medical University. No.169, Changle West Road, Xi’an, Shaanxi, 710032, China. erling20@fmmu.edu.cn.<p>Chromatographic peak areas and retention times were extracted using MultiQuant software. Retention times were calibrated using standards for metabolite identification.</p>Dieterling20@fmmu.edu.cn<p>Four-week-old <em>Shank3B</em> KO mice were fed either a ketogenic diet (KD) or a standard diet (SD) for four consecutive weeks. Subsequently, peripheral blood was collected, serum was isolated, and the serum samples were stored at under 80 degrees.</p>MetabolomicsMetabolomicsAB SCIEX QTRAP 6500+targeted analysisliquid chromatography-mass spectrometryketogenic diet(R)-3-hydroxybutyratehouse mouseblood serumShimadzu Nexera UHPLC systemtargeted metabolite profilingautismexperimental sampleMetabolomicsAB SCIEX QTRAP 6500+targeted analysisliquid chromatography-mass spectrometryketogenic diet(R)-3-hydroxybutyratehouse mouseblood serumShimadzu Nexera UHPLC systemtargeted metabolite profilingautismexperimental sample<p>Mass spectrometry analysis was performed using a QTRAP 6500+ mass spectrometer (AB SCIEX) in both positive and negative ion modes. The ESI source parameters were as follows:</p><p>Positive ion mode: Source Temperature 500 °C, Ion Source Gas 1 (GAS1): 55 psi, Ion Source Gas 2 (GAS2): 60 psi, Curtain Gas (CUR): 35 psi, Ion Spray Voltage Floating (ISVF): 5500 V.</p><p>Negative ion mode: Source Temperature 500 °C, Ion Source Gas 1 (GAS1): 55 psi, Ion Source Gas 2 (GAS2): 60 psi, Curtain Gas (CUR): 35 psi, Ion Spray Voltage Floating (ISVF): –4500 V.</p><p>Multiple reaction monitoring (MRM) mode was used to detect the analyte ion pairs.</p>GTPalpha-Ketoglutaratec-di-AMPDihydroxyacetone phosphateOxaloacetateO-PhosphoethanolamineGlucoseisoleucineFructose 1,6-bisphosphateThiamine pyrophosphateGDPglycinePhosphoenolpyruvateXylulose 5-phosphateRibulose-1,5-bisphosphateSuccinatecystine2-phosphoglycerate1,5-Anhydro-d-mannitoldTMP4-HydroxyprolineasparagineglutaminecAMPUDP-GlcNAcornithineNADHUDPglucoseGlucose 1-phosphatealanineNADPTrehalose 6-phosphoric acidtryptophan3-phosphoglycerolFMNtrans-AconitatedCMPFructose 6-phosphateFumarate3-phosphoglycerateADPglucoseUracilATPhistidineserinedUTPSuccinyl-CoALactateRibulose 5-phosphateGuanosineCitrateLiothyronineGlyceraldehyde 3-phosphateADPShikimic acidUTPItaconic acidarginineGlucose 6-phosphateAcetoacetic acidaspartate2,3-Diphosphoglyceric acidleucineCis-AconitateAMP3-Phenyllactic acidGlutamateIMPCytidineprolinelysineMalatemethionineIsocitrateUridineUMPtyrosinecitrullineRibose 5-phosphateNADPH5,10-Methylene-THFErythose-4-phosphoric acidMalonyl-CoATetrahydrofolic acidphenylalaninePyruvateAdenineSedoheptulose 7-phosphate6-phosphogluconateAcetyl-CoA2-Hydroxyglutaric aciddAMPNADthreonineSarcosineArgininosuccinic acidInosinedUMPvaline3-Hydroxybutyric acidfalseβ-Hydroxybutyrate Restores Social Deficits via Suppressing HDAC9 in the Anterior Cingulate Cortex of Shank3B-Deficient Mice<p>Autism spectrum disorder (ASD) is characterized by core deficits in social behavior, yet effective interventions remain limited. Ketogenic diet (KD) shows behavioral benefits in ASD, but the underlying mechanisms remain unclear. Here, using Shank3B knockout (KO) mice, we find that KD induces systemic ketosis with marked elevation of β-hydroxybutyrate (BHB). Oral BHB alone recapitulates KD's prosocial effects, restoring social interaction and neuronal activity in the anterior cingulate cortex (ACC). Mechanistically, we identify HDAC9 as a region- and cell type–specific epigenetic target upregulated in ACC neurons of Shank3B KO mice and suppressed by BHB. HDAC9 overexpression in ACC neurons induces social and synaptic deficits, while class IIa HDAC inhibition phenocopies BHB effects. BHB also restores dendritic complexity, excitatory transmission, and AMPA receptor expression. These findings uncover a metabolite-driven epigenetic mechanism linking ketogenic metabolism to circuit-level rescue of social behavior, highlighting HDAC9 as a therapeutically actionable target for ASD.</p>2026-07-082026-04-26MTBLS14381MTBLC32484MTBLC16761MTBLC15422MTBLC17621MTBLC17552MTBLC15996MTBLC16908CHEBI:32484CHEBI:16761CHEBI:15422CHEBI:17621CHEBI:17552CHEBI:15996CHEBI:16908