MetaboLights

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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14295/m_MTBLS14295_LC-MS_positive_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14295/m_MTBLS14295_LC-MS_negative_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14295/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14295/a_MTBLS14295_LC-MS_negative_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14295/s_MTBLS14295.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14295/a_MTBLS14295_LC-MS_positive_reverse-phase.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14295The online KEGG, HMDB database was used to annotate the metabolites by matching the exact molecular mass data (m/z) of samples with those from database. If a mass difference between observed and the database value was less than 10 ppm, the metabolite would be annotated and the molecular formula of metabolites would further be identified and validated by the isotopic distribution measurements. We also used a in-house fragment spectrum library of metabolites to validate the metabolite identidification.Statistical analysis was performed in R (version 4.0.0). The raw protein intensity will be normalized by method 'medium', Hierarchical clustering was performed using pheatmap package. Principal component analysis (PCA) was performed using metaX package. The PLSDA analysis is performed by the R package ropls and the VIP values of each variable are calculated.Correlation analysis was performed by Pearson correlation coefficient of cor package .The three conditions of P Value<0.05, difference multiple >1.2 obtained by T test and VIP calculated by PLSDA analysis simultaneously met the screening of the final metabolites with significant differences..Hypergeometric-based enrichment analysis with KEGG Pathway was performed to annotate protein sequences. individually.The software GSEA (v4.1.0) and MSigDB were used for gene set enrichment analysis to determine whether a set of genes in a specific KEGG pathway in different situations. Meeting this condition |NES|>1, NOM p-val<0.05, FDR q-val<0.25 were considered to be significantly different between the two groups. .The network map is drawn according to the pathway where the metabolite is located.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseThe chromatographic separation was performed using an ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 µ m, Waters); Mobile phase A is 5 mmol/L ammonium acetate+5 mmol/L acetic acid aqueous solution, and mobile phase B is acetonitrile, The gradient elution procedure is: 0-1 min, 1% B, 1~9 min , 1% ~ 99% B, 9~9.1 min, 99% ~ 1% B; 9.1~12 min, 1% B, Flow rate of 0.35 mL/min; Injection volume 2 μ L; The chromatographic column temperature is 40 ℃.Semaglutide targets muscle mitochondria to regulate glutamine metabolism and treat osteoarthritis.Shanghai Sixth People's HospitalYuchen Tianplasmamass spectrometry assayThaw the sample on ice, take 100 μ L and add 400 μ L of ice MeOH/ACN (v: v, 1:1), mix well, precipitate the protein at -20 ° C for 1 hour, centrifuge at 13000rpm for 15 minutes at 4 ° C, take the supernatant and concentrate at 4 ° C, then dissolve again for testing. At the same time, take an equal volume of each sample and mix it to form a QC sample.Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS14295Yuchen Tian. Shanghai Sixth People's Hospital. yctian97s@sjtu.edu.cn.The acquired MS data pretreatments including peak picking, peak grouping, retention time correction, second peak grouping, and annotation of isotopes and adducts was performed using XCMS software. LC−MS raw data files were converted into mzXML format and then processed by the XCMS, CAMERA and metaX toolbox implemented with the R software. Each ion was identified by combining retention time (RT) and m/z data. Intensities of each peaks were recordedand a three dimensional matrix containing arbitrarily assigned peak indices (retention time-m/z pairs), sample names (observations) and ion intensity information (variables) was generated.C-mitoOAS-mitoyctian97s@sjtu.edu.cnExtract whole blood into an anticoagulant tube, centrifuge at 3000g for 15 minutes, and collect the supernatant plasmaMetabolomicsMetabolomicsMus musculusuntargeted analysisosteoarthritisVanquish Flex UHPLCTripleTOF 6600SemaglutideMuscleexperimental blankplasmaMetabolomicsMus musculusuntargeted analysisosteoarthritisTripleTOF 6600Vanquish Flex UHPLCSemaglutideMuscleexperimental blankplasmaUse the electric spray ion source to collect once each in positive and negative electric spray mode. The temperature of the ion source is 350 ℃; The capillary voltage is+3.8 kV in positive mode and -3.4 kV in negative mode; the purge gas flow rate is 0 Arb; The auxiliary airflow speed is 15 Arb; The sheath gas flow rate is 50 Arb. The data acquisition mode is full scan and data dependent acquisition (DDA). In one acquisition cycle, the full scan range is 70-1050 Da, the resolution is 70000, the automatic gain control (AGC) is 3E6, and the maximum ion implantation time (Maximum IT) is 100 ms. In the full scan, the first 5 ions with a response intensity higher than 100000 are selected for secondary scanning, with a resolution of 17500 and a maximum ion implantation time (Maximum IT) of 50 ms. The dynamic exclusion time is set to 6 seconds.falseSemaglutide targets muscle mitochondria to regulate glutamine metabolism and treat osteoarthritisMetabolic and inflammatory diseases often coexist, and become important challenges facing global health, and obesity has shown to be a susceptibility factor for osteoarthritis (OA). As a metabolic regulatory drug for lowering blood sugar and body weight, semaglutide has shown additional therapeutic effects in OA, but the underlying mechanism requires further investigation. Here we employed cross-tissue single-cell RNA sequencing (scRNA-seq) analysis, and found that semaglutide significantly improved mitochondrial metabolic disorders in muscle tissue under high-fat diet (HFD) and OA conditions in mice. The results of metabolome and mitochondrial proteomics indicated that semaglutide targeted muscle mitochondria to regulate glutamine metabolism during OA. Intramuscular injection of semaglutide pre-stimulated mitochondria from myoblasts could significantly alleviate OA inflammation and pain symptoms, which was achieved by inhibiting muscle glutaminase activity, upregulating circulating glutamine concentration, and thereby alleviating cartilage inflammation. In vitro experiments further confirmed the alleviating effect of glutamine produced by myoblasts stimulated by semaglutide on chondrocyte inflammation. These findings reveal the mitochondrial regulatory mechanism in the muscle-OA axis, and provide a new perspective for the application of semaglutide in OA treatment.2026-05-062026-04-16MTBLS14295