MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14868/m_MTBLS14868_LC-MS_alternating_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14868/a_MTBLS14868_LC-MS_alternating_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14868/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14868/s_MTBLS14868.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14868<p>Multiple reaction monitoring (MRM) has been used for quantification of screening fragment ions.</p>MetaboLightsPublicLiquid Chromatography MS - alternating - reverse-phase<p>The LC/MS portion of the platform was based on a Shimadzu Nexera X2 LC-30AD system equipped with an ACQUITY UPLC HSS T3 column (1.7 μm, 2.1 mm×100 mm, Wasters) and a triple quadruple mass spectrometer (5500 QTRAP, AB SCIEX). Metabolites were detected in electrospray negative-ionization and positive-ionization mode. The 2 μL samples were injected sequentially with LC autosampler. The ACQUITY UPLC HSS T3 column (1.7 μm, 2.1 mm×100 mm, Wasters) was heated to 40 °C under a flow rate of 200 μL/min. A gradient was used to separate the compounds consisted of 0.1% Formic acid aqueous solution (solvent A) and 100% acetonitrile (solvent B). The gradient started at 100% solvent A for 2.5 min and increasing linearly to 70% solvent A over 9 min, and then increasing linearly to 0% solvent A over 1 min following with a 5.4 min hold before returning the starting mixture during 0.1min and re-equilibrating for 2.5 min. QC samples were injected every several samples during acquisition to monitor the reproducibility of the test.</p>targeted metabolomics of rat's serum.The First Affiliated Hospital, and College of Clinical Medicine of Henan University of Science and TechnologyXuejiao Liblood serummass spectrometry assay<p>The metabolites were extracted from serum with 1 mL precooled methanol/acetonitrile/water (v/v, 2:2:1) under sonication for 1 h in ice baths. The mixture was incubated at -20 °C for 1 h followed by centrifugation at 16,000 g, 4 °C for 20 min, and then transferred to the sampling vial for LC-MS analysis.</p>Rattus norvegicushttps://www.ebi.ac.uk/metabolights/MTBLS14868Xuejiao Li. The First Affiliated Hospital, and College of Clinical Medicine of Henan University of Science and Technology. white_snow1987@163.com.Chuanxin Liu. The First Affiliated Hospital, and College of Clinical Medicine of Henan University of Science and Technology. 9906194@haust.edu.cn.<p>The discriminating metabolites were obtained using a statistically significant threshold of variable influence on projection (VIP) values obtained from the OPLS-DA model and two-tailed Student’s t test (p value) on peak area of each metabolite for two groups comparison. The p value was calculated by one-way analysis of variance (ANOVA) for multiple groups analysis. Metabolites with VIP&gt; 1 and p value&lt; 0.05 were considered to be statistically significant. The significantly differential metabolites were used to perform cluster analyses with R package.</p>TreatmentDosewhite_snow1987@163.com<p>The origin of samples is serum. Stored in -80 degree.</p>MetabolomicspodophyllotoxinMetabolomicsThermo Scientific Q Exactivetargeted analysisRattus norvegicusblood serumSerumShimadzu Nexera UHPLC systemexperimental blanktargeted metabolite profilingpodophyllotoxinMetabolomicsThermo Scientific Q Exactivetargeted analysisRattus norvegicusblood serumSerumShimadzu Nexera UHPLC systemexperimental blanktargeted metabolite profiling<p>Widely targeted metabolites were quantified using multiple-reaction monitoring mode set up as previous report described (Cao,&nbsp;G.,&nbsp;et&nbsp;al. 2020; Zheng,&nbsp;F.,&nbsp;et&nbsp;al. 2020). All transitions from large-scale metabolites (MT1000 Kit developed by Shanghai BioProfile) were detected with optimized decluttering potential and collision energy. The MS conditions were set as follows: negative-ionization mode: Source Temperature 550℃, Ion Source Gas1 (GAS1): 40, Ion Source Gas2 (GAS2): 50, Curtain Gas (CUR): 35, Ion Spray Voltage Floating (ISVF) :-4500V; positive-ionization mode: Source Temperature 550℃, Ion Source Gas1 (GAS1): 40, Ion Source Gas2 (GAS2): 50, Curtain Gas (CUR): 35, Ion Spray Voltage Floating (ISVF) :5500V.</p>DEOXYCARNITINEdGDPbeta-Hydroxymyristic acidAminohippuric acidPG(18:0/18:1)4-HydroxynonenalDL-2-Aminooctanoic acidCorticosteronePhenylacetylglutamineLPE(18:2)RiboflavinDocosatetraenoic acidFolic acidButyric acidPC(16:0/18:2)EctoineKetoleucineCEAFlavoneD-glucosamine 1-phosphateOxoglutaric acidCarnitine-C8DCLPE(18:3)BiotinCitrullineCELalpha-Muricholic acid1-MethylxanthineD-Glucuronic acidOctadecanedioic acidHyocholic acid3-Hydroxyanthranilic acidHexanoylcarnitineDhCer d18:0/22:0Hexose PoolGlycinePC(20:0/18:1)EstradiolL-Pipecolic acidLPE(18:0)HordenineMevalonic acidArginine13S-hydroxyoctadecadienoic acidSM 42:0;2PI(18:0/20:4)D-Glucose 1-phosphate2'-Deoxycytidine diphosphateSM 32:2;3N6-acetyl lysineFA(18:0)LPE(18:1)4-Ethyl-2-methoxyphenol2-Ketoglutaric acidOleamide3-phosphoglycerateTiglylcarnitineDG(16:0/22:6)N-Acetyl-5-hydroxytryptamineGOLAN-Acetylneuraminic acidThiazolidine-4-carboxylic acid4-Pyridoxic acidPantothenic acidGlycerol 3-phosphateSM 34:1;2GuanosineCitrateGlyceraldehyde 3-phosphateFA(18:1)beta-D-GlucosamineXanthurenic acidD-Glucosamine 6-phosphateShikimic acidCer d18:1/18:1Carnitine-C10:1PA(16:0/18:1)Shikimate-3-phosphateCarnitine-C10:2Acetoacetic acidGlyceric acidcis-aconitic acidGamma-glutamyl-Alanine1alpha,25-Dihydroxyvitamin D3AlanylphenylalanineFA(18:2)Prostaglandin D2PyridineUridine 5-monophosphateUridineOrnithineD-Gluconic acidRetinalSM 40:2;3Vitamin B13-UreidopropanoateSM 34:0;2IsovalerylcarnitineSM 34:0;3IndoleSerineArgpyrimidineProlineFA(18:3)6-Ketolithocholic acid5-Hydroxy-6,8,11,14,17-eicosapentaenoic acidLactamide2-Hydroxybenzoic acidInosine diphosphateAdenosineL-ThyroxineL-Ascorbic acidPS(18:0/18:1)dIMPN-Acetyl-L-glutamic acidN-AcetyltryptophanUric acidHomogentisic acidPimelylcarnitine11,12-DiHETrE17-OH-ProgesteroneN-Acetyl-D-glucosamine 1-phosphatetetranor-Prostaglandin E MetabolitS-Carboxymethyl-L-cysteineHypotaurineLPC(18:0)6-Phospho-D-gluconateBenzoic acidMelatoninAla-LeuMannoseGlucoseQuinolinic Acidgamma-GlutamyltyrosineGlycylproline2-Pyrocatechuic acidCortexoloneCMASpermidinePS(20:0/18:1)FA(14:0)SphingosineD-Glyceraldehyde 3-phosphateN-Acetyl-L-tyrosineCarnitine-C16:1gamma-GlutamylleucineTaurocholic acidCML2-Isopropylmalic acidThreonineTaurodeoxycholic acid Pool2-Ketobutyric acidIsodeoxycholic acid5-AminolevulinateCL(18:2/18:2/18:1/18:1)LPE(16:1)Carnitine-C4DCUrocanic acid11-Dehydro-thromboxane B2Valeric acidAbietic acidLysineL-CystathionineMethylsuccinic AcidN-Fructosyl tyrosineHydroxyprolineL-AbrineSM 34:2;3Carnitine-C125'-MethylthioadenosineCarnitine-C104-NitrophenolPI(18:1/20:5)Carnitine-C16Carnitine-C14Alpha-ketoisovaleric acid1,5-anhydroglucitolCarnitine-C18PC(18:2/16:1)CreatinineTaurolithocholic acidXanthosineAspartateN-Carbamoyl-L-aspartatePyridoxine3-Methyl-2-oxovaleric acidUridine 5'-monophosphateN-Fructosyl phenylalanineHexacosanoic acid5-Aminopentanoic acid3-HydroxyhexanoylcarnitineValineSuberic Acid2-ThiocytidinePA(16:0/18:2)P-Toluenesulfonic acidHexaethylene glycolHydroxyisocaproic acidGlutathione disulfideN6-lactoylPhosphotyrosineTaurochenodeoxycholic AcidGlycocholic acid5-MethyltetrahydrofolateItaconic acidTauro-alpha/beta/omega muricholic acidCarnitine-C10 OHFlavin adenine dinucleotide3-methoxytyramineDeoxyribose phosphateCarnitine-C14:2PG(18:1/16:1)AMPPC(18:1/18:1)Cinnamoylglycine2-Hydroxy-3-methylbutyric acidGlutamateAsp-PheN6-formylN-Acetyl-L-aspartic acidDG(18:1/18:1)n-Caproic acid8-hydroxyguanosineN-methylphenethylaminePyroglutamyl-IsoleucineSM 36:0;23alpha-Hydroxy-12 Ketolithocholic Acid sulfatePI(16:0/18:0)23-Nordeoxycholic acidCL(18:2/18:2/18:2/16:1)Lipoxin?A41-MethylhistidinePC(18:0/18:1)Glyoxylic acidGamma-glutamyl-CysteinePE(P-18:0/16:0)FA(16:0)SphinganineSM 42:2;2Taurolithocholic acid 3-sulfateSM 42:2;37-Ketolithocholic acidIndolelactateThymineGlycochenodeoxycholic acidFA(16:2)OleoylethanolamideCarnitine-C3DCDodecylbenzenesulfonic acid1-Methyladenosine7-methylguanineFA(16:1)4-Hydroxyphenylacetic acidSarcosineSM 36:1;2ProgesteronePE(18:0/22:5)TaurineAllantoin3,4-Dihydroxyphenylacetic acidGlucaric acidO-PhosphoethanolamineLithocholic acidCholic acid sulfateL-HomoserineL-Threonine4-Guanidinobutyric acidPhosphoenolpyruvateKynurenic acidN-FormylmethionineAgmatineGamma-glutamyl-LeucineFA(22:2)PalmitoylethanolamidePyrralineCarnitine-C0MethionineEstriol(E3)N-Acetyl-D-glucosaminePC(16:1/18:1)Oxaloacetic acidPE(P-18:1/18:1)Carnitine-C5OHPE(14:0/18:2)SM 36:2;22-Naphthalenesulfonic acidcAMPUDP-glucuronateCystineN-AcetylglutamineLPE(22:6)Dehydrolithocholic acidCAFFEIC ACIDFA(12:0)Lithocholenic acid sulfatePentadecanoic acidMyo-InositolPhenylalanineSerotonin(5-L-Glutamyl)-L-glutamineIsoleucylisoleucinePhenylacetylglycineS-Nitroso-L-glutathionePC(16:0/22:6)PE(P-18:1/18:3)Ferulic acidEpiandrosteroneCholestenoneLeucylprolineDG(18:2/20:5)gamma-Glutamyl-L-methionineEicosapentaenoic acidAconitateCarnitine-C18:28,9-Epoxyeicosatrienoic acidL-KynurenineCreatine7-methylguanosineSphingosine 1-phosphateCarnitine-C8:1SM 38:0;2PhytosphingosineOctulose-monophosphateN6-glycerinyl3-Methyluridine3-Hydroxykynurenine2-AcetylpyrazinePC(14:0/18:2)Glycodeoxycholic acid sulfateCarnitine-C18:1D-Glucose 6-phosphatebeta-Muricholic acidPE(18:2/18:2)FA(22:6)HomoarginineFA(24:0)2'-Deoxyuridine 5'-triphosphateN6-oxalylN2,N2-Dimethylguanosine12-Ketolithocholic acidDG(14:0/18:3)Quinolin-2,8-diol11-Hydroxyeicosatetraenoic acid2-keto-D-Gluconic acidLPE(22:5)3-Indolepropionic acidSepiapterinGABASuccinic acidHypoxanthineN6-methyladenosine8(R)-hydroxy- eicosatetraenoic acidPhenyllactic acidL-MalateAdipic acidAcetic acid2,4-DinitrophenolGluconolactonePE(18:2/18:3)PhenylalanylisoleucineGuanidinoacetic acid3-FORMYLINDOLEFA(24:3)Hippuric acidDesmosterolN-Acetylaspartylglutamate(S)-4,5-DihydroorotateSedoheptulose monophosphateN6-threonylD-Ala-D-alaSM 38:1;2N,N-DimethylanilineSM 38:1;3Homovanillic acidCarnitine-C2Carnitine-C4Carnitine-C3Carnitine-C6Carnitine-C5Carnitine-C8PE(P-18:0/22:6)CatechinPyruvateFructoseNicotinuric acidCarnitine-C4OHHeptadecanoic acidInosineNorepinephrinePurineDimethylglycineTaurocholic acid-3-O-sulfateSerylleucineGlutaminePE(18:1/18:1)N-Acetyl-L-phenylalanineFA(20:1)Dihydroxyacetone phosphateCholic acid11,12-Epoxyeicosatrienoic acidSM 38:2;3Histidine20-Hydroxyeicosatetraenoic acidAnserineUreidosuccinic acidSaccharopineNa-Methyl-L-ornithine monohydrochloride3-MethylhistidineDG(16:1/18:2)CaffeinePC(18:0/22:5)4-Hydroxybenzoylcholine(S)-12-Hydroxyeicosatetraenoic acidTyramineSM 40:0;23-HydroxybenzaldehydeDG(16:0/18:3)Allocholic acidFA(20:0)N-acetylserineVanillylmandelic acidFarnesyl diphosphate5-hydroxyeicosatatraeniocacidMalondialdehyde3, 5-TetradecadiencarnitineSebacic acid5,6-DHETbeta-Sitosterol3,4-dihydroxyphenylalanineN4-AcetylcytidineN-AcetylhistamineGlycerophosphocholineErgothioneineNG,NG-Dimethyl-L-arginineLeucyl-leucineS-Methyl-L-cysteineN-AcetylornithinePicolinic acidTarginineAspartylglycosamineBetaine aldehydeLPE(20:4)PyridoxamineSalicylic acidAlanineL-Argininosuccinic acidFA(20:2)TryptophanXanthineDeoxyguanosineD-GlucosamineQuinic acidCholineS-D-LactoylglutathioneAsparagineN-Acetyl-L-alanine3-Aminoisobutyric acid2-Ketohexanoic acidDiphenylamineAldosteroneSM 30:2;4D-Fructose 6-phosphateOrotic acidTiglylglycineL-Dihydroorotic acid4-HydroxyphenyllactateChenodeoxyglycocholic acidNicotinamide N-oxideLTB4LPE(20:2)UDPPE(18:2/16:1)7-MethylxanthineIsobutyrylglycineInositolLPI(18:2)Hexose Monophosphate PoolFA(20:4)Indoxyl sulfateIndole-3-acetaldehydePE(18:0/18:1)N1-Methyl-2-pyridone-5-carboxamideGlycyl-L-leucineS-AdenosylhomocysteineGamma-glutamyl-IsoleucineIsocitrateCer d18:1/26:05'-DeoxyadenosineLPE(20:3)LPG(18:1)2-PiperidoneIsovalerylglycineFA(22:1)2-MethylcitrateDhCer d18:0/24:1Epinephrine (E)Allantoic acidThromboxane B22,3-Diphospho-D-glyceratep-CresolDihydrothymineAzelaic acidCarnitine-C5:13-MethylcytidineRiboseHydroxybenzoic acidp-Aminobenzoic acidL-2-Aminoadipic acidSalicyluric acidgamma-L-Glutamyl-L-phenylalanineImidazoleacetic acidFA(22:0)5-HydroxyindoleacetateGALADeoxycholic acid2,3-Dihydroxybenzoic acidCarnitine-C18:1OHIndole-3-carboxylic acidThreonateDeoxycytidineCarbamoyl phosphate14,15-Epoxy-5,8,11-eicosatrienoic acidGlycodeoxycholic acid5-Hydroxy-L-tryptophan15-Hydroxyeicosatetraenoic acidBiopterinHomocystinefalsetargeted metabolomics of rat's serumPodophyllotoxin (PPT) exhibits limited clinical utility due to its nephrotoxicity, and its underlying mechanisms remain poorly understood. This study employs the toxicological evidence chain (TEC) framework and integrated multi-omics analyses to investigate the potential involvement of the microbiota-gut-kidney (MGK) axis in PPT- induced nephrotoxicity in SD rats. Toxicity was systematically evaluated through longitudinal monitoring of body weight, general behavior, biochemical markers, intestinal barrier function, and histopathological alterations. In parallel, multi-omics analyses, encompassing microbiome, metabolomics, and transcriptomics, were conducted to delineate the mechanistic underpinnings. The results showed that PPT exposure induced pronounced renal and intestinal damage, manifesting as significant weight loss, diarrhea, elevated renal injury biomarkers, increased lipopolysaccharide (LPS) levels, and diamine oxidase (DAO), along with histopathological lesions and enhanced apoptosis in renal and colonic tissues. PPT exposure perturbed gut microbiota homeostasis, characterized by depletion of beneficial taxa (e.g., Lactobacillus) and enrichment of potentially pathogenic genera (e.g., Bacteroides and Aggregatibacter), concomitant with diminished short-chain fatty acid (SCFA) production and altered metabolite profiles in fecal, serum, and renal samples. Integrated multi- omics analysis further revealed activation of the JAK1/2-STAT3 signaling pathway, upregulation of pro-inflammatory mediators (TNF-α, IL-6, IL-1β, LPS, and TMAO), and suppression of anti-inflammatory cytokines (IL-10 and IL-4). These in vivo molecular and inflammatory patterns were partially reproduced in HK-2 cells co-cultured with fecal microbiota supernatant from PPT-treated rats. In addition, the JAK1/2 inhibitor ruxolitinib attenuated PPT-induced JAK1/2-STAT3 phosphorylation and inflammatory cytokine secretion in HK-2 cells. Correlation network analysis further identified associations between gut dysbiosis, systemic inflammation, and metabolic perturbations. Collectively, these findings support a mechanistic hypothesis that MGK- axis disruption and JAK1/2-STAT3 signaling may contribute to PPT-associated nephrotoxicity. However, in vivo interventional studies are required to establish definitive causality.2026-06-272026-06-27MTBLS14868