Among the optimized MRM transitions per analyte, the Q1/Q3 pairs that showed the
highest sensitivity and selectivity were selected as “quantifier” for quantitative monitoring. The additional
transitions acted as “qualifier” for the purpose of verifying the identity of the target analytes. Analyst (1.7.3, SCIEX) and Biotree Biobud (v2.0.4) were employed for MRM data acquisition.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative - reverse-phase"],"chromatography_protocol":["The UHPLC separation was carried out using Agilent 1290 UPLC, equipped with a ACQUITY UPLC BEH C18 Column(1.7 µm, 2.1 mm all_fetch_status all_status eb_eye_copy_status eb_eye_entry_counts eb_eye_fetch_status eb_eye_metabolights_complete.xml eb_eye_metabolights_compounds.copy eb_eye_metabolights_compounds.xml eb_eye_metabolights_studies.copy e_fetch_status europe_PMC_metabolights_studies.copy europe_PMC_metabolights_studies.xml head.xml studies.copy study.xml tail.xml thomsonreuters_metabolights_studies.copy thomsonreuters_metabolights_studies.xml 150 mm). The mobile phase A was 0.01% formic acid in water, and the mobile phase B was0.01% formic acid in acetonitrile. The column temperature was set at 50 °C. The autosampler temperature was set at 6 °C, and the injection volume was 10 μL.
"],"publication":["13(S)-HODE Generated by Rhizopus microsporus var. rhizopodiformis Aggravates MASLD via the Elane-PAR2-SREBP-1c Axis."],"submitter_affiliation":["Hainan medical university"],"submitter_name":["Daya Zhang"],"organism_part":["supernate"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["After the addition
of 450 μL of extraction solution (0.5 M ammonium acetate, pH 5.5, pre-chilled to 4°C, containing BHT in
water, and an isotopically-labeled internal standard mixture), the samples were vortexed for 30 seconds and
sonicated for 5 minutes in an ice-water bath. Incubate the sample with shaking at 4°C for 30 minutes. A 750
μL aliquot of the extraction reagent (n-hexane/ethyl acetate = 2/3) was then transferred to an Eppendorf tube.
After vortexing for 5 minutes, the sample was centrifuged (5 minutes at 3500 rpm and 4°C). The supernatant
was transferred to a new Eppendorf tube. The extraction and centrifugation process was repeated twice,
and the supernatant liquids from the two separations were pooled. Finally, the samples were evaporated
to dryness under a gentle stream of nitrogen and reconstituted in 75 μL of a 30% acetonitrile/water (v/v)
solution. The reconstituted solutions were vortexed for 30 seconds. The reconstituted solution was then
transferred to an EP tube equipped with a filter membrane. After centrifugation (15 minutes at 12,000 rpm
and 4°C), the clear supernatant was subjected to UHPLC-MS/MS analysis.
"],"organism":["Rhizopus microsporus var.rhizopodiformis"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS15663"],"author":["Daya Zhang. Hainan medical university. 875025464@qq.com."],"data_transformation_protocol":["The raw data were converted to the mzXML format using ProteoWizard
and processed with an in-house program.
"],"study_factor":["Group"],"submitter_email":["875025464@qq.com"],"sample_collection_protocol":["150 μL aliquot of each individual sample was precisely transferred to an Eppendorf tube.
"],"omics_type":["Metabolomics"],"study_design":["Metabolomics","ProteoWizard msconvert","AB SCIEX Triple Quad 6500+","targeted analysis","Rhizopus microsporus var.rhizopodiformis","Agilent 1290 Infinity II UHPLC","targeted metabolite profiling","supernate","13(S)-HODE","Rhizopus microsporus var. rhizopodiformis"],"curator_keywords":["Metabolomics","ProteoWizard msconvert","AB SCIEX Triple Quad 6500+","targeted analysis","Rhizopus microsporus var.rhizopodiformis","Agilent 1290 Infinity II UHPLC","supernate","targeted metabolite profiling","13(S)-HODE","Rhizopus microsporus var. rhizopodiformis"],"mass_spectrometry_protocol":["Triple Quad 6500+ Mass Spectrometer (SCIEX, USA), equipped with an electrospray ionization interface, was applied for assay development. Typical ion source parameters were: Ion Spray Voltage = -4500 V, Ion Source Gas 1=30 psi, Ion Source Gas 2=30 psi, Temperature = 500 °C, Curtain Gas= 40 psi. The MRM parameters for each of the targeted analytes were optimized using flow injection analysis, by injecting the standard solutions of the individual analytes, into the ESI source of the mass spectrometer. Several most sensitive transitions were used in the MRM scan mode to optimize the collision energy for eachQ1/Q3 pair.
"],"metabolite_name":["12S-HEPE","9S-HOTrE","19S-HETE","2,3-dinor TXB2","¡À11-HDoHE","¡À9(10)-EpOME","15S-HEPE","17R-HDoHE","¡À8-HDoHE","¡À11-HEPE","15-keto PGE2","20-HETE","12-oxo LTB4","13S-HOTrE","16S-HETE","¡À16,17-EpDPE","ARA","11¦Â-PGF2¦Á","1a,1b-dihomo PGF2¦Á","PGF3¦Á","tetranor-12S-HETE","LTE4","5S-HETE","6-keto PGF1¦Á","12-epi-6-trans-LTB4","DHA","¡À9(10)-DiHOME","¡À17(18)-EpETE","PGB2","AT-RvD1","7-HDoHE","8-iso PGF2¦Á","¡À5-iso PGF2¦Á-VI","13S-HOTrE(¦Ã)","20-COOH-AA","8S,15S-DiHETE","¡À11(12)-EET","8-iso-15-keto PGF2¦Â","¡À13-HDoHE","14,15-LTE4","5S-HETrE","6S-LXA4","¡À14,15-DiHETrE","19R-hydroxy PGE2","2,3-dinor-8-iso-PGF2¦Á","2,3-dinor-11¦Â-PGF2¦Á","¡À11,12-DiHETrE","9-Nitrooleic Acid","15-keto PGF1¦Á","13,14-dihydro PGF2¦Á","¡À12(13)-EpOME","20-hydroxy LTB4","PGA2","¡À14(15)-EpETE","10-Nitrooleic Acid","5S,15S-DiHETE","5S,6R-DiHETE","PGF2¦Á","¦¤17-6-keto PGF1¦Á","PGJ2","15-deoxy-¦¤12,14-PGD2","PGD2","15-oxoEDE","PGD1","PGD3","LXA5","1a,1b-dihomo PGE2","12S-HETE","5S-HEPE","15S-HETE","9R-HETE","14,15-LTC4","13S-HODE","TXB3","15S-HETrE","TXB2","TXB1","¡À8,9-DiHETrE","LTB4","LXB4","11¦Â-13,14-dihydro-15-keto PGF2¦Á","18-HEPE","¡À20-HDoHE","11-dehydro TXB2","DTA","14,15-LTD4","PGF1¦Á","15-oxoETE","15-deoxy-¦¤12,14-PGJ2","13,14-dihydro-15-keto PGF2¦Á","12S-HHTrE","LXA4","PD1","17S-HETE","¡À19(20)-DiHDPA","20-carboxy LTB4","8S-HETrE","¡À12(13)-DiHOME","¡À19,20-EpDPE","7-epi-MaR1","¡À5,6-DiHETrE","RvE1","¡À14(15)-EET","LTD4","11¦Â-PGE2","¡À18-HETE","13-OxoODE","PGE3","PGE2","11S-HETE","13,14-dihydro-15-keto PGD2","PDX","PGE1","9-OxoODE","5-oxoETE","19R-hydroxy PGF2¦Á","14S-HDoHE","4-HDoHE","RvD5","RvD4","RvD3","RvD2","2-Mar","RvD1","EPA","12-oxoETE","6-trans LTB4","LTC4","9S-HODE","¡À8(9)-EET"],"additional_accession":[]},"is_claimable":false,"name":"13(S)-HODE Generated by Rhizopus microsporus var. rhizopodiformis Aggravates MASLD via the Elane-PAR2-SREBP-1c Axis","description":"Abstract Objective:Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) poses a major public health challenge. Increasing data suggest a role of gut fungi in liver disease, yet their diversity and biological function in MASLD are still poorly understood. Methods:Internal transcribed spacer (ITS) sequencing was conducted on fecal samples from 107 individuals with MASLD and 120 matched healthy controls (HC). A high-fat diet (HFD)-induced MASLD mouse model was established, and multi-omics analyses, including liver transcriptomic sequencing, qPCR, WB, and immunofluorescence, were performed to elucidate the molecular mechanisms underlying key gut fungi associated with MASLD. Microbial-derived metabolites involved in MASLD were investigated through multi-omics analyses in vitro and in vivo. Results:MASLD patients showed significantly reduced fungal diversity and richness compared with HC. The gut mycobiota of MASLD patients exhibited a marked increase in the pathogenic genus Rhizopus, particularly Rhizopus microsporus var. rhizopodiformis and R. microsporus var. chinensis, which were positively associated with hepatic steatosis and body mass index (BMI). Fecal microbiota transplantation from MASLD patients into antibiotic-treated mice induced MASLD-like phenotypes, while amphotericin B treatment mitigated disease progression. Oral administration of R. microsporus var. rhizopodiformis in HFD-fed mice aggravated gut dysbiosis, disrupted intestinal barrier integrity, and activated hepatic SREBP-1c signaling, leading to upregulation of key lipogenic enzymes ACC1 and FASN. These changes were mediated through a pro-inflammatory cascade involving the hepatic LPS–TLR4–NF-κB axis and enhanced neutrophil degranulation. 13(S)-HODE, potentially identified as a key metabolite produced by R. microsporus var. rhizopodiformis, aggravated MASLD. 13(S)-HODE promotes neutrophil trafficking to the liver, where released elastase (Elane) directly cleaves and activates the master transcription factor SREBP-1c via a non-canonical PAR2 signaling pathway, leading to uncontrolled hepatic de novo lipogenesis. Conclusion:Our findings suggest that Rhizopus microsporus var. rhizopodiformis and its candidate metabolite 13(S)-HODE are associated with MASLD progression, potentially via the Elane-PAR2-SREBP-1c axis. These observations provide a preliminary basis for further investigating the role of gut fungi in MASLD pathogenesis.","dates":{"publication":"2026-09-14","submission":"2026-09-14"},"accession":"MTBLS15663","cross_references":{}}