The identification of lipid metabolites was performed by comprehensively matching the extracted mass spectral features (including accurate precursor mass and MS/MS fragmentation patterns) against extensive online databases, predominantly mzCloud, HMDB, and LipidMAPS, as well as an in-house local spectral library. The identification confidence was secured by evaluating mass error tolerances and fragmentation matching scores. Downstream pathway analysis and flux visualization were subsequently executed using a custom Python script to map the lipid dynamic changes accurately.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative - reverse-phase","Liquid Chromatography MS - positive - reverse-phase"],"chromatography_protocol":["1. Untargeted Lipidomics (Isotope Tracing): Chromatographic separation was performed on a Waters ACQUITY UPLC CSH C18 column (2.1 mm _ 100 mm, 1.7 μm) maintained at 55°C. The mobile phase consisted of (A) acetonitrile/water (60:40, v/v) and (B) isopropanol/acetonitrile (90:10, v/v), both containing 10 mM ammonium formate and 0.1% formic acid. The flow rate was 0.26 mL/min with an injection volume of 3 μL. The gradient was: 0-1.5 min, 32% B; 1.5-4.0 min, 32%-45% B; 4.0-6.0 min, 45%-58% B; 6.0-12.0 min, 58% B; 12.0-16.0 min, 58%-70% B; 16.0-17.0 min, 70%-85% B; 17.0-20.0 min, 85% B; 20.0-26.0 min, 85%-95% B; 26.0-28.0 min, 95%-32% B; and 28.0-30.0 min, 32% B.
2. Targeted Acylcarnitine Profiling: Chromatographic separation was achieved using a Shimadzu LC-40D X3 UHPLC system on a Waters ACQUITY UPLC CSH C18 column (2.1 mm _ 100 mm, 1.7 μm) maintained at 50°C. The mobile phase consisted of (A) 0.1% formic acid in water and (B) 0.1% formic acid in acetonitrile. The flow rate was 0.25 mL/min with an injection volume of 5 μL. The gradient elution was: 0 min, 5% B; 0-11 min, 5%-100% B; 11-12 min, 100%-5% B; and 12-15 min, 5% B.
"],"publication":["Tracer-based Metabolomics Technology to Investigate the Role and Mechanism of Fangji Huangqi Tang in Regulating Lipid Metabolism Disorders in MPC-5 Cells."],"submitter_affiliation":["Shanxi university"],"submitter_name":["zhaowei xue"],"organism_part":["cell line"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["The metabolite extraction was performed using an optimized chloroform/methanol system. Briefly, 400 μL of ice-cold methanol and 800 μL of chloroform were added to the harvested cell pellet, followed by the addition of 240 μL of LC-MS grade water. The mixture was vortexed thoroughly and then centrifuged at 12,000 rpm for 10 minutes at 4°C to induce phase separation. The lower organic phase, which primarily contains nonpolar and neutral lipids, was carefully collected. To ensure exhaustive recovery of lipid species, the remaining precipitate was re-extracted with 200 μL of cold methanol, vortexed for 60 seconds, and centrifuged again. The combined extracts were dried under vacuum. Finally, the residue was reconstituted in 140 μL of dichloromethane/methanol (2:1, v/v). After vortexing for 1 minute and ultrasonication at 4°C for 10 minutes, the samples were subjected to a final centrifugation (12,000 rpm, 10 min, 4°C), and the clear supernatant was transferred to glass vials for LC-MS analysis.
"],"organism":["Mus musculus"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS14277"],"author":["Xuemei Qin. Modern Research Center for Traditional Chinese Medicine of Shanxi University. Modern Research Center for Traditional Chinese Medicine of Shanxi University, Taiyuan 030006, P.R. China.. qinxm@sxu.edu.cn.","Aiping Li. Modern Research Center for Traditional Chinese Medicine of Shanxi University. Modern Research Center for Traditional Chinese Medicine of Shanxi University, Taiyuan 030006, P.R. China.. aipingli@sxu.edu.cn.","zhaowei xue. xuezhaowei123@163.com."],"data_transformation_protocol":["1. Untargeted Lipidomics (Isotope Tracing): Raw LC-MS data were processed using Compound Discoverer 3.0 (Thermo Fisher Scientific). The 'Stable Isotope Labeling with Metabolika Pathways and ID' workflow was used for feature extraction and alignment. Natural isotope abundance was corrected to calculate the Relative Exchange ratio (e.g., M16 (%) = [M16 / (M0 + M1 + ... + M16)] _ 100%).
2. Targeted Acylcarnitine Profiling: Raw data acquired from the SCIEX Triple Quad 5500 in MRM mode were processed for peak integration and targeted absolute quantification. Acylcarnitine concentrations were calculated based on the calibration curves established using representative carnitine standards.
"],"study_factor":["Treatment"],"submitter_email":["xuezhaowei123@163.com"],"sample_collection_protocol":["The murine podocyte cell line MPC-5 was cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% Penicillin-Streptomycin at 37°C in a humidified atmosphere of 5% CO2. For the construction of the in vitro injury model, cells were treated with 0.10 μg/mL Adriamycin (ADR) for 24 hours. For the stable isotope labeling experiment, the optimized tracer 13C16-palmitic acid (conjugated with fatty acid-free BSA) was added to the culture medium at a final concentration of 75 μM. To investigate the therapeutic effects, the model cells were concurrently co-incubated with different doses of Fangji Huangqi Tang (FHT) extract (0.45 mg/mL or 0.60 mg/mL), or positive control drugs including Dexamethasone/Atorvastatin (0.50 μM / 1.0 μM) and Triptolide/Atorvastatin (1.0 ng/mL / 1.0 μM). All groups were incubated for 24 hours. Following incubation, the cells were washed twice with ice-cold PBS, harvested, flash-frozen, and stored at -80°C for subsequent lipidomic analysis.
"],"omics_type":["Metabolomics"],"study_design":["Metabolomics","Mus musculus","Waters ACQUITY UPLC H-Class System","targeted analysis","untargeted analysis","Shanxi University","Lipid Metabolism","Compound Discoverer","nephrosis","Thermo Scientific Q Exactive HF","cell line","nephrotic syndrome"],"curator_keywords":["Metabolomics","Mus musculus","Waters ACQUITY UPLC H-Class System","targeted analysis","untargeted analysis","Shanxi University","Lipid Metabolism","Compound Discoverer","nephrosis","Thermo Scientific Q Exactive HF","cell line","nephrotic syndrome"],"mass_spectrometry_protocol":["1. Untargeted Lipidomics (Isotope Tracing): Mass spectrometric data were acquired using a Q Exactive high-resolution mass spectrometer (Thermo Fisher Scientific) equipped with an electrospray ionization (ESI) source. The instrument was operated in a positive/negative switching mode using a Full MS/dd-MS2 (Top N = 5) strategy. The scan range was set to m/z 80-1200. The mass resolutions were set at 70,000 for Full MS and 17,500 for MS2. The stepped normalized collision energies (NCE) were 25, 35, and 45 eV. Other specific parameters were as follows: automatic gain control (AGC) target value of 1_10^5, capillary temperature of 300°C, sheath gas flow rate of 45 arb, and auxiliary gas flow rate of 10 arb. The spray voltages were maintained at 3,500 V for positive mode and 3,000 V for negative mode.
2. Targeted Acylcarnitine Profiling: Targeted mass spectrometric analysis was conducted using a SCIEX Triple Quad 5500 mass spectrometer equipped with an ESI source. Data acquisition was performed in multiple reaction monitoring (MRM) mode. The primary source parameters in positive ion mode were optimized as follows: IonSpray Voltage at 5500 V, TurboIonSpray Temperature (TEM) at 500°C, Curtain Gas (CUR) at 40 psi, Nebulizer Gas (GS1) at 50 psi, and Heater Gas (GS2) at 50 psi.
"],"additional_accession":[]},"is_claimable":false,"name":"Tracer-based Metabolomics Technology to Investigate the Role and Mechanism of Fangji Huangqi Tang in Regulating Lipid Metabolism Disorders in MPC-5 Cells","description":"Lipid metabolism disorders play a critical role in the progression of nephrotic syndrome (NS). Although the traditional Chinese medicine formula Fangji Huangqi Tang (FHT) has been shown to restore lipid homeostasis in vivo, its specific molecular targets and effects on dynamic lipid metabolism in podocytes remain unclear. To address this, we developed a 13C16-palmitic acid stable isotope tracer lipidomics platform combined with targeted acylcarnitine metabolomics and molecular biology approaches to investigate the regulatory mechanisms of FHT against Adriamycin-induced lipotoxicity in MPC-5 podocytes. Our results demonstrate that FHT protects podocytes by remodeling lipid metabolism__romoting phospholipid synthesis while inhibiting triglyceride accumulation, and modulating fatty acid metabolism through enhanced mitochondrial β-oxidation and suppressed de novo lipogenesis. These changes reversed mitochondrial dysfunction and preserved podocyte structural integrity.","dates":{"publication":"2026-09-20","submission":"2026-04-14"},"accession":"MTBLS14277","cross_references":{}}