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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/m_MTBLS14272_LC-MS_positive_reverse-phase_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/m_MTBLS14272_LC-MS_negative_reverse-phase_metabolite_profiling-1_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/m_MTBLS14272_LC-MS_positive_hilic_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/m_MTBLS14272_LC-MS_negative_reverse-phase_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/a_MTBLS14272_LC-MS_negative_reverse-phase_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/a_MTBLS14272_LC-MS_positive_hilic_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/a_MTBLS14272_LC-MS_positive_reverse-phase_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/s_MTBLS14272.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272/a_MTBLS14272_LC-MS_negative_reverse-phase_metabolite_profiling-1.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14272Thermo Scientific Xcalibur Workstation version 4.0 (Thermo Fisher Scientific, Rockford, IL, U.S.A.) and Compound Discover version 3.3 software (Thermo Fisher Scientific, Rockford, IL, U S.A.) were used to process mass spectrometry data. Importing raw data into Compound Discoverer for peak alignment, retention time correction, peak area extraction, and structural identification of metabolites.MetaboLightsPublicLiquid Chromatography MS - negative - reverse phaseLiquid Chromatography MS - positive - HILICLiquid Chromatography MS - positive - reverse phaseSeparations were conducted utilizing the Vanquish UHPLC system(Thermo Scientific, USA). ACQUITYTM Premier CSH Phenyl-Hexyl Column (1.7 μm, 2.1*100 mm, Waters, Milford, MA, U.S.A.) and AtlantisTM Premier BEH Z-HILIC Column (2.5 μm, 2.1*100 mm, Waters, Milford, MA, U.S.A.) were used for chromatographic separations respectively. Two different mobile phase systems were used on the Phenyl-Hexyl Column analysis: system 1 were 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B), system 2 were 5 mM ammonium acetate in water (A) and acetonitrile (B), and the same gradient elution conditions for both systems were set as follows: 0-2 min, 5%B-5%B; 2-13 min, 5%B-95%B; 13-15 min, 95%B-95%B; 15-15.1 min, 95%B-5%B; 15.1-18 min, 5%B-5%B. The mobile phases during Z-HILIC Column analysis were 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B), and the gradient elution conditions were set as follows: 0-2 min, 95%B-95%B; 2-10 min, 95%B-60%B; 10-12 min, 60%B-95%B; 12-15 min, 95%B-95%B. Throughout the analysis, the column temperature was set at 40 °C, the flow rate was 0.3 mL/min, and the injection volume was 2 μL. Pentose phosphate pathway-derived NADPH facilitates physiological hypertrophy and alleviates ischemia/reperfusion injury in the heart.No. 99 Shangda Road, Baoshan District, ShanghaiYufan ChaoCardiac Tissuemass spectrometry assayThe extraction of heart tissue samples is as follows: Firstly, the frozen heart tissues were ground a powder using a cryogenic grinder. Next, add 100 μL of extraction solvent (acetonitrile: methanol: water = 2:2:1, containing 1μg/mL Internal standard) kept on ice per 10 mg of ground tissue. The mixture was vortexed, and centrifuged twice at 13,000 rpm for 10 minutes at 4°C. Finally, the second supernatant was collected and transferred to an MS vial for analysis. Additionally, 15 μL of each sample was aspirated and pooled to create a quality control (QC) sample.Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS14272Qingxun Hu. School of Medicine, Shanghai University. 99 Shangda Road, Shanghai, 200444, China. qingxh@shu.edu.cn.Thermo Scientific Xcalibur Workstation version 4.0 (Thermo Fisher Scientific, Rockford, IL, U.S.A.) and Compound Discover version 3.3 software (Thermo Fisher Scientific, Rockford, IL, U S.A.) were used to process mass spectrometry data. Importing raw data into Compound Discoverer for peak alignment, retention time correction, peak area extraction, and structural identification of metabolites. Exercisefychaosw@163.comMice swam in tanks with ~32oC and 15 cm depth water. In brief, mice swam in a ramp protocol starting at 10 min twice a day, with 10 min increase each day until 90 min twice a day was reached. The protocol ended after 4 weeks. Twenty-four hours after the final swimming session, the exercised mice were euthanized, and their tissues were collected for further analysis.MetabolomicsThermo Scientific Vanquish UHPLC SystemMetabolomicsMus musculusThermo Scientific Q Exactiveuntargeted analysishigh-performance liquid chromatography-mass spectrometryCardiac Tissueexperimental blankPentose Phosphate PathwayThermo Scientific Vanquish UHPLC SystemMetabolomicsMus musculusThermo Scientific Q Exactiveuntargeted analysishigh-performance liquid chromatography-mass spectrometryCardiac Tissueexperimental blankPentose Phosphate PathwayMass spectrometry analysis was conducted using a Q-Exactive Plus Orbitrap MS mass spectrometer (Thermo Scientific, USA) equipped with a heated electrospray ionization source (HESI). Mass spectrums were obtained with a full scan with a resolution of 70,000, the mass range of 70-1500 m/z and an AGC target value of 3e6. For the QC sample, the parallel reaction monitoring (PRM) and ddMS2 data were acquired at a resolution of 17,500, with AGC target values of 2e5 and 1e5, respectively. Mass spectrometry was detected in both positive and negative ion modes. The settings for the ion source parameters were as follows: spray voltage of 3.8 kV for positive and 3.2 kV for negative; sheath gas and auxiliary gas composed of nitrogen (purity ≥ 99.99 %) with flow rates of 30 arb and 10 arb, respectively; capillary temperature of 320 °C; and auxiliary gas heater temperature of 300 °C.falseMetabolomic data related to the alleviation of cardiac ischemia/reperfusion injury by NADPH derived from pentose phosphate pathwayExercise promotes physiological cardiomyocyte growth and protects against ischemia/reperfusion (I/R) injury in the heart. The molecular mechanism by which exercise benefits cardiac metabolism and function remains largely unknown. Exercise (swimming or treadmill)-induced physiological hypertrophy models were established in mice. Pentose phosphate pathway (PPP) flux was analyzed by 1,2-13C2 glucose stable isotope tracing. In vitro and in vivo manipulations of glucose-6-phosphate dehydrogenase (G6PD) and NADPH were conducted by using lentivirus- and adeno-associated virus-mediated gene transfer, respectively. Myocardial I/R injury model was established in adult mice. Genetically encoded fluorescent indicators (iNap, roGFP, and Hyper) were used to monitor compartmentalized redox and reactive oxygen species (ROS) in neonatal cardiomyocytes during oxygen glucose deprivation/reperfusion (OGD/R). An iNap biosensor-based high-throughput screen was conducted to identify new agents that elevate NADPH levels in cardiomyocytes.2026-05-142026-04-14MTBLS14272