List of metabolites from rat myocardium tissues in both MTX treated and untreated conditions
Cardiotoxicity is a well-established adverse effect of several drugs across multiple therapeutic indications. It is particularly prevalent following anticancer therapy. In order to evaluate the changes in metabolism associated with cardiotoxicity, we treated Wistar rats with a single high dose of methotrexate, and after five days, the animals were sacrificed. We then analyzed the cardiotoxicity parameters in serum and cardiac tissue with the goal of identifying a metabolic signature of cardiotoxicity using discovery-based metabolomics. The results showed a total of 95 metabolites that were found to be significantly (p < 0.05) modulated: either up- or downregulated in the HDMTX-treated group when compared with the control group. Based on these metabolites, it was found via integrated pathway analysis that a majority of the metabolites were associated with many important cardiac tissue metabolic pathways, such as the malate aspartate shuttle, taurine and hypotaurine metabolism, betaine metabolism, spermidine and spermine biosynthesis, and homocysteine degradation. Among them, L-arginine, homocysteine, and betaine were significantly upregulated, suggesting their role in cardiac tissue injury. The metabolic signature of toxicity may offer a novel approach to the prediction of elevated betaine levels as the novel biomarker for cardiotoxic potential due to HDMTX cardiac toxicity.
Steps to reproduce
Subject Health and medical sciences Specific subject area Cancer Research Type of data Table How data were acquired mass spectrometry (GCMS), Instruments: Agilent 7600, Masshunter, XCMS, MetaboAnalystR version 3.0.. Make and model and of the instruments used: Data format Analyzed Data Parameters for data collection Derivatization of myocardium extracts: The stored, dried, extracted samples of the upper layer of the heart tissue were warmed at room temperature (25°C) for 10 minutes prior to derivatization. The samples were derivatized with 25 μL of methoxyamine hydrochloride in pyridine (Sigma Aldrich) (20 mg/ml) at 37°C for 90 min with agitation (40rpm) in a water bath shaker. The second step of derivatization was performed by adding 40 μL of N methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) (Sigma-Aldrich) and incubating at 37°C for 30 min with agitation (40 rpm) in the water bath shaker. Samples were subjected to GC/MS analysis directly after derivatization. GC-MS analysis condition: Using a 10 μL syringe, 2 μL of each sample was injected into the split/splitless injector manually. The injection programs included washing the syringe before and after sample injection, the removal of air bubbles by sample pumping and an air buffer for the removal of the sample from the syringe after injection. The capillary column properties were 35% phenyl-coated silica, a column with a length of 30 meters and a thickness of 0.32 mm I.D. and 0.25 μm. Description of data collection GC-MS operation procedure: The GC oven was heated at 10°C/min from 60°C to 325°C, with 1 min being the initial time and 10 min being the final time. It ran for 37.5 min and then was cooled down to 60°C. The ion source heat was adjusted to 220°C. The energy of the electron was 70 eV. Splitless and split conditions were used for sample injection. Helium was used as the carrier gas flushed out at a flow of 10.5 ml/min for 1 min, with a saver run for 3 min at a 20 ml/min rate. A mass selective detector (MSD) was used at a 20 Hz signal data rate and set at 290°C for the transfer line. MS was operated on after 5.90 min of solvent delay time. Data source location Institution: King Abdulaziz University City/Town/Region:Jeddah Country:Saudi Arabia