APPLICATION OF TARGETED LIPIDOMICS TO DETERMINE CHANGES IN THE PLASMA LIPIDOME OF MALE RATS FOLLOWING REPEAT ORAL ADMINISTRATION OF METHAPYRILENE

Význam tématu

Targeted lipidomics provides sensitive profiling of lipid alterations that occur in response to pharmacological intervention.
It enables detection of molecular perturbations relevant for drug safety assessment and biomarker discovery.

Cíle a přehled studie

This study evaluated changes in the plasma lipidome of male Wistar rats following repeat oral administration of methapyrilene at 0, 50, and 150 mg/kg/day over a five-day period.
Dose-response effects were monitored at 24, 72, and 120 hours post-dosing.

Použitá metodika a instrumentace

Sample collection via vena cava at D1, D3, D5 and pooling for QC.
Protein precipitation using IPA/ACN with stable isotope-labeled standard mix (Avanti EQUISPLASH).
8-minute HILIC UPLC separation (Waters ACQUITY Premier UPLC with Premier BEH Amide Column).
Quantification of 430 lipid species via targeted MRM in positive and negative ion modes on Waters Xevo TQ-Absolute tandem quadrupole MS.
Data processing in Skyline and statistical analysis in MetaboAnalyst 6.0.

Hlavní výsledky a diskuse

PCA revealed no separation at D1 and D3 for lower dose but clear group separation at D5 for both 50 and 150 mg/kg.
VIP plots highlighted ceramides, glucosyl ceramides, and carnitines as the most dysregulated lipids in positive mode; free fatty acids, bile acids, PI, PE, and LPEs in negative mode.
Significant upregulation of glycochenodeoxycholic acid (GCDCA), glycoursodeoxycholic acid (GUDCA), and glycodeoxycholic acid at high dose on D5.
Elevation of Hexadecanoyl-L-carnitine and Oleoyl-L-carnitine, and differential changes in free fatty acids (e.g., C20:0 decrease).
Method exhibited robust performance with QC CVs between 1.5% and 12%.

Přínosy a praktické využití metody

Rapid high-throughput workflow suitable for large-scale lipid biomarker screening.
Comprehensive coverage of diverse lipid classes enables mechanistic insights into drug-induced lipid perturbations.
Applicable to preclinical safety studies and translational research in toxicology and pharmacology.

Budoucí trendy a možnosti využití

Integration with other omics platforms (e.g., proteomics, metabolomics) for multi-omic biomarker discovery.
Expansion to clinical studies and personalized medicine applications.
Advances in data analysis using machine learning to extract deeper biological insights.

Závěr

The targeted HILIC-MRM method demonstrated rapid, reproducible, and sensitive detection of plasma lipid alterations following methapyrilene exposure.
Distinct lipid profile changes correlated with dosing and time, supporting its utility in safety assessment workflows.

Reference

1. Graichen ME, Neptun DA, Dent JG, Popp JA, Leonard TB. Effects of methapyrilene on rat hepatic xenobiotic metabolizing enzymes and liver morphology. Fundam Appl Toxicol. 1985;5:165–174.
2. Munjoma N, Isaac G, Muazzam A, et al. High Throughput LC-MS Platform for Large Scale Screening of Bioactive Polar Lipids in Human Plasma and Serum. J Proteome Res. 2022;21(11):2596-2608. doi:10.1021/acs.jproteome.2c00297.
3. Adams KJ, Pratt B, Bose N, et al. Skyline for Small Molecules: A Unifying Software Package for Quantitative Metabolomics. J Proteome Res. 2020;19(4):1447-1458. doi:10.1021/acs.jproteome.9b00640.
4. Pang Z, Lu Y, Zhou G, et al. MetaboAnalyst 6.0: towards a unified platform for metabolomics data processing, analysis and interpretation. Nucleic Acids Res. 2024;1-9. doi:10.1093/nar/gkae253.