A high-throughput liquid chromatography-tandem mass spectrometry assay for the simultaneous quantification of p -cresol sulfate, p -cresol glucuronide, indoxyl sulfate, and indoxyl glucuronide in HepaRG culture medium and the demonstration of mefenamic acid as a potent and selective detoxifying agent.
| Autor: | Al-Dajani AR; Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Alberta, Canada., Kiang TKL; Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Alberta, Canada. |
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| Jazyk: | angličtina |
| Zdroj: | Expert opinion on drug metabolism & toxicology [Expert Opin Drug Metab Toxicol] 2024 Sep 29, pp. 1-13. Date of Electronic Publication: 2024 Sep 29. |
| DOI: | 10.1080/17425255.2024.2409257 |
| Abstrakt: | Background: p -cresol and indole are uremic compounds which undergo sulfonation to generate the highly toxic p -cresol sulfate ( p CS) and indoxyl sulfate (IxS). They are also subjected to glucuronidation to produce the less toxic p -cresol glucuronide ( p CG) and indoxyl glucuronide (IG). We developed and validated an assay to quantify these metabolites in HepaRG cells. We also tested the effects of mefenamic acid on their in-situ formations in relation to the development of cellular necrosis. Research Design and Methods: HepaRG cells were exposed to p -cresol or indole (0-1 mM) with mefenamic acid (0-3000 nM) for 24 hours to generate uremic metabolites. Cells were also exposed to 0.5 mM p -cresol or indole with/without 30 nM mefenamic acid to characterize lactate dehydrogenase (LDH) release. Results: The assay exhibited high sensitivity and wide calibration ranges covering human concentrations. HepaRG cells also generated physiologically-relevant concentrations of each metabolite. Mefenamic acid inhibited p CS formation in a concentration-dependent manner without affecting p CG, IxS, or IG. Mefenamic acid also reduced LDH release from p -cresol (by 50.12±5.86%) or indole (56.26±3.58%). Conclusions: This novel assay is capable of quantifying these metabolites in HepaRG cells. Our novel findings suggest that mefenamic acid can be potentially utilized therapeutically to attenuate p CS-associated toxicities. |
| Databáze: | MEDLINE |
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