Glutathione conjugates of 4-hydroxy-2(E)-nonenal as biomarkers of hepatic oxidative stress-induced lipid peroxidation in rats

摘要: Here we present a simple, specific, and sensitive liquid chromatography/mass spectrometry method to measure 4-hydroxy-2(E)-nonenal-glutathione (HNE-GSH), the major stable hepatic metabolite of HNE after GSH conjugation, as marker oxidative stress in rat liver hepatocytes. Commonly employed methods for measurement lipid peroxidation-derived free aldehydes or modified proteins suffer from artificial formation adducts cellular molecules during sample preparation derivatization, resulting an overestimation background levels. Basal levels HNE-GSH tissue untreated rats were detected amounts 20 pmol/g liver. Rats exposed single dose iron nitrilotriacetate (Fe(III)NTA; 15 mg Fe/kg bw, ip), model compound induction stress, revealed fivefold increase compared controls 5 h dosing. Moreover, significant HNE-mercapturic acid (HNE-MA) its reduced DHN-MA was evident at 24 treatment, which also reflected increased plasma concentrations these secondary metabolites. In agreement with vivo data, time-dependent <1 123 +/- 16 pmol/10(6) cells over hepatocytes treated Fe(III)NTA (150 microM). An <1.0 7.2 0.3 could be observed allyl alcohol (500 microM, 3 h), known generation These data suggest that direct conjugation product primary metabolites may represent reliable biomarker stress-induced peroxidation vivo.