Glycerol as a precursor for hepatic de novo glutathione synthesis in human liver

Background: Glycerol is a substrate for gluconeogenesis and fatty acid esterification in the liver, processes which are upregulated in obesity and may contribute to excess fat accumulation. Glycine and glutamate, in addition to cysteine, are components of glutathione, the major antioxidant in the liver. In principle, glycerol could be incorporated into glutathione via the TCA cycle or 3-phosphoglycerate, but it is unknown whether glycerol contributes to hepatic de novo glutathione biosynthesis. Methods: Glycerol metabolism to hepatic metabolic products including glutathione was examined in the liver from adolescents undergoing bariatric surgery. Participants received oral [U–¹³C₃]glycerol (50 mg/kg) prior to surgery and liver tissue (0.2–0.7g) was obtained during surgery. Glutathione, amino acids, and other water-soluble metabolites were extracted from the liver tissue and isotopomers were quantified with nuclear magnetic resonance spectroscopy. Results: Data were collected from 8 participants (2 male, 6 female; age 17.1 years [range 14–19]; BMI 47.4 kg/m² [range 41.3–63.3]). The concentrations of free glutamate, cysteine, and glycine were similar among participants, and so were the fractions of ¹³C-labeled glutamate and glycine derived from [U–¹³C₃]glycerol. The signals from all component amino acids of glutathione - glutamate, cysteine and glycine - were strong and analyzed to obtain the relative concentrations of the antioxidant in the liver. The signals from glutathione containing [¹³C₂]glycine or [¹³C₂]glutamate derived from the [U–¹³C₃]glycerol drink were readily detected, and ¹³C-labelling patterns in the moieties were consistent with the patterns in corresponding free amino acids from the de novo glutathione synthesis pathway. The newly synthesized glutathione with [U–¹³C₃]glycerol trended to be lower in obese adolescents with liver pathology. Conclusions: This is the first report of glycerol incorporation into glutathione through glycine or glutamate metabolism in human liver. This could represent a compensatory mechanism to increase glutathione in the setting of excess glycerol delivery to the liver.