A metabolomic perspective of griseofulvin-induced liver injury in mice

Abstract

Griseofulvin (GSF) causes hepatic porphyria in mice, which mimics the liver injury associated with erythropoietic protoporphyria (EPP) in humans. The current study investigated the biochemical basis of GSF-induced liver injury in mice using a metabolimic approach. GSF treatment in mice resulted in significant accumulations of protoporphyrin IX (PPIX), N-methyl PPIX, bile acids, and glutathione (GSH) in the liver. Metabolomic analysis also revealed bioactivation pathways of GSF that contributed to the formation of GSF-PPIX, GSF-GSH and GSF-proline adducts. GSF-PPIX is the precursor of N-methyl PPIX. A six-fold increase of N-methyl PPIX was observed in the liver of mice after GSF treatment. N-methyl PPIX strongly inhibits ferrochelatase, the enzyme that converts PPIX to heme, and leads to PPIX accumulation. Excessive PPIX in the liver results in bile duct blockage and disturbs bile acid homeostasis. The accumulation of GSH in the liver was likely due to Nrf2-mediated upregulation of GSH synthesis. In summary, this study provides the biochemical basis of GSF-induced liver injury that can be used to understand the pathophysiology of EPP-associated liver injury in humans.

Ferrochelatase is a therapeutic target for ocular neovascularization

“…Griseofulvin has been used widely to treat fungal infections and is taken orally, often for months or years (Petersen et al, 2014;Liu et al, 2015). An off-target side effect of this therapy is that griseofulvin causes the formation of NMPP, along with other alkylated porphyrins, primarily in the liver (Liu et al, 2015). NMPP in turn acts as an active-site inhibitor of FECH (Cole & Marks, 1984).…”

Chronic Treatment with Isoniazid Causes Protoporphyrin IX Accumulation in Mouse Liver

“…11 Briefly, 100 mg of liver samples were homogenized in 400 μl water. In 100 μl of liver homogenate, 200 μl methanol was added and then vortexed two times for 1 min.…”

Optimization of extraction and analytical protocol for mass spectrometry‐based metabolomics analysis of hepatotoxicity

“…The optimization protocol of liver metabolites extraction included four experiment stages (Table 1). Five extraction solvents were chosen, designed and compared for liver metabolite extraction based on previous studies (Huang et al, 2011;Liu et al, 2015;Masson, Alves, Ebbels, Nicholson, & Want, 2010). Liver samples (50 mg) were first homogenized in 200 μL ice-cold water followed by addition of 300 μL methanol (MeOH; M1), 500 μL 50% acetonitrile (ACN; M2), 500 μL 80% ACN (M3), 500 μL 50% MeOH (M4) and 500 μL 80% MeOH (M5).…”

“…Five extraction solvents were chosen, designed and compared for liver metabolite extraction based on previous studies(Huang et al, 2011;Liu et al, 2015;Masson, Alves, Ebbels, Nicholson, & Want, 2010). Five extraction solvents were chosen, designed and compared for liver metabolite extraction based on previous studies(Huang et al, 2011;Liu et al, 2015;Masson, Alves, Ebbels, Nicholson, & Want, 2010).…”

“…. Supernatant, 5 μL, was injected for analysis after centrifugation.To examine the influence of extraction fold (tissue/mg: solution/ μL) on extracted metabolites, four folds were compared(Cai et al, 2016;Liu et al, 2015;Lu et al, 2013). Liver samples (50 mg) were first homogenized in 200 μL ice-cold water followed by addition of 300 μL methanol (MeOH; M1), 500 μL 50% acetonitrile (ACN; M2), 500 μL 80% ACN (M3), 500 μL 50% MeOH (M4) and 500 μL 80% MeOH (M5).After the supernatant (150 μL) had been diluted with 150 μL chilled 100% ACN and vortexed for 20 min, the mixture was centrifuged.Owing to supernatant frequently being treated by drying and redissolution, eight resuspension solutions were designed for comparison (Garcia-Canaveras, Donato, Castell, & Lahoz, 2011; Huang et al, 2011; Jeong et al, 2015; Masson et al, 2010).…”