title: Inhibition of cytochrome P450s enhances (+)-usnic acid cytotoxicity in primary cultured rat hepatocytes.
abstract: (+)-Usnic acid (UA) is consumed as a dietary supplement to promote weight loss; however, dietary supplements containing UA have been associated with clinical cases of severe liver injury. UA has been shown to be hepatotoxic in rats and is extensively metabolized by hepatic cytochrome P450s (CYPs); therefore, we examined if UA metabolism results in the formation of cytotoxic metabolites or if metabolism is a detoxification process in primary rat hepatocytes. When CYP activity was suppressed by the non-isoenzyme-selective inhibitor SKF-525A (20 μM), or the CYP1A inhibitor alpha-naphthoflavone (10 μM), or the CYP3A inhibitor ketoconazole (25 μM), the cytotoxicity of UA at 3~6 μM after 3~20 h of exposure was significantly increased as measured by lactate dehydrogenase (LDH) leakage. At 2 h after UA exposure, an earlier time point prior to LDH release, these CYP inhibitors potentiated UA-induced inhibition of cellular respiration as determined by the Clark type oxygen electrode. Cellular adenosine triphosphate (ATP) depletion by UA was also exacerbated by these CYP inhibitors. The CYP2B/2C inhibitor, ticlopidine at 20 μM, showed no effects in parallel experiments. These data demonstrate that UA is bio-transformed to less toxic metabolites in rat primary hepatocytes, probably mainly by CYP1A and 3A, but not 2B/2C. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.
authors.affiliation: Division of Systems Biology, National Center for Toxicological Research, U.S. FDA, 3900 NCTR Road, Jefferson, AR, 72079, USA.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Shi
authors.forename: James
authors.initials: J
authors.lastname: Greenhaw
authors.forename: William F
authors.initials: WF
authors.lastname: Salminen
mesh_terms: D000818:Animals; D001571:Benzoflavones / Q000737:chemistry; D001572:Benzofurans / Q000009:adverse effects*; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000473:pathology; D065607:Cytochrome P-450 Enzyme Inhibitors / Q000737:chemistry*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D019587:Dietary Supplements; D004791:Enzyme Inhibitors / Q000737:chemistry; D022781:Hepatocytes / Q000187:drug effects*; D007654:Ketoconazole / Q000737:chemistry; D008099:Liver / Q000187:drug effects / Q000473:pathology; D008297:Male; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D011335:Proadifen / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D013988:Ticlopidine / Q000737:chemistry
chemical_list: D001571:Benzoflavones; D001572:Benzofurans; D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; C073339:usnic acid; C011512:alpha-naphthoflavone; D003577:Cytochrome P-450 Enzyme System; D011335:Proadifen; D013988:Ticlopidine; D007654:Ketoconazole
keywords: cytochrome P450; hepatocytes; hepatotoxicity; rat; usnic acid