title: Evidence that unsaturated fatty acids are potent inhibitors of renal UDP-glucuronosyltransferases (UGT): kinetic studies using human kidney cortical microsomes and recombinant UGT1A9 and UGT2B7.
abstract: Renal ischaemia is associated with accumulation of fatty acids (FA) and mobilisation of arachidonic acid (AA). Given the capacity of UDP-glucuronosyltransferase (UGT) isoforms to metabolise both drugs and FA, we hypothesised that FA would inhibit renal drug glucuronidation. The effect of FA (C2:0-C20:5) on 4-methylumbelliferone (4-MU) glucuronidation was investigated using human kidney cortical microsomes (HKCM) and recombinant UGT1A9 and UGT2B7 as the enzyme sources. 4-MU glucuronidation exhibited Michaelis-Menten kinetics with HKCM (apparent K(m) (K(m)(app)) 20.3 microM), weak substrate inhibition with UGT1A9 (K(m)(app) 10.2 microM, K(si) 289.6 microM), and sigmoid kinetics with UGT2B7 (S(50)(app)440.6 microM) Similarly, biphasic UDP-glucuronic acid (UDPGA) kinetics were observed with HKCM (S(50) 354.3 microM) and UGT1A9 (S(50) 88.2 microM). In contrast, the Michaelis-Menten kinetics for UDPGA observed with UGT2B7 (K(m)(app) 493.2 microM) suggested that kinetic interactions with UGTs were specific to the xenobiotic substrate and the co-substrate (UDPGA). FA (C16:1-C20:5) significantly inhibited (25-93%) HKCM, UGT1A9 or UGT2B7 catalysed 4-MU glucuronidation. Although linoleic acid (LA) and AA were both competitive inhibitors of 4-MU glucuronidation by HKCM (K(i)(app) 6.34 and 0.15 microM, respectively), only LA was a competitive inhibitor of UGT1A9 (K(i)(app) 4.06 microM). In contrast, inhibition of UGT1A9 by AA exhibited atypical kinetics. These data indicate that LA and AA are potent inhibitors of 4-MU glucuronidation catalysed by human kidney UGTs and recombinant UGT1A9 and UGT2B7. It is conceivable therefore that during periods of renal ischaemia FA may impair renal drug glucuronidation thus compromising the protective capacity of the kidney against drug-induced nephrotoxicity.
authors.affiliation: Department of Clinical Pharmacology, Flinders University and Flinders Medical Centre, Bedford Park, 5042, Adelaide, Australia.
authors.forename: Paraskevi
authors.initials: P
authors.lastname: Tsoutsikos
authors.forename: John O
authors.initials: JO
authors.lastname: Miners
authors.forename: Alan
authors.initials: A
authors.lastname: Stapleton
authors.forename: Anthony
authors.initials: A
authors.lastname: Thomas
authors.forename: Benedetta C
authors.initials: BC
authors.lastname: Sallustio
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Knights
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D014453:Glucuronosyltransferase / Q000037:antagonists & inhibitors* / Q000235:genetics; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000378:metabolism; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008297:Male; D008861:Microsomes / Q000187:drug effects* / Q000201:enzymology; D008875:Middle Aged; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D000090264:UDP-Glucuronosyltransferase 1A9; D014535:Uridine Diphosphate Glucuronic Acid / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D005231:Fatty Acids, Unsaturated; D011994:Recombinant Proteins; C000715591:UGT1A9 protein, human; C019492:4-methylumbelliferyl sulfate; D014535:Uridine Diphosphate Glucuronic Acid; D006923:Hymecromone; C491474:UGT2B7 protein, human; D014453:Glucuronosyltransferase; D000090264:UDP-Glucuronosyltransferase 1A9