title: ANIT-induced intrahepatic cholestasis alters hepatobiliary transporter expression via Nrf2-dependent and independent signaling.
abstract: Alpha-naphthylisothiocyanate (ANIT) causes intrahepatic cholestasis by injuring biliary epithelial cells. Adaptive regulation of hepatobiliary transporter expression has been proposed to reduce liver injury during cholestasis. Recently, the oxidative stress transcription factor Nrf2 (nf-e2-related factor 2) was shown to regulate expression of hepatobiliary transporters. The purpose of this study was to determine whether ANIT-induced hepatotoxicity and regulation of hepatobiliary transporters are altered in the absence of Nrf2. For this purpose, wild-type and Nrf2-null mice were administered ANIT (75 mg/kg po). Surprisingly, ANIT-induced hepatotoxicity was similar in both genotypes at 48 h. Accumulation of bile acids in serum and liver was lower in Nrf2-null mice compared with wild-types treated with ANIT. Transporter mRNA profiles differed between wild-type and Nrf2-null mice after ANIT. Bsep (bile salt export pump), Mdr2 (multidrug resistance gene), and Mrp3 (multidrug resistance-associated protein) efflux transporters were increased by ANIT in wild-type, but not in Nrf2-null mice. In contrast, mRNA expression of two hepatic uptake transporters, Ntcp (sodium-taurocholate cotransporting polypeptide) and Oatp1b2 (organic anion transporting peptide), were decreased in both genotypes after ANIT, with larger declines in Nrf2-null mice. mRNA expression of the transcriptional repressor of Ntcp, small heterodimeric partner (SHP), was increased in Nrf2-null mice after ANIT. Furthermore, hepatocyte nuclear factor 1alpha (HNF1alpha), which regulates Oatp1b2, was downregulated in ANIT-treated Nrf2-null mice. Preferential upregulation of SHP and downregulation of HNF1alpha and uptake transporters likely explains why Nrf2-null mice exhibited similar injury to wild-types after ANIT. A subsequent study revealed that treatment of mice with the Nrf2 activator oltipraz protects against ANIT-induced histological injury. Despite compensatory changes in Nrf2-null mice to limit ANIT toxicity, pharmacological activation of Nrf2 may represent a therapeutic option for intrahepatic cholestasis.
authors.affiliation: Department of Pharmacology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Lauren M
authors.initials: LM
authors.lastname: Aleksunes
authors.forename: Yue Julia
authors.initials: YJ
authors.lastname: Cui
authors.forename: Curtis D
authors.initials: CD
authors.lastname: Klaassen
mesh_terms: D015058:1-Naphthylisothiocyanate / Q000633:toxicity*; D000818:Animals; D001646:Bile / Q000378:metabolism*; D001647:Bile Acids and Salts / Q000378:metabolism; D001663:Bilirubin / Q000378:metabolism; D015153:Blotting, Western; D002352:Carrier Proteins / Q000096:biosynthesis*; D002780:Cholestasis, Intrahepatic / Q000139:chemically induced* / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D007202:Indicators and Reagents; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008111:Liver Function Tests; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D051267:NF-E2-Related Factor 2 / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D001647:Bile Acids and Salts; D002352:Carrier Proteins; D007202:Indicators and Reagents; D051267:NF-E2-Related Factor 2; D015058:1-Naphthylisothiocyanate; D012313:RNA; D001663:Bilirubin