title: Effects of cytochrome P450 1A substrate (difloxacin) on enzyme gene expression and pharmacokinetics in crucian carp (hybridized Prussian carp).
abstract: Cytochrome P450s (CYPs) play a prominent role in drug metabolism and biotransformation which are distributed in liver of aquatic animals. However, limited information is available about CYP genes involved in drug metabolism in fish. In the present study, we explore CYP1A characterization for DIF metabolism. Firstly, we cloned and characterized the full-length cDNA sequence of a CYP1A gene from crucian carp (hybridized Prussian carp), the predicted protein sequence for CYP1A comprise 496 amino acids. The heme-binding region of the CYP1A, encompassing the amino acid sequence GLGKRRCIG, which is identical to the same region of other homologues. Secondly, we studied the difloxacin (DIF) kinetics and the effects of DIF on their corresponding CYP1A mRNA levels in liver of crucian carp. CYP1A1 mRNA expression was analyzed by real-time PCR, and DIF concentration was determined by reversed-phase high-performance liquid chromatography (RP-HPLC). Results showed that the concentration of DIF in liver reached its peak (67.70 mg kg(-1)) at 0.5h, while the CYP1A1 gene expression was at the lowest point. CYP1A mRNA was down-regulated by 6.5 mg ml(-1) DIF in the liver of crucian carp. Thus, our work confirmed that DIF is both the substrate and inhibitor of CYP1A. The information provided a model for the potential utility of gene expression analysis and drug metabolization in fish.
authors.affiliation: National Center for Aquatic Pathogen Collection, College of Fisheries and Life Science, Shanghai Ocean University, 999 Hucheng Huan Road, Shanghai 201306, China.
authors.forename: Gui Hong
authors.initials: GH
authors.lastname: Fu
authors.forename: Xian Le
authors.initials: XL
authors.lastname: Yang
authors.forename: Hai Xin
authors.initials: HX
authors.lastname: Zhang
authors.forename: Wen Juan
authors.initials: WJ
authors.lastname: Yu
authors.forename: Kun
authors.initials: K
authors.lastname: Hu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000097:blood / Q000493:pharmacokinetics / Q000494:pharmacology*; D001483:Base Sequence; D002347:Carps / Q000235:genetics* / Q000378:metabolism; D002939:Ciprofloxacin / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics / Q000494:pharmacology; D003001:Cloning, Molecular; D019363:Cytochrome P-450 CYP1A1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D004791:Enzyme Inhibitors / Q000097:blood / Q000493:pharmacokinetics / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006207:Half-Life; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D014018:Tissue Distribution
chemical_list: D000900:Anti-Bacterial Agents; D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D002939:Ciprofloxacin; C047225:difloxacin; D019363:Cytochrome P-450 CYP1A1