title: Copper metabolism and oxidative stress in chronic inflammatory and cholestatic liver diseases in dogs.
abstract: Inherited defects of copper metabolism resulting in hepatic copper accumulation and oxidative-stress might cause breed-associated forms of hepatitis. Biliary excretion is the major elimination route of copper, therefore increased hepatic copper concentrations could also be caused by cholestasis. The aim of this study was to find criteria to determine whether copper-accumulation is primary or occurs secondary to hepatitis. Liver samples of Bedlington Terriers with copper toxicosis (CT), breeds with non-copper-associated chronic extrahepatic cholestasis (EC) or chronic hepatitis (CH), and healthy dogs were used. Copper metabolism was analyzed by means of histochemical staining (copper concentration) and quantitative reverse transcriptase polymerase chain reaction (Q-PCR) on copper excretion/storage (ATOX1, COX17, ATP7A, ATP7B, CP, MT1A, MURR1, XIAP). Oxidative stress was measured by determining GSH/GSSG ratios and gene-expression (SOD1, CAT, GSHS, GPX1, CCS, p27KIP, Bcl-2). Results revealed 5+ copper in CT, but no or 1-2+ copper in EC and CH. Most gene products for copper metabolism remained at concentrations similar to healthy dogs. Three clear exceptions were observed in CT: 3-fold mRNA increase of ATP7A and XIAP and complete absence of MURRI. The only quantitative differences between the diseased and the control groups were in oxidative stress, evidenced by reductions in all GSH/GSSG ratios. We conclude that 3+ or higher histochemical detection of copper indicates a primary copper storage disease. The expression profile of copper-associated genes can be used as a reference for future studies on copper-associated diseases. All 3 diseases have reduced protection against oxidative stress, opening a rationale to use antioxidants as possible therapy.
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, The Netherlands. B.Spee@vet.uu.nl
authors.forename: Bart
authors.initials: B
authors.lastname: Spee
authors.forename: Brigitte
authors.initials: B
authors.lastname: Arends
authors.forename: Ted S G A M
authors.initials: TS
authors.lastname: van den Ingh
authors.forename: Louis C
authors.initials: LC
authors.lastname: Penning
authors.forename: Jan
authors.initials: J
authors.lastname: Rothuizen
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001706:Biopsy / Q000662:veterinary; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D001651:Cholestasis, Extrahepatic / Q000378:metabolism / Q000473:pathology / Q000662:veterinary*; D003300:Copper / Q000378:metabolism*; D004283:Dog Diseases / Q000378:metabolism* / Q000473:pathology; D004285:Dogs; D005978:Glutathione / Q000378:metabolism; D006521:Hepatitis, Chronic / Q000378:metabolism / Q000473:pathology / Q000662:veterinary*; D006651:Histocytochemistry / Q000662:veterinary; D018384:Oxidative Stress / Q000502:physiology*; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013854:Thioamides / Q000737:chemistry
chemical_list: D000975:Antioxidants; D027682:Cation Transport Proteins; D013854:Thioamides; D012313:RNA; D003300:Copper; D005978:Glutathione; C005728:rubeanic acid