title: Diclofenac in fish: blood plasma levels similar to human therapeutic levels affect global hepatic gene expression.
abstract: Diclofenac is a nonsteroidal anti-inflammatory drug frequently found in the aquatic environment. Previous studies have reported histological changes in the liver, kidney, and gills of fish at concentrations similar to those measured in treated sewage effluents (approximately 1 µg/L). Analyses or predictions of blood plasma levels in fish allow a direct comparison with human therapeutic plasma levels and may therefore be used to indicate a risk for pharmacological effects in fish. To relate internal exposure to a pharmacological interaction, we investigated global hepatic gene expression together with bioconcentration in blood plasma and liver of rainbow trout (Oncorhynchus mykiss) exposed to waterborne diclofenac. At the highest exposure concentration (81.5 µg/L), the fish plasma concentration reached approximately 88% of the human therapeutic levels (C(max) ) after two weeks. Using an oligonucleotide microarray followed by quantitative PCR, we found extensive effects on hepatic gene expression at this concentration, and some genes were found to be regulated down to the lowest exposure concentration tested (1.6 µg/L), corresponding to a plasma concentration approximately 1.5% of the human C(max) . Thus, at concentrations detected in European surface waters, diclofenac can affect the expression of multiple genes in exposed fish. Functional analysis of differentially expressed genes revealed effects on biological processes such as inflammation and the immune response, in agreement with the mode of action of diclofenac in mammals. In contrast to some previously reported results, the bioconcentration factor was found to be stable (4.02 ± 0.75 for blood plasma and 2.54 ± 0.36 for liver) regardless of the water concentration.
authors.affiliation: Sahlgrenska Academy at the University of Gothenburg, Göteborg, Sweden.
authors.forename: Filip
authors.initials: F
authors.lastname: Cuklev
authors.forename: Erik
authors.initials: E
authors.lastname: Kristiansson
authors.forename: Jerker
authors.initials: J
authors.lastname: Fick
authors.forename: Noomi
authors.initials: N
authors.lastname: Asker
authors.forename: Lars
authors.initials: L
authors.lastname: Förlin
authors.forename: D G Joakim
authors.initials: DG
authors.lastname: Larsson
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000633:toxicity*; D004008:Diclofenac / Q000097:blood / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005618:Fresh Water / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects*; D005880:Gills / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D017686:Oncorhynchus mykiss / Q000097:blood* / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018570:Risk Assessment; D012722:Sewage; D014874:Water Pollutants, Chemical / Q000097:blood / Q000633:toxicity*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D012333:RNA, Messenger; D012722:Sewage; D014874:Water Pollutants, Chemical; D004008:Diclofenac