The effects of the painkiller diclofenac and hypoxia on gene transcription and antioxidant system in the gills of three-spined stickleback.
Aquatic organisms face multiple stressors in natural ecosystems. More and more often painkillers are detected in surface waters since their prescription has increased worldwide within the last years. Here we examined the effects of the non-steroidal anti-inflammatory drug (NSAID) diclofenac and hypoxia on three-spined sticklebacks (Gasterosteus aculeatus). We exposed sticklebacks to an environmentally relevant concentration of diclofenac (1μg/L) for 14days, to 24h of hypoxia (2.0mg O2/L), and a combination of both. Hypoxia and diclofenac both can be associated with oxidative stress in fish, but it is unclear whether they would act synergistically. Expression analysis of genes related to antioxidant response, hypoxia response, and chemical metabolism in gills showed that diclofenac alone had little effect, while the combination of hypoxia and diclofenac affected transcript levels most, indicating synergistic effects of these stressors. Of the antioxidant enzymes, only superoxide dismutase activity remained unchanged by treatments, while glutathione peroxidase (GPx) was the most affected antioxidant response on both the transcript and activity levels. Our results suggest that diclofenac may lead to suppressed catalase (CAT) activity but increased GPx activity, probably as compensatory mechanism to remove increasing H2O2 in the gills, and that this response is not affected by hypoxia. The activities of lactate dehydrogenase, CAT, and GPx also showed temporal variability during treatments, which can be attributable to tissue-specific circadian rhythms. Our study shows how responses to NSAIDs and hypoxia can interact in fish, suggesting that getting more insight into temporal variation and about the different levels of regulation of environmental responses is necessary in future studies.
Zoological Institute and Zoological Museum, Biocenter Grindel, University of Hamburg, D-20146 Hamburg, Germany.
Pedro
P
Lubiana
Laboratory of Animal Physiology, Department of Biology, University of Turku, Turku FIN-20014, Finland.
Jenni M
JM
Prokkola
Laboratory of Animal Physiology, Department of Biology, University of Turku, Turku FIN-20014, Finland.
Mikko
M
Nikinmaa
Zoological Institute and Zoological Museum, Biocenter Grindel, University of Hamburg, D-20146 Hamburg, Germany.
Thorsten
T
Burmester
Laboratory of Animal Physiology, Department of Biology, University of Turku, Turku FIN-20014, Finland.
Mirella
M
Kanerva
Zoological Institute and Zoological Museum, Biocenter Grindel, University of Hamburg, D-20146 Hamburg, Germany; Laboratory of Animal Physiology, Department of Biology, University of Turku, Turku FIN-20014, Finland. Electronic address: miriam.gotting@utu.fi.
Miriam
M
Götting
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000633:toxicity*; D000975:Antioxidants / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D015687:Cell Hypoxia; D004008:Diclofenac / Q000633:toxicity*; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D005880:Gills / Q000187:drug effects* / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000235:genetics / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D023701:Smegmamorpha / Q000235:genetics / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D000975:Antioxidants; D015415:Biomarkers; D029941:Fish Proteins; D004008:Diclofenac; D007770:L-Lactate Dehydrogenase; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase
Ecotoxicology; Fish; Gasterosteus aculeatus; Gills; Hypoxia; Oxidative stress; Pharmaceuticals