Analysis of the Effects Diclofenac has on Japanese Medaka (Oryzias latipes) Using Real-time PCR

National Research Laboratory on Environmental Biotechnology, Gwangju Institute of Science and Technology (GIST), Gwangju, South Korea.
Chemosphere (Impact Factor: 3.34). 06/2007; 67(11):2115-21. DOI: 10.1016/j.chemosphere.2006.12.090
Source: PubMed


The expression levels of cytochrome P450 1A, p53 and vitellogenin were investigated in three different tissues of male medaka fish after exposure to diclofenac that is one of the main concerns among pharmaceuticals frequently found in sewage treatment plant (STP) effluents. The results showed that cytochrome P450 1A, p53 and vitellogenin were highly expressed in tissue-specific gene expression patterns after exposure to 8 mg/l and 1 microg/l of diclofenac. These elevated expression levels of three biomarkers suggested that diclofenac has potential to cause cellular toxicity, p53-related genotoxicity and estrogenic effects. It is also noteworthy that diclofenac has the potential to cause these effects even at an environmentally relevant concentration of diclofenac, 1 microg/l.

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    • "Using data presented in multiple studies we reviewed, we analyzed the relationship between intersex prevalence from multiple fish families and contaminant data for a number chemicals for which these data were available using weighted regressions (weighted by sample size). We observed one positive relationship between the concentration of a xenobiotic, diclofenac; a non-steroidal anti-inflammatory drug with endocrine-disrupting potential (Hong et al., 2007), and the prevalence of intersex (measured as the percentage of fish sampled with intersex lesions; R 2 = 39, P = 0.05, n = 10 studies), and one marginally significant negative relationship between dichlorodiphenyldichloroethylene (DDE), a metabolite of dichlorodiphenyltrichloroethane (DDT) concentration and the prevalence of intersex (R 2 = 60, P = 0.07, n = 6 studies). We did not, however, observe a significant relationship between the prevalence of intersex and concentration of atrazine (R 2 = 20, P = 0.38, n = 6 studies), an estrogenic contaminant . "
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    ABSTRACT: Intersex is defined as the abnormal presence of both testicular and ovarian cells in gonads of gonochoristic animals. Its occurrence is widespread and reports on its presence in the gonads of vertebrates continues to increase. In this review, we use standardized terminology to summarize the current knowledge of intersex in gonochoristic fishes and amphibians. We describe the different indices that have been used to assess the severity of intersex and synthesize reports discussing the prevalence of intersex in relation to different types of pollutants. In addition, we evaluate the geographic distribution and chronology of the reported cases of intersex in fishes and amphibians, their pathological descriptions and severity and discuss species sensitivities. We also summarize molecular biomarkers that have been tested for early detection of intersex in wild populations and highlight additional biomarkers that target molecular pathways involved in gonadal development that require further investigation for use in the diagnosis of intersex. Finally, we discuss the needs for future research in this field. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.
    Journal of Applied Toxicology 07/2015; DOI:10.1002/jat.3204 · 2.98 Impact Factor
    • "The liver is the primary organ of detoxification of xenobiotics. Although, the intestine is the major organ of digestion and absorption, several biotransformation enzymes involved in detoxification and elimination are located in the fish intestine (Hong et al., 2007). 4.1. "
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    ABSTRACT: The aim of this study was to investigate the effects of naproxen on the gene expression of antioxidant enzymes in adult zebrafish. Surprisingly, after 2 weeks exposure no significant effect on the mRNA expression of the target genes was found in the liver. However, mRNA levels of three genes were altered significantly in the intestine. The expression of Ucp-2 decreased at the environmental concentration of 1μg/L while mRNA expression of GST p2 increased at the concentration of 100μg/L. The mRNA level for the antioxidant enzyme CAT was up-regulated significantly at both the concentrations used. Exposure to naproxen caused only moderate effects on the expression of antioxidant genes in the intestine rather than in the liver, which demonstrates that the intestine is more sensitive to waterborne naproxen exposure than the liver. Interestingly, the adverse side effects of NSAIDs occur in the gastrointestinal tract of humans. To our knowledge, this is the first study that has focused on transcriptional effects of naproxen on zebrafish. Copyright © 2015 Elsevier B.V. All rights reserved.
    07/2015; 40(2):343-348. DOI:10.1016/j.etap.2015.07.009
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    • "CYP1A is a mixed function oxidase that plays an important role in the phase I biotransformation of xenobiotics (Andersson and Forlin, 1992). In fish, transcript levels of cyp1a have previously been shown to increase during exposure to diclofenac (Hong et al., 2007; Mehinto et al., 2010). In addition, several other pharmaceuticals have been shown to induce or inhibit CYP1A enzyme activity, measured as either ethoxyresorufin-O-deethylase (EROD) activity or gene transcription, suggesting CYP1A may have a significant role in their metabolism (Bartram et al., 2012; Beijer et al., 2013; Fernandez et al., 2013). "
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    ABSTRACT: Pollution with low concentrations of pharmaceuticals, especially when combined with low-oxygen conditions (hypoxia), is a threat to aquatic ecosystems worldwide. The non-steroidal anti-inflammatory drug diclofenac is commonly detected in wastewater effluents, and has potential to accumulate in the bile of fish. Diclofenac has been shown to activate aryl hydrocarbon receptor (AHR), which induces transcription in the metabolic enzyme cytochrome P450 1a (cyp1a). Previously, crosstalk has been shown to occur between AHR and hypoxia inducible factor 1 (HIF-1). In addition, both of these transcription factors interact with the proteins regulating circadian (24-h) rhythms in vertebrates. Yet little is known about the significance of these interactions during simultaneous exposure to chemicals and hypoxia in fish in vivo. We exposed wild-caught three-spined sticklebacks (Gasterosteus aculeatus) to diclofenac (1 μg/L, 14 days), hypoxia (2.0 mg/L, up to 24 h) and the combination of both. We then analyzed markers of chemical biotransformation (EROD activity, cyp1a and ahr mRNA levels), glycolysis (lactate dehydrogenase (LDH) enzyme activity, ldh and enolase 1a mRNA levels), and the transcription of core circadian clock genes clock and period 1 in liver tissue. Samples were taken at three time points during the light period in order to address disturbances in the circadian variation of metabolic processes. The results show that mRNA levels and LDH activity tended to be lowest before the dark period, but this pattern was disturbed by hypoxia and diclofenac. Diclofenac and hypoxia co-exposure induced EROD activity more strongly than diclofenac exposure alone, while cyp1a mRNA level was increased also by hypoxia and diclofenac alone. LDH activity and mRNA expression showed a clear time-dependent response during hypoxia, which is consistent with the previously suggested decreased accumulation of HIF-1 during the dark period. Furthermore, LDH activity and transcription was disturbed by diclofenac, indicating important effects of environmental pollutants in disturbing natural acclimation. This study demonstrates the need for more studies to understand the potential disturbances in endogenous rhythms caused by environmental pollution in natural populations. Full text available from,OE92DAF
    Aquatic Toxicology 01/2015; 158(116):124. DOI:10.1016/j.aquatox.2014.11.006 · 3.45 Impact Factor
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