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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    • "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.
    No preview · Article · Jul 2015
    • "NSAIDs induce both geno-and cytotoxicity on aquatic organisms such as Oryzias latipes, Dreissena polymorpha, Ruditapes philippinarum and Daphnia magna (Gómez-Oliván et al., 2013; Hong et al., 2007; Matozzo et al., 2012; Parolini et al., 2010). In addition, pharmaceutical effluents that contain analgesics have been shown to induce genotoxicity in Allium cepa and Swiss mice (Bakare et al., 2009; Oliveira-Júnior et al., 2013). "
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    ABSTRACT: The pharmaceutical industry generates wastewater discharges of varying characteristics and contaminant concentrations depending on the nature of the production process. The main chemicals present in these effluents are solvents, detergents, disinfectants - such as sodium hypochlorite (NaClO) - and pharmaceutical products, all of which are potentially ecotoxic. Therefore, this study aimed to evaluate the geno- and cytotoxicity induced in the common carp Cyprinus carpio by the effluent emanating from a nonsteroidal anti-inflammatory drug (NSAID)-manufacturing plant. Carp were exposed to the lowest observed adverse effect level (LOAEL, 0.1173%) for 12, 24, 48, 72 and 96h, and biomarkers of genotoxicity (comet assay and micronucleus test) and cytotoxicity (caspase-3 activity and TUNEL assay) were evaluated. A significant increase with respect to the control group (p<0.05) occurred with all biomarkers from 24h on. Significant positive correlations were found between NSAID concentrations and biomarkers of geno- and cytotoxicity, as well as among geno- and cytotoxicity biomarkers. In conclusion, exposure to this industrial effluent induces geno- and cytotoxicity in blood of C. carpio. Copyright © 2015 Elsevier B.V. All rights reserved.
    No preview · Article · May 2015 · Science of The Total Environment
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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
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