Skip to main content
SpringerLink
Log in

Differential expression of HO-1 and CYP1A2 during up-regulation of ERK in stressed fish hepatocytes

  • Published:
Environmental Monitoring and Assessment Aims and scope Submit manuscript

Abstract

Induction of heme oxygenase-1 (HO-1) and extracellular signal-regulated kinase (ERK) is a crucial step in the cellular response to oxidative stress. Altered expression of such proteins in response to stress conditions is a key factor for the maintenance of cellular integrity and survival. Hypoxia-inducible factor-1 (HIF-1) and cytochrome P450 1A (CYP1A) are critical indicators of environmental exposure to hypoxia. The current study was aimed to analyse the expression pattern of HO-1, p-ERK, HIF-1α and CYP1A2 in hepatocytes of Mugil cephalus inhabiting Kovalam (unpolluted site) and Ennore (polluted site) estuaries. The impact of stress on hepatocytes was assessed by measuring the level of lipid hydroperoxides (LHP) and reduced glutathione (GSH) in hepatocytes of M. cephalus inhabiting these estuaries. The expression of HO-1, p-ERK, HIF-1α and CYP1A2 was analysed by ELISA and immunoblot analysis, respectively. There was significant decrease of GSH (19 %) and CYP1A2 (34 %) along with significant increase of LHP (31 %), HO-1 (96 %), p-ERK (p < 0.01) and HIF-1α (41 %) in hepatocytes of M. cephalus inhabiting Ennore estuary than Kovalam estuary. The present study shows that increased synthesis of HO-1 mainly regulated by p-ERK and HIF-1α may significantly contribute to the environmental adaptation processes of grey mullet, by protecting the cells from oxidative stress and stress-induced degenerative changes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Abraham, N. G., & Kappas, A. (2005). Heme oxygenase and the cardiovascular-renal system. Free Radical Biology and Medicine, 39, 1–25.

    Article  CAS  Google Scholar 

  • Adams, S. M., Giesy, J. P., Tremblay, L. A., & Eason, C. T. (2001). The use of biomarkers in ecological risk assessment: recommendations from the Christchurch conference on Biomarkers in Ecotoxicology. Biomarkers, 6, 1–6.

    Article  CAS  Google Scholar 

  • Akagi, R., Takahashi, T., & Sassa, S. (2002). Fundamental role of heme oxygenase ischemic acute renal failure. Japanese Journal of Pharmacology, 88, 127–132.

    Article  CAS  Google Scholar 

  • Aragones, J., Jones, D. R., Martin, S., Juan, M. A. S., Alfranca, A., Vidal, F., Vara, A., Merida, I., & Landazuri, M. O. (2001). Evidence for the involvement of diacylglycerol kinase in the activation of hypoxia-inducible transcription factor 1 by low oxygen tension. Journal of Biological Chemistry, 276, 10548–10555.

    Article  CAS  Google Scholar 

  • Barker, C. W., Fagan, J. B., & Pasco, D. S. (1994). Down-regulation of P4501A1 and P4501A2 mRNA expression in isolated hepatocytes by oxidative stress. Journal of Biological Chemistry, 269, 3985–3990.

    CAS  Google Scholar 

  • Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Analytical Biochemistry, 72, 248–254.

    Article  CAS  Google Scholar 

  • Buckley, B. A., Place, S. P., & Hofmann, G. E. (2004). Regulation of heat shock genes in isolated hepatocytes from an Antarctic fish, Trematomus bernacchii. Journal of Experimental Biology, 207, 3649–3656.

    Article  CAS  Google Scholar 

  • Buet, A., Barilloet, S., & Camilleri, V. (2002). Changes in oxidative stress parameters in fish as response to direct uranium exposure. Radio Protect Supply, 1(40), S151–S155.

    Google Scholar 

  • Chan, W. K., Yao, G., Gu, Y. Z., & Bradfield, C. A. (1999). Cross-talk between the aryl hydrocarbon receptor and hypoxia inducible factor signaling pathways. Demonstration of competition and compensation. Journal of Biological Chemistry, 274, 12115–12123.

    Article  CAS  Google Scholar 

  • Chen, K., & Maines, M. D. (2000). Nitric oxide induces heme oxygenase-1 via mitogen-activated protein kinases ERK and p38. Cellular and Molecular Biology, 46(3), 609–617.

    CAS  Google Scholar 

  • Chen, J. C., Huang, K. C., & Lin, W. W. (2006). HMG-CoA reductase inhibitors upregulate heme oxygenase-1 expression in murine RAW264.7 macrophages via ERK, p38 MAPK and protein kinase G pathways. Cellular Signalling, 18, 32–39.

    Article  CAS  Google Scholar 

  • Choi, A., & Alam, J. (1996). Heme oxygenase-1: function, regulation, and implication of a novel stress-inducible protein in oxidant-induced lung injury. American Journal of Respiratory Cell and Molecular Biology, 15, 9–19.

    Article  CAS  Google Scholar 

  • Dennery, P. A. (2000). Regulation and role of heme oxygenase in oxidative injury. Current Topics in Cellular Regulation, 36, 181–199.

    Article  CAS  Google Scholar 

  • Di-Giulio, R. T., Benson, W. H., Sanders, B. M., & van Veld, P. A. (1995). Biochemical mechanisms: metabolism, adaptation, and toxicity. In G. M. Rand (Ed.), Fundamentals of aquatic toxicology: effects, environmental fate, and risk assessment (2nd ed., pp. 523–562). London: Taylor and Francis.

    Google Scholar 

  • Dimova, E. Y., Michiels, C., & Kietzmann, T. (2009). Kinases as upstream regulators of the HIF system: their emerging potential as anti-cancer drug targets. Current Pharmaceutical Design, 15(33), 3867–3877.

    Article  CAS  Google Scholar 

  • El-Kadi, A. O. S., Bleau, A. M., Dumont, I., Maurice, H., & du Souich, P. (2000). Role of reactive oxygen intermediates in the decrease of hepatic cytochrome P450 activity by serum of human and rabbits with an acute inflammatory reaction. Drug Metabolism and Disposition, 28, 1112–1120.

    CAS  Google Scholar 

  • Farombi, E. O., & Surh, Y. J. (2006). Heme oxygenase-1 as a potential therapeutic target for hepatoprotection. Journal of Biochemistry and Molecular Biology, 39(5), 479–491.

    Article  CAS  Google Scholar 

  • Fischer, W., Bianchi, G. (1984). FAO species identification sheets for fishery purposes, Western Indian Ocean (fishing area 51), FAO, Rome, Vol. I-6.

  • Fleming, C. R., & Di-Giulio, R. T. (2011). The role of CYP1A inhibition in the embryotoxic interactions between hypoxia and polycyclic aromatic hydrocarbons (PAHs) and PAH mixtures in zebrafish (Danio rerio). Ecotoxicology, 20, 1300–1314.

    Article  CAS  Google Scholar 

  • Fogg, S., Agarwal, A., Nick, H. S., & Visner, G. A. (1999). Iron regulates hyperoxia-dependent human heme oxygenase 1 gene expression in pulmonary endothelial cells. American Journal of Respiratory Cell and Molecular Biology, 20, 797–804.

    Article  CAS  Google Scholar 

  • Fradette, C., & du Souich, P. (2004). Effect of hypoxia on cytochrome P450 activity and expression. Current Drug Metabolism, 5, 257–271.

    Article  CAS  Google Scholar 

  • Fradette, C., Batonga, J., Teng, S., Piquette-Miller, M., & du Souich, P. (2007). Animal models of acute moderate hypoxia are associated with a down-regulation of CYP1A1, 1A2, 2B4, 2C5, and 2C16 and up-regulation of CYP3A6 and P-glycoprotein in liver. Drug Metabolism and Disposition, 35, 765–771.

    Article  CAS  Google Scholar 

  • Gernhofer, M., Pawet, M., Schramm, M., Muller, E., & Triebskorn, R. (2001). Ultrastructural biomarkers as tools to characterize the health status of fish in contaminated streams. Journal of Aquatic Ecosystem Stress and Recovery, 8, 241–260.

    Article  CAS  Google Scholar 

  • Goksoyr, A., Husoy, A., Larsen, H., Klungsoyr, J., Wilhelmsen, S., Maage, A., Brevik, E., Andersson, T., Celander, M., Pesonen, M., & Forlin, L. (1991). Environmental contaminants and biochemical responses in flatfish from the Hvaler Archipelago in Norway. Archives of Environmental Contamination and Toxicology, 21, 486–496.

    Article  CAS  Google Scholar 

  • Gong, P., Cederbaum, A. I., & Nieto, N. (2004). Heme oxygenase-1 protects HepG2 cells against cytochrome P450 2E1-dependent toxicity. Free Radical Biology and Medicine, 36(3), 307–318.

    Article  CAS  Google Scholar 

  • Ikeyama, S., Kokkonen, G., Shack, S., Wang, X. T., & Holbrook, N. J. (2002). Loss in oxidative stress tolerance with aging linked to reduced extracellular signal-regulated kinase and Akt kinase activities. FASEB Journal, 16, 114–116.

    CAS  Google Scholar 

  • Jiang, Z. Y., Hunt, J. V., & Wolff, S. P. (1992). Ferrous ion oxidation in the presence of xylenol orange for detection of lipid hydroperoxide in low density lipoprotein. Analytical Biochemistry, 202, 384–389.

    Article  CAS  Google Scholar 

  • Kaluz, S., Kaluzova, M., & Stanbridge, E. J. (2006). The role of extracellular signal-regulated protein kinase in transcriptional regulation of the hypoxia marker carbonic anhydrase IX. Journal of Cellular Biochemistry, 97, 207–216.

    Article  CAS  Google Scholar 

  • Kaplowitz, N., Aw, T. Y., & Ookhtens, M. (1985). The regulation of hepatic glutathione. Annual Review of Pharmacology and Toxicology, 25, 715–744.

    Article  CAS  Google Scholar 

  • Krumschnabel, G., Schwarzbaum, P. J., & Wieser, W. (1994). Coupling of energy supply and energy demand in isolated goldfish hepatocytes. Physiological Zoology, 67, 438–448.

    Google Scholar 

  • Laville, N., Ait-Aissa, S., Gomez, E., Casellas, C., & Porcher, J. M. (2004). Effects of human pharmaceuticals on cytotoxicity, EROD activity and ROS production in fish hepatocytes. Toxicology, 196(1–2), 41–55.

    Article  CAS  Google Scholar 

  • Lee, P. J., Jiang, B. H., Chin, B. Y., Iyer, N. V., Alam, J., Semenza, G. L., & Choi, A. M. K. (1997). Hypoxia-inducible factor-1 mediates transcriptional activation of the heme oxygenase-1 gene in response to hypoxia. Journal of Biological Chemistry, 272, 5375–5381.

    Article  CAS  Google Scholar 

  • Lee, S. M., Lee, C. T., Kim, Y. W., Han, S. K., Shim, Y. S., & Yoo, C. G. (2006). Hypoxia confers protection against apoptosis via PI3K/Akt and ERK pathways in lung cancer cells. Cancer Letters, 242, 231–238.

    Article  CAS  Google Scholar 

  • Lewis, D. F. V., Watson, E., & Lake, B. G. (1998). Evolution of the cytochrome P450 superfamily: sequence alignments and pharmacogenetics. Mutation Research, 410, 245–270.

    Article  CAS  Google Scholar 

  • Li, B., Takeda, K., Yokoyama, S., & Shibahara, S. (2008). A prolyl-hydroxylase inhibitor, ethyl-3,4-dihydroxybenzoate, induces haem oxygenase-1 expression in human cells through a mechanism independent of hypoxia-inducible factor-1alpha. Journal of Biochemistry, 144, 643–654.

    Article  CAS  Google Scholar 

  • Livingstone, D. R. (2001). Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Marine Pollution Bulletin, 42, 656–666.

    Article  CAS  Google Scholar 

  • McGlashan, D. J., & Hughies, J. M. (2001). Genetic evidence for historical continuity between populations of the Australian freshwater fish Craterocephalus stercusmuscarum (Atherinidae) east and west of the Great Diving Range. Journal of Fish Biology, 59, 55–67.

    Article  CAS  Google Scholar 

  • Moron, M. S., Beperre, J. W., & Wick, B. (1979). Levels of glutathione, glutathione reductase, glutathione-stransferase in rat lung and liver. Biochimica et Biophysica Acta, 582, 67–78.

    Article  CAS  Google Scholar 

  • Mylonis, I., Chachami, G., Samiotaki, M., Panayotou, G., Paraskeva, E., Kalousi, A., Georgatsou, E., Bonanou, S., & Simos, G. (2006). Identification of MAPK phosphorylation sites and their role in the localization and activity of hypoxiainducible factor-1alpha. Journal of Biological Chemistry, 281(44), 33095–33106.

    Article  CAS  Google Scholar 

  • Ogborne, R. M., Rushworth, S. A., & O’Connell, M. A. (2005). α-Lipoic acid-induced heme oxygenase-1 expression is mediated by nuclear factor erythroid 2-related factor 2 and p38 mitogen-activated protein kinase in human monocytic cells. Arteriosclerosis, Thrombosis, and Vascular Biology, 25, 2100–2105.

    Article  CAS  Google Scholar 

  • Olsvik, P. A., Kristensen, T., Waagbo, R., Tollefsen, K. E., Rosseland, B. O., & Toften, H. (2006). Effects of hypo- and hyperoxia on transcription levels of five stress genes and the glutathione system in liver of Atlantic cod Gadus morhua. Journal of Experimental Biology, 209, 2893–2901.

    Article  CAS  Google Scholar 

  • Padmini, E., & Usha Rani, M. (2008). Impact of seasonal variation on HSP70 expression quantitated in stressed fish hepatocytes. Comparative Biochemistry and Physiology - Part B, 151, 278–285.

    Article  Google Scholar 

  • Padmini, E., & Vijaya Geetha, B. (2007a). A comparative seasonal pollution assessment study on estuary with respect to metal accumulation in Mugil cephalus. Oceanological and Hydrobiological Studies, 35, 1–13.

    Google Scholar 

  • Padmini, E., & Vijaya Geetha, B. (2007b). Seasonal influences on water quality parameters and pollution status of the Ennore estuary, Tamilnadu, India. Journal of Environmental Hydrology, 15, 1–9.

    Google Scholar 

  • Pascussi, J. M., Gerbal-Chaloin, S., Duret, C., Daujat-Chavanieu, M., Vilarem, M. J., & Maurel, P. (2008). The tangle of nuclear receptors that controls xenobiotic metabolism and transport: crosstalk and consequences. Annual Review of Pharmacology, 48, 1–32.

    Article  CAS  Google Scholar 

  • Raghunathan, M. B., & Srinivasan, M. (1983). Zooplankton dynamics and hydrographic features of Ennore estuary, Madras. Records of the Zoological Survey of India, 40, 1–30.

    Google Scholar 

  • Rahman, M. S., & Thomas, P. (2012). Effects of hypoxia exposure on hepatic cytochrome P450 1A (CYP1A) expression in Atlantic croaker: molecular mechanisms of CYP1A down-regulation. PloS One, 7(7), e40825. doi:10.1371/journal.pone.0040825.

    Article  CAS  Google Scholar 

  • Ryter, S. W., Otterbein, L. E., Morse, D., & Choi, A. M. (2002). Heme oxygenase/carbon monoxide signaling pathways: regulation and functional significance. Molecular and Cellular Biochemistry, 234(235), 249–263.

    Article  Google Scholar 

  • Schlenk, D., & Di-giulio, R. T. (2002). Biochemical responses as indicators of aquatic ecosystem health. In S. M. Adams (Ed.), Biological indicators of aquatic ecosystem stress (pp. 14–17). Bethesda: AFS.

    Google Scholar 

  • Schlenk, D., Celander, M., Gallagher, E. P., George, S., James, M., Kullman, S. W., van den Hurk, P., & Willett, K. (2008). Biotransformation in fishes. In R. T. Di Giulio & D. E. Hinton (Eds.), The toxicology of fishes (pp. 153–234). Boca Raton: CRC Press.

    Chapter  Google Scholar 

  • Semenza, G. L. (1998). Hypoxia-inducible factor 1: master regulator of O2 homeostasis. Current Opinion in Genetics and Development, 8, 588–594.

    Article  CAS  Google Scholar 

  • Semenza, G. L. (2005). Involvement of hypoxia-inducible factor 1 in pulmonary pathophysiology. Chest, 128, 592S–594S.

    Article  CAS  Google Scholar 

  • Shugart, L. R. (2000). DNA damage as a biomarker of exposure. Ecotoxicology, 9, 329–340.

    Article  CAS  Google Scholar 

  • Sies, H. (1986). Biochemistry of oxidative stress. Angewandte Chemie International Edition in English, 25, 1058–1071.

    Article  Google Scholar 

  • Slebos, D. J., Ryter, S. W., & Choi, A. M. K. (2003). Heme oxygenase-1 and carbon monoxide in pulmonary medicine. Respiratory Research, 4(1), 7.

    Article  Google Scholar 

  • Stegeman, J. J., & Hahn, M. E. (1994). Biochemistry and molecular biology of monooxygenases: current perspectives on forms, functions, and regulation of mechanisms of CYP1A regulation by hypoxia in fish cytochrome P450 in aquatic species. In D. C. Malins & G. K. Ostrander (Eds.), Aquatic toxicology (pp. 87–203). Boca Raton: Lewis Publishers.

    Google Scholar 

  • Strober, W. (2001). Trypan blue exclusion test of cell viability. Current Protocols in Immunology May, Appendix 3, Appendix 3B 18432654 (P, S, E, B, D).

  • Struznka, L., Chalimoniuk, M., & Sulkowski, G. (2005). The role of astroglia in Pb- exposed adult rat brain with respect to glutamate toxicity. Toxicology, 212, 185–194.

    Article  Google Scholar 

  • Tian, Y., Ke, S., Denison, M. S., Rabson, A. B., & Gallo, M. A. (1999). Ah receptor and NF-kappaB interactions, a potential mechanism for dioxin toxicity. Journal of Biological Chemistry, 274, 510–515.

    Article  CAS  Google Scholar 

  • Towbin, H., Staehelin, T., & Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America, 76, 4350–4354.

    Article  CAS  Google Scholar 

  • Van der Oost, R., Beyer, J., & Vermeulen, N. P. E. (2003). Fish bioaccumulation and biomarkers in environmental risk assessment: a review. Environmental Toxicology and Pharmacology, 13, 57–149.

    Article  Google Scholar 

  • Viarengo, A., Lowe, D., Bolognesi, C., Fabbri, E., & Koehler, A. (2007). The use of biomarkers in biomonitoring: a 2-tier approach assessing the level of pollutant-induced stress syndrome in sentinel organisms. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 146, 281–300.

    CAS  Google Scholar 

  • Vutkuru, S. (2005). Acute effects of Hexavalent chromium on survival, oxygen consumption, hematological parameters and some biochemical profiles of the Indian major carp, Labeo rohita. International Journal of Environmental Research and Public Health, 2, 456–462.

    Article  Google Scholar 

  • Wang, G. L., Jiang, B. H., Rue, E. A., & Semenza, G. L. (1995). Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proceedings of the National Academy of Sciences of the United States of America, 92, 5510–5514.

    Article  CAS  Google Scholar 

  • Whyte, J. J., Jung, R. E., Schmitt, C. J., & Tillitt, D. E. (2000). Ethoxyresorufin-O-deethylase (EROD) activity in fish as a biomarker of chemical exposure. Critical Reviews in Toxicology, 30, 347–570.

    Article  CAS  Google Scholar 

  • Xiao, H., Gu, Z., Wang, G., & Zhao, T. (2013). The possible mechanisms underlying the impairment of HIF-1α pathway signaling in hyperglycemia and the beneficial effects of certain therapies. International Journal of Medical Sciences, 10(10), 1412–1421.

    Article  Google Scholar 

  • Yang, X. M., Wang, Y. S., Zhang, J., Li, Y., Xu, J. F., Zhu, J., Zhao, W., Chu, D. K., & Wiedemann, P. (2009). Role of PI3K/Akt and MEK/ERK in mediating hypoxia-induced expression of HIF-1alpha and VEGF in laser-induced rat choroidal neovascularization. Investigative Ophthalmology & Visual Science, 50, 1873–1879.

    Article  Google Scholar 

Download references

Acknowledgments

The project was funded by University Grants Commission, New Delhi, India. Project referral number-UGC: 41-1281/2012 (SR) is acknowledged.

Conflict of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Author information

Authors and Affiliations

  1. P.G. Department of Biochemistry, Bharathi Women’s College, Affiliated to University of Madras, Chennai, 600108, Tamil Nadu, India

    Ekambaram Padmini & Jayachandran Tharani

Authors
  1. Ekambaram Padmini
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jayachandran Tharani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ekambaram Padmini.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Padmini, E., Tharani, J. Differential expression of HO-1 and CYP1A2 during up-regulation of ERK in stressed fish hepatocytes. Environ Monit Assess 187, 4147 (2015). https://doi.org/10.1007/s10661-014-4147-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10661-014-4147-1

Keywords

Search

Navigation