Skip to main content

Advertisement

SpringerLink
Log in

Hydrogen peroxide leads to cell damage and apoptosis in the gill of the freshwater crab Sinopotamon henanense (Crustacea, Decapoda)

  • CHALLENGES IN AQUATIC SCIENCES
  • Published:
Hydrobiologia Aims and scope Submit manuscript

Abstract

Hydrogen peroxide (H2O2), a major reactive oxygen species, has been shown to be a critical mediator of apoptosis induced by several toxic metals such as cadmium. In this study, we used the freshwater crab Sinopotamon henanense to study whether H2O2 can cause apoptosis in gill cells. The crabs were incubated in H2O2 and the DNA fragmentation, ultrastructural changes and caspase-3/8/9 activities were measured. The results showed that in freshwater crab, H2O2 was found to induce apoptosis, as confirmed by DNA fragmentation analysis and morphological observation of transmission electron microscopy. This apoptosis occurs in a concentration-dependent pattern. During the apoptotic process, caspase-3, caspase-8 and caspase-9 were activated by H2O2. In addition, multiple physiological and pathological changes of gill cells were discovered after 24 h exposure to 5 mM H2O2, including aggregation and condensation of nuclear chromatin, appearance of extremely irregular nuclei with finger-like buds, disappearance of the organelles around the nuclei, swollen and dissolved cristae of mitochondria. We propose that H2O2-induced stress leads to mitochondria lesions oxidative injury and triggers apoptotic response through the caspase pathway in freshwater crab.

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

Similar content being viewed by others

References

  • Anuradha, C. D., S. Kanno & S. Hirano, 2001. Oxidative damage to mitochondria is a preliminary step to caspase-3 activation in fluoride-induced apoptosis in HL-60 cells. Free Radical Biology & Medicine 31: 367–373.

    Article  CAS  Google Scholar 

  • Avery, S. V., 2011. Molecular targets of oxidative stress. Biochemical Journal 434: 201–210.

    Article  PubMed  CAS  Google Scholar 

  • Barbouti, A., P. T. Doulias, L. Nousis, M. Tenopoulou & D. Galaris, 2002. DNA damage and apoptosis in hydrogen peroxide-exposed Jurkat cells: bolus addition versus continuous generation of H2O2. Free Radical Biology & Medicine 33: 691–702.

    Article  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  PubMed  CAS  Google Scholar 

  • Brennan, L. A., G. M. Morris, G. R. Wasson, B. M. Hannigan & Y. A. Barnett, 2000. The effect of vitamin C or vitamin E supplementation on basal and H2O2-induced DNA damage in human lymphocytes. British Journal of Nutrition 84: 195–202.

    Article  PubMed  CAS  Google Scholar 

  • Cabiscol, E., J. Tamarit & J. Ros, 2000. Oxidative stress in bacteria and protein damage by reactive oxygen species. International Microbiology 3: 3–8.

    PubMed  CAS  Google Scholar 

  • Caciari, T., A. Sancini, F. Tomei, L. Antetomaso, G. Tomei, B. Scala, F. Sinibaldi, C. Di Pastena, G. Andreozzi, N. Nardone, M. P. Schifano, P. Corbosiero, A. Capozzella, S. De Sio & M. Ciarrocca, 2012. Cadmium blood/urine levels and blood pressure in workers occupationally exposed to urban stressor. Annali du Igiene 24: 417–428.

    CAS  Google Scholar 

  • Cardone, M. H., N. Roy, H. R. Stennicke, G. S. Salvesen, T. F. Franke, E. Stanbridge, S. Frisch & J. C. Reed, 1998. Regulation of cell death protease caspase-9 by phosphorylation. Science 282: 1318–1321.

    Article  PubMed  CAS  Google Scholar 

  • Chen, J., X. Zhou, L. Feng, Y. Liu & J. Jiang, 2009a. Effects of glutamine on hydrogen peroxide-induced oxidative damage in intestinal epithelial cells of Jian carp (Cyprinus carpio var. Jian). Aquaculture 288: 285–289.

    Article  CAS  Google Scholar 

  • Chen, L., L. Liu, J. Yin, Y. Luo & S. Huang, 2009b. Hydrogen peroxide-induced neuronal apoptosis is associated with inhibition of protein phosphatase 2A and 5, leading to activation of MAPK pathway. The International Journal of Biochemistry & Cell Biology 41: 1284–1295.

    Article  CAS  Google Scholar 

  • Chen, Y. W., C. Y. Yang, C. F. Huang, D. Z. Hung, Y. M. Leung & S. H. Liu, 2009c. Heavy metals, islet function and diabetes development. Islets 1(3): 169–176.

    Article  PubMed  Google Scholar 

  • Cuypers, A., M. Plusquin, T. Remans, M. Jozefczak, E. Keunen, H. Gielen, K. Opdenakker, A. R. Nair, E. Munters, T. J. Artois, T. Nawrot, J. Vangronsveld & K. Smeets, 2010. Cadmium stress: an oxidative challenge. Biometals 23: 927–940.

    Article  PubMed  CAS  Google Scholar 

  • Dumont, A., S. P. Hehner, T. G. Hofmann, M. Ueffing, W. Dröge & M. L. Schmitz, 1999. Hydrogen peroxide-induced apoptosis is CD95-independent requires the release of mitochondria-derived reactive oxygen species and the activation of NF-κB. Oncogene 18: 747–757.

    Article  PubMed  CAS  Google Scholar 

  • El Golli-Bennour, E. & H. Bacha, 2011. Hsp70 expression as biomarkers of oxidative stress: mycotoxins’ exploration. Toxicology 287: 1–7.

    Article  PubMed  Google Scholar 

  • Enari, M., H. Sakahira, H. Yokoyama, K. Okawa, A. Iwamatsu & S. Nagata, 1998. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 28: 393–396.

    Google Scholar 

  • Facchinetti, F., S. Furegato, S. Terrazzino & A. Leon, 2002. H2O2 induces upregulation of Fas and Fas ligand expression in NGF-differentiated PC12 cells: modulation by cAMP. Journal of Neuroscience Research 69: 178–188.

    Article  PubMed  CAS  Google Scholar 

  • Feeney, L. & E. R. Berman, 1976. Oxygen toxicity: membrane damage by free radicals. Investigative Ophthalmology 15: 789–792.

    PubMed  CAS  Google Scholar 

  • Feng, B., W. L. Ye, L. J. Ma, Y. Fang, Y. A. Mei & S. M. Wei, 2012. Hydrogen peroxide enhanced Ca2+-activated BK currents and promoted cell injury in human dermal fibroblasts. Life Science 90: 424–431.

    Article  CAS  Google Scholar 

  • Franco, R., R. Sánchez-Olea, E. M. Reyes-Reyes & M. I. Panayiotidis, 2009. Environmental toxicity, oxidative stress and apoptosis: Ménage à Trois. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 674: 3–22.

    Article  CAS  Google Scholar 

  • Gautam, D. K., M. M. Misro, S. P. Chaki & N. Sehgal, 2006. H2O2 at physiological concentrations modulates Leydig cell function inducing oxidative stress and apoptosis. Apoptosis 11: 39–46.

    Article  PubMed  CAS  Google Scholar 

  • Gomes, P., S. Simão, V. Lemos, J. S. Amaral & P. Soares-da-Silva, 2013. Loss of oxidative stress tolerance in hypertension is linked to reduced catalase activity and increased c-Jun NH2-terminal kinase activation. Free Radical Biology & Medicine 56: 112–122.

    Article  CAS  Google Scholar 

  • Han, H., H. Wang, H. Long, S. Nattel & Z. Wang, 2001. Oxidative preconditioning and apoptosis in L-cells: roles of protein kinase B and mitogen-activated protein kinases. Journal of Biological Chemistry 276: 26357–26364.

    Article  PubMed  CAS  Google Scholar 

  • Issartel, J., V. Boulo, S. Wallon, O. Geffard & G. Charmantier, 2010. Cellular and molecular osmoregulatory responses to cadmium exposure in Gammarus fossarum (Crustacea, Amphipoda). Chemosphere 81: 701–710.

    Article  PubMed  CAS  Google Scholar 

  • Izawa, S., Y. Inoue & A. Kimura, 1995. Oxidative stress response in yeast: effect of glutathione on adaptation to hydrogen peroxide stress in Saccharomyces cerevisiae. FEBS Letter 368: 73–76.

    Article  CAS  Google Scholar 

  • Jacobson, M. D., 1996. Reactive oxygen species and programmed cell death. Trends in Biochemical Sciences 21: 83–86.

    Article  PubMed  CAS  Google Scholar 

  • Jänicke, R. U., M. L. Sprengart, M. R. Wati & A. G. Porter, 1998. Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. Journal of Biological Chemistry 273: 9357–9360.

    Article  PubMed  Google Scholar 

  • Konishi, H., M. Tanaka, Y. Takemura, H. Matsuzaki, Y. Ono, U. Kikkawa & Y. Nishizuka, 1997. Activation of protein kinase C by tyrosine phosphorylation in response to H2O2. Proceedings of the National Academy of Sciences 94: 11233–11237.

    Article  CAS  Google Scholar 

  • Kwon, D., C. Choi, J. Lee, K. O. Kim, J. D. Kim, S. J. Kim & I. H. Choi, 2001. Hydrogen peroxide triggers the expression of Fas/FasL in astrocytoma cell lines and augments apoptosis. Journal of Neuroimmunology 113: 1–9.

    Article  PubMed  CAS  Google Scholar 

  • Lee, R. F. & S. Steinert, 2003. Use of the single cell gel electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. Mutation Research 544: 43–64.

    Article  PubMed  CAS  Google Scholar 

  • Li, H. L., H. Zhu, C. J. Xu & J. Y. Yuan, 1998. Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 94: 491–501.

    Article  PubMed  CAS  Google Scholar 

  • Li, R., Y. Zhou, L. Wang & G. Ren, 2011. Low-molecular-weight-chitosan ameliorates cadmium-induced toxicity in the freshwater crab, Sinopotamon yangtsekiense. Ecotoxicology and Environmental Safety 74: 1164–1170.

    Article  PubMed  CAS  Google Scholar 

  • Liang, F. Q. & B. F. Godley, 2003. Oxidative stress-induced mitochondrial DNA damage in human retinal pigment epithelial cells: a possible mechanism for RPE aging and age-related macular degeneration. Experimental Eye Research 76: 397–403.

    Article  PubMed  CAS  Google Scholar 

  • Liang, D., L. Xiang, M. Yang, X. Zhang, B. Guo, Y. Chen, L. Yang & J. Cao, 2013. ZnT7 can protect MC3T3-E1 cells from oxidative stress-induced apoptosis via PI3K/Akt and MAPK/ERK signaling pathways. Cellular Signalling 25: 1126–1135.

    Article  PubMed  CAS  Google Scholar 

  • Liu, D., B. Yan, J. Yang, W. Lei & L. Wang, 2011. Mitochondrial pathway of apoptosis in the hepatopancreas of the freshwater crab Sinopotamon yangtsekiense exposed to cadmium. Aquatic Toxicology 105: 394–402.

    Article  PubMed  CAS  Google Scholar 

  • Lu, H., Z. Y. Zhu, L. L. Dong, X. M. Jia, X. R. Sun, L. Yan, Y. F. Chai, Y. Y. Jiang & Y. Y. Cao, 2011. Lack of trehalose accelerates H2O2-induced Candida albicans apoptosis through regulating Ca2+ signaling pathway and caspase activity. PLoS One 6: e15808.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Luo, X., B. Chen, R. Zheng, P. Lin, J. Li & H. Chen, 2010. Hydrogen peroxide induces apoptosis through the mitochondrial pathway in rat Schwann cells. Neuroscience Letters 485: 60–64.

    Article  PubMed  CAS  Google Scholar 

  • Martin, D., M. Salinas, N. Fujita, T. Tsuruo & A. Cuadrado, 2002. Ceramide and reactive oxygen species generated by H2O2 induce caspase-3-independent degradation of Akt/protein kinase B. Journal of Biological Chemistry 277: 42943–42952.

    Article  PubMed  CAS  Google Scholar 

  • Matsuzawa, A. & H. Ichijo, 2001. Molecular mechanisms of the decision between life and death: regulation of apoptosis by apoptosis signal-regulating kinase 1. Journal of Biochemistry 130: 1–8.

    Article  PubMed  CAS  Google Scholar 

  • Nawrot, T., M. Plusquin, J. Hogervorst, H. A. Roels, H. Celis, L. Thijs, J. Vangronsveld, E. Van Hecke & J. A. Staessen, 2006. Environmental exposure to cadmium and risk of cancer: a prospective population-based study. The Lancet Oncology 7: 119–126.

    Article  PubMed  CAS  Google Scholar 

  • Qin, Q., S. J. Qin, L. Wang & W. W. Lei, 2012. Immune responses and ultrastructural changes of hemocytes in freshwater crab Sinopotamon henanense exposed to elevated cadmium. Aquatic Toxicology 106–107: 140–146.

    Article  PubMed  Google Scholar 

  • Rhee, S. G., 1999. Redox signaling: hydrogen peroxide as intracellular messenger. Experimental and Molecular Medicine 31: 53–59.

    Article  PubMed  CAS  Google Scholar 

  • Saikumar, P., Z. Dong, V. Mikhailov, M. Denton, J. M. Weinberg & M. A. Venkatachalam, 1999. Apoptosis: definition, mechanisms, and relevance to disease. The American Journal of Medicine 107: 489–506.

    Article  PubMed  CAS  Google Scholar 

  • Salvesen, G. S. & V. M. Dixit, 1997. Caspases: intracellular signaling by proteolysis. Cell 91: 443–446.

    Article  PubMed  CAS  Google Scholar 

  • Sokolova, I. M., S. Evans & F. M. Hughes, 2004. Cadmium-induced apoptosis in oyster hemocytes involves disturbance of cellular energy balance but no mitochondrial permeability transition. Journal of Experimental Biology 207: 3369–3380.

    Article  PubMed  CAS  Google Scholar 

  • Stridh, H., M. Kimland, D. P. Jones, S. Orrenius & M. B. Hampto, 1998. Cytochrome c release and caspase activation in hydrogen peroxide- and tributyltin-induced apoptosis. FEBS Letters 429: 351–355.

    Article  PubMed  CAS  Google Scholar 

  • Suzuki, Y. J., H. J. Forman & A. Sevanian, 1997. Oxidants as stimulators of signal transduction. Free Radical Biology & Medicine 22: 269–285.

    Article  CAS  Google Scholar 

  • Suzuki, M., K. Aoshiba & A. Nagai, 2006. Oxidative stress increases Fas ligand expression in endothelial cells. Journal of Inflammation 19: 3–11.

    Google Scholar 

  • Taddei, F., V. Scarcelli, G. Frenzilli & M. Nigro, 2001. Genotoxic hazard of pollutants in cetaceans: DNA damage and repair evaluated in the bottlenose dolphin (Tursiops truncatus) by the Comet Assay. Marine Pollution Bulletin 42: 324–328.

    Article  PubMed  CAS  Google Scholar 

  • Takada, Y., A. Mukhopadhyay, G. C. Kundu, G. H. Mahabeleshwar, S. Singh & B. B. Aggarwal, 2003. Hydrogen peroxide activates NF-kappa B through tyrosine phosphorylation of I kappa B alpha and serine phosphorylation of p65: evidence for the involvement of I kappa B alpha kinase and Syk protein-tyrosine kinase. Journal of Biological Chemistry 278: 24233–24241.

    Article  PubMed  CAS  Google Scholar 

  • Tamagno, E., G. Robino, A. Obbili, P. Bardini, M. Aragno, M. Parola & O. Danni, 2003. H2O2 and 4-hydroxynonenal mediate amyloid β-induced neuronal apoptosis by activating JNKs and p38MAPK. Experimental Neurology 180: 144–155.

    Article  PubMed  CAS  Google Scholar 

  • Thévenod, F., 2009. Cadmium and cellular signaling cascades: to be or not to be? Toxicology and Applied Pharmacology 238: 221–239.

    Article  PubMed  Google Scholar 

  • Turner, N. A., F. Xia, G. Azhar, X. Zhang, L. Liu & J. Y. Wei, 1998. Oxidative stress induces DNA fragmentation and caspase activation via the c-Jun NH2-terminal kinase pathway in H9c2 cardiac muscle cells. Journal of Molecular and Cellular Cardiology 30: 1789–1801.

    Article  PubMed  CAS  Google Scholar 

  • Wang, J. X., P. P. Zhang, Q. Q. Shen, Q. Wang, D. M. Liu, J. Li & L. Wang, 2013. The effects of cadmium exposure on the oxidative state and cell death in the gill of freshwater crab Sinopotamon henanense. PLoS One 8: e64020.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Wyllie, A. H., 1980. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555–556.

    Article  PubMed  CAS  Google Scholar 

  • Yang, W. & S. T. Omaye, 2009. Air pollutants, oxidative stress and human health. Mutation Research 674: 45–54.

    Article  PubMed  CAS  Google Scholar 

  • Yu, Y., J. R. Du, C. Y. Wang & Z. M. Qin, 2008. Protection against hydrogen peroxide-induced injury by Z-ligustilide in PC12 cells. Experimental Brain Research 184: 307–312.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, L., H. Yu, Y. Sun, X. Lin, B. Chen, C. Tan, G. Cao & Z. Wang, 2007. Protective effects of salidroside on hydrogen peroxide-induced apoptosis in SH-SY5Y human neuroblastoma cells. European Journal of Pharmacology 564: 18–25.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, S., J. Ye & G. Dong, 2010. Neuroprotective effect of baicalein on hydrogen peroxide-mediated oxidative stress and mitochondrial dysfunction in PC12 cells. Journal of Molecular Neuroscience 40: 311–320.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to the National Natural Science Foundation of China (no. 31272319) and China Postdoctoral Science Foundation (no. 2012M511081) for financial support. We thank Prof. Enmin Zou for language editing of the manuscript.

Author information

Authors and Affiliations

  1. School of Life Science, Shanxi University, Taiyuan, 030006, China

    Jinxiang Wang, Qian Wang, Na Liu, Weixin Jing & Lan Wang

  2. School of Life Science, Shanxi Datong University, Datong, 037009, China

    Jinxiang Wang & Feng Zhou

Authors
  1. Jinxiang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qian Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Na Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Weixin Jing
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Feng Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lan Wang.

Additional information

Jinxiang Wang and Qian Wang contributed equally to the work.

Guest editors: Jiang-Shiou Hwang & Koen Martens / Challenges in Aquatic Sciences

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, J., Wang, Q., Liu, N. et al. Hydrogen peroxide leads to cell damage and apoptosis in the gill of the freshwater crab Sinopotamon henanense (Crustacea, Decapoda). Hydrobiologia 741, 13–21 (2014). https://doi.org/10.1007/s10750-013-1760-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10750-013-1760-x

Keywords

Search

Navigation