Rat Testicular Mitochondrial Antioxidant Defence System and its Modulation By Aging

Abstract

Accumulation of oxidative damage caused by reactive oxygen species (ROS) underlies fundamental changes found during aging. In the present study, age related effect on testicular mitochondrial oxidant generation and antioxidant defence profile was investigated in Wistar rats at 3 months (young adults), 12 months (old adults) and 24 months (senescent animals) of age. Mitochondrial oxidative stress parameters viz., lipid peroxidation (LPx), protein carbonylation, hydrogen peroxide (H202) generation and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), levels of total, oxidized (GSSG) and reduced glutathione (GSH) were studied to find out their roles in maintenance of mitochondrial glutathione redox pool as a function of age. Increased levels of LPx, H202 and decreased GSH content accompanied by a decline in activities of SOD, GPx and GR with advancing age suggest that antioxidant defense profile of testicular mitochondria exhibit age related alterations which might play a critical role in regulating physiological functions of the testis such as steroidogenesis and spermatogenesis.

References

  1. 1.

    Cao, L., Leers-Sucheta, S., Azhar, S. (2004) Aging alters the functional expression of enzymatic and non-enzymatic antioxidant defence system in testicular rat Leydig cells. J. Steroid Biochem. Mol. Biol. 88, 61–70.

    CAS  Article  Google Scholar 

  2. 2.

    Capel, F., Rimbert, V., Lioger, D., Diot, A., Rousset, P., Mirand, P. P., Boirie, Y., Morio, B., Mosoni, L. (2005) Due to reverse electron transfer, mitochondrial H2O2 release increases with age in human vastus lateralis muscle although oxidative capacity is preserved. Mech. Ageing Dev. 126, 505–511.

    CAS  Article  Google Scholar 

  3. 3.

    Carmagnol, F., Sinet, P. M., Jerome, H. (1983) Selenium-dependent and non-selenium-dependent glutathione peroxidases in human tissue extracts. Biochim. Biophys. Acta 759, 49–57.

    CAS  Article  Google Scholar 

  4. 4.

    Chen, J., Irizarry, R. A., Luo, L., Zirkin, B. R. (2004) Leydig cells gene expression: Effect of age and caloric restriction. Expt. Gerontol. 39, 31–40.

    CAS  Article  Google Scholar 

  5. 5.

    Cohen, G., Dembiec, D., Marcus, J. (1970) Measurement of catalase activity in tissue extracts. Anal. Biochem. 34, 30–38.

    CAS  Article  Google Scholar 

  6. 6.

    Das, K., Samanta, L., Chainy, G. B. N. (2000) A modified spectrophotometric assay of superoxide dismutase using nitrite formation of superoxide radicals. Ind. J. Biochem. Biophys. 37, 201–204.

    CAS  Google Scholar 

  7. 7.

    de la Asuncion, J. G., Millan, A., Pla, R., Bruseghini, L., Esteras, A., Pallardo, F. V., Sastre, J., Vina, J. (1996) Mitochondrial glutathione oxidation correlates with age-associated oxidative damage to mitochondrial DNA. FASEB J. 10, 333–338.

    Article  Google Scholar 

  8. 8.

    Fucci, L., Oliver, C. N., Coon, M. J., Stadtman, E. R. (1983) Inactivation of key metabolic enzymes by mixed function oxidation reactions: possible implication in protein turnover and aging. Proc. Nat. Acad. Sci. USA 80, 1521–1525.

    CAS  Article  Google Scholar 

  9. 9.

    Harman, D. (1956) Aging: a theory based on free radical and radiation chemistry. J. Gerontol. 11, 298–300.

    CAS  Article  Google Scholar 

  10. 10.

    Kaur, A. (2005) Changes in peroxidase activity and structure of testes, epididymis and seminal vesicles in aging Swiss albino mice. Ind. J. Gerontol. 19, 119–126.

    Google Scholar 

  11. 11.

    Kumaran, S., Deepak, B., Naveen, B., Panneerselvam, C. (2003) Effects of levocarnitine on mitochondrial antioxidant systems and oxidative stress in aged rats. Drugs R.D. 4, 141–147.

    CAS  Article  Google Scholar 

  12. 12.

    Lee, C. Y. (1982) Glucose-6-phosphate dehydrogenase from mouse. Methods Enzymol. 89, 252–260.

    CAS  Article  Google Scholar 

  13. 13.

    Lee, H. C., Wei, Y. H. (2000) Mitochondrial role in life and death of the cell. J. Biomed. Sci. 7, 2–15.

    CAS  Article  Google Scholar 

  14. 14.

    Levine, R. L., Garland, D., Oliver, C. N., Amici, A., Climent, I., Lenz, A. G., Ahn, B. W., Shaltiel, S., Stadtman, E. R. (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol. 186, 464–478.

    CAS  Article  Google Scholar 

  15. 15.

    Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. (1951) Protein measurement with the Folin-phenol reagent. J. Biol. Chem. 193, 265–275.

    CAS  Google Scholar 

  16. 16.

    Luo, L., Chen, H., Trush, M. A., Show, M. D., Anway, M. D., Zirkin, B. R. (2006) Aging and the brown Norway rat Leydig cell antioxidant defence system. J. Androl. 27, 240–247.

    CAS  Article  Google Scholar 

  17. 17.

    Massey, V., Williams, C. H. (1965) On the reaction mechanism of yeast glutathione reductase. J. Biol. Chem. 240, 4470–4481.

    CAS  PubMed  Google Scholar 

  18. 18.

    Matsubara, L. S., Machado, P. E. A. (1991) Age related changes of glutathione content, glutathione reductase and glutathione peroxidase activity of human erythrocytes. Brazil. J. Med. Biol. Res. 24, 449–459.

    CAS  Google Scholar 

  19. 19.

    Miquel, J., Economos, A. C, Fleming, J., Johnson, J. E. Jr. (1980) Mitochondrial role in cell aging. Exp. Gerontol. 15, 579–591.

    Article  Google Scholar 

  20. 20.

    Nagley, P., Wei, Y. H. (1998) Ageing and mammalian mitochondrial genetics. Trends Genet. 14, 513–517.

    CAS  Article  Google Scholar 

  21. 21.

    Navarro, A., Boveris, A. (2004) Rat brain and liver mitochondria develop oxidative stress and lose enzymatic activities on aging. Am. J. Physiol. Regul. Integr. Comp. Physiol. 287, R1244–R1249.

    CAS  Article  Google Scholar 

  22. 22.

    Ohkawa, H., Ohishi, N., Yagi, K. (1979) Assay for lipid peroxides in animal tissue by thiobarbituric acid reaction. Anal. Biochem. 95, 351–358.

    CAS  Article  Google Scholar 

  23. 23.

    Paglia, D. E., Valentine, W. N. (1967) Studies on quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J. Lab. Clin. Med. 70, 158–169.

    CAS  PubMed  Google Scholar 

  24. 24.

    Peltola, V., Huhtaniemi, I., Ahotupa, M. (1992) Antioxidant enzyme activity in the maturing rat testis. J. Androl. 13, 450–455.

    CAS  PubMed  Google Scholar 

  25. 25.

    Pick, E., Keisari, Y. (1981) Superoxide anion and hydrogen peroxide production by chemically elicited peritoneal macrophages-induction by multiple nonphagocytic stimuli. Cell. Immunol. 59, 301–318.

    CAS  Article  Google Scholar 

  26. 26.

    Sahoo, D. K., Roy, A., Bhanja, S., Chainy, G. B. N. (2005) Experimental hyperthyroidism-induced oxidative stress and impairment of antioxidant defence system in rat testis. Ind. J. Exp. Biol. 43, 1058–1067.

    CAS  Google Scholar 

  27. 27.

    Sahoo, D. K., Roy, A., Bhanja, S., Chainy, G. B. N. (2008) Hypothyroidism impairs antioxidant defence system and testicular physiology during development and maturation. Gen. Comp. Endocrinol. 156, 63–70.

    CAS  Article  Google Scholar 

  28. 28.

    Sahoo, D. K., Roy, A., Chattopadhyay, S., Chainy, G. B. N. (2007) Effect of T3 treatment on glutathione redox pool and its metabolizing enzymes in mitochondrial and post-mitochondrial fractions of adult rat testes. Ind. J. Exp. Biol. 45, 338–346.

    CAS  Google Scholar 

  29. 29.

    Samanta, L., Chainy, G. B. N. (1997) Comparison of hexachlorocyclohexane-induced oxidative stress in the testis of immature and adult rats. Comp. Biochem. Physiol. 118, 319–327.

    CAS  Google Scholar 

  30. 30.

    Samanta, L., Chainy, G. B. N. (2002) Effect of age on testicular antioxidant defence system. Ind. J. Gerontol. 16, 69–78.

    Google Scholar 

  31. 31.

    Samanta, L., Roy, A., Chainy, G. B. N. (1999) Changes in rat testicular antioxidant defence profile as a function of age and its impairment by hexachlorocyclohexane during critical stages of maturation. Andrologia 31, 83–90.

    CAS  Article  Google Scholar 

  32. 32.

    Sastre, J., Pallardo, F. V., Vina, J. (2003) The role of mitochondrial oxidative stress in aging. Free Rad. Biol. Med. 35, 1–8.

    CAS  Article  Google Scholar 

  33. 33.

    Sharma, R., Nakamura, A., Takahashi, R., Nakamoto, H., Goto, S. (2006) Carbonyl modification in rat liver histones: Decrease with age and increase by dietary restriction. Free Rad. Biol. Med. 40, 1179–1184.

    CAS  Article  Google Scholar 

  34. 34.

    Shigenaga, M. K., Hagen, T. M., Ames, B. N. (1994) Oxidative damage and mitochondrial decay in aging. Proc. Natl. Acad. Sci. USA 91, 10771–10778.

    CAS  Article  Google Scholar 

  35. 35.

    Sohal, R. S. (1993) The free radical hypothesis of aging: an appraisal of the current status. Aging Clin. Exp. Res. 5, 3–17.

    CAS  Article  Google Scholar 

  36. 36.

    Sohal, R. S., Arnold, L. A., Sohal, B. H. (1990) Age related changes in antioxidant enzymes and prooxidant generation in tissues of the rat with special reference to parameters in two insect species. Free Radic. Biol. Med. 10, 495–500.

    Article  Google Scholar 

  37. 37.

    Teaf, C. M., Harbison, R. D., Bishop, J. B. (1985) Germ-cell mutagenesis and GSH depression in reproductive tissue of the F-344 rat induced by ethyl methanesulfonate. Mut. Res. 144, 93–98.

    CAS  Article  Google Scholar 

  38. 38.

    Zirkin, B. R., Chen, H. (2000) Regulation of leydig cell steroidogenic function during aging. Biol. Reprod. 63, 977–981.

    CAS  Article  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to G. B. N. Chainy.

Rights and permissions

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Cite this article

Sahoo, D.K., Roy, A. & Chainy, G.B.N. Rat Testicular Mitochondrial Antioxidant Defence System and its Modulation By Aging. BIOLOGIA FUTURA 59, 413–424 (2008). https://doi.org/10.1556/ABiol.59.2008.4.3

Download citation

Keywords

  • Aging
  • rat
  • testes
  • mitochondria
  • reactive oxygen species
  • oxidative stress