The effect of coadministration of α-tocopherol and ascorbic acid on arsenic trioxide-induced testicular toxicity in adult rats

Prabir Kumar Mukhopadhyay; Avijit Dey; Sanjit Mukherjee; Nirmal Kumar Pradhan

doi:10.1515/jbcpp-2012-0039

Article

The effect of coadministration of α-tocopherol and ascorbic acid on arsenic trioxide-induced testicular toxicity in adult rats

Prabir Kumar Mukhopadhyay , Avijit Dey , Sanjit Mukherjee and Nirmal Kumar Pradhan

Published/Copyright: August 15, 2013

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From the journal Journal of Basic and Clinical Physiology and Pharmacology Volume 24 Issue 4

Abstract

Background: Arsenic, acting as an endocrine disruptor, causes reproductive malfunctions. Studies have been undertaken to find out whether the co-supplementation of α-tocopherol and ascorbic acid (AT-AA) could reduce the arsenic-induced testicular toxicity caused by oxidative stress and resulting DNA damage.

Methods: Adult male Wistar rats (120±10 g) were given arsenic trioxide [3 mg/kg body weight (b.wt.) per day] for 30 consecutive days and the supplement group received α-tocopherol (400 mg/kg b.wt. per day) and ascorbic acid (200 mg/kg b.wt. per day). Reproductive functions were evaluated with respect to the histoarchitecture, gametokinetic activity, androgenic potential, glutathione-dependent antioxidant status and DNA damage of the testis.

Results: Arsenic treatment caused marked reduction in the relative weight of the testis (p<0.05) but showed no effect on body weight. The number of germ cells at stage VII of the spermatogenic cycle (p<0.01), the seminiferous tubular diameter (p<0.001) and Leydig cell nuclear area (p<0.01) were significantly reduced. Notable decrease in the activities of testicular Δ⁵, 3β-HSD (p<0.05) and 17β-HSD (p<0.01) with a concomitant fall in serum testosterone level (p<0.01) along with significant diminution in testicular glutathione S-transferase (p<0.05) activity and reduced glutathione level (p<0.01) were observed. Significant DNA damage (p<0.001) in spermatogenic cells was also noted. All these alterations including DNA strand breakage were seen to be protected with the coadministration of AT-AA.

Conclusions: The data suggest that the protection of testicular toxicity in arsenic-exposed adult rats is possible with combined coadministration of AT-AA.

Keywords: α-tocopherol and ascorbic acid; arsenic; genotoxic; oxidative stress; testis

Corresponding author: Dr. Prabir Kumar Mukhopadhyay, Associate Professor, Department of Physiology, Presidency University, 86/1 College Street, Kolkata-700073, India, Phone: +91-9433160257, E-mail: p.mukherjeepresi@gmail.com

The authors gratefully acknowledge the financial assistance from the Minor Research Project [No. F.PSW-074/09-10 (ERO)] provided by the University Grants Commission, New Delhi, India.

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article. Research financing played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

References

1. International Agency for Research on Cancer. In: IARC monographs on the evaluation of carcinogenic risk to humans – overall evaluations of carcinogenicity. An updating of IARC Monographs, Vols. 1 to 42 (Supplement 7). Lyon: International Agency for Research on Cancer, 1987:100.Search in Google Scholar

2. WHO. Environmental health criteria: arsenic. Geneva, Switzerland: WHO, 1981.Search in Google Scholar

3. Prozialeck WC, Edwards JR, Nebert DW, Woods JM, Barchowsky A, Atchison WD, et al. The vascular system as a target of metal toxicity. Toxicol Sci 2008;102:207–18.10.1093/toxsci/kfm263Search in Google Scholar

4. Chakraborti D, Rahman MM, Paul K, Chowdhury UK, Sengupta MK, Lodh D, et al. Arsenic calamity in the Indian subcontinent: what lessons have been learned? Talanta 2002;58:3–22.10.1016/S0039-9140(02)00270-9Search in Google Scholar

5. Waalkes MP, Ward JM, Liu J, Diwan BA. Transplacental carcinogenicity of inorganic arsenic in the drinking water, induction of hepatic, ovarian, pulmonary and adrenal tumors in mice. Toxicol Appl Pharmacol 2003;186:7–17.10.1016/S0041-008X(02)00022-4Search in Google Scholar

6. Mazumder DN, Guha A, Chakraborty AK, Ghosh A, Gupta JD, Chakraborty DP, et al. Chronic arsenic toxicity from drinking tube-well water in rural West Bengal. Bull WHO 1988;66:499–506.Search in Google Scholar

7. Sarkar M, Chaudhuri GR, Chattopadhyay A, Biswas NM. Effect of sodium arsenite on spermatogenesis, plasma gonadotrophins and testosterone in rats. Asian J Androl 2003;5:27–31.Search in Google Scholar

8. Sukla JP, Pandey K. Impaired spermatogenesis in arsenic-treated fresh water fish Colisa fasciatus (Bl and Sch). Toxicol Lett 1984;2:191–5.Search in Google Scholar

9. Hei T K, Filipic M. Role of oxidative damage in the genotoxicity of arsenic. Free Radic Biol Med 2004;37:574–81.10.1016/j.freeradbiomed.2004.02.003Search in Google Scholar

10. Kadirvel R, Sundaram K, Mani S, Samuel S, Elango N, Panneerselvam C, et al. Supplementation of ascorbic acid and α-tocopherol prevents arsenic-induced protein oxidation and DNA damage induced by arsenic in rats. Hum Exp Toxicol 2007;26:939–46.10.1177/0960327107087909Search in Google Scholar

11. Li X, Cobb CE, Hill KE, Burk RF, May JM. Mitochondrial uptake and recycling of ascorbic acid. Arch Biochem Biophys 2001;387:147–53.10.1006/abbi.2000.2245Search in Google Scholar

12. Samokyszyn VM, Miller DM, Reif DW, Aust SD. Iron-catalyzed lipid peroxidation. In: Vigo-Pelfrey C, editor. Membrane lipid oxidation, Vol. 1. Boca Raton, FL: CRC Press, 1990:101–27.Search in Google Scholar

13. Kini RD, Tripathi Y, Raghuveer CV, Pai SR, Ramswamy C, Nayanatara AK, et al. Protective role of vitamin E against cadmium chloride induced testicular damage in rats. J Physiol Biomed Sci 2009;22:12–6.Search in Google Scholar

14. Kini RD, Tripathi Y, Raghuveer CV, Pai SA, Ramaswamy C. Role of vitamin C as an antioxidant in cadmium chloride induced testicular damage. Int J Appl Biol Pharmacol Technol 2011;2:484–8.Search in Google Scholar

15. Kini RD, Tripathi Y, Raghuveer CV, Pai SR, Ramaswamy C. Combined effects of vitamin E and vitamin C supplementation on cadmium induced testicular morphology and oxidative stress. Res J Pharm Biol Chem Sci 2011;2:27–34.Search in Google Scholar

16. Samir AE, Bashandy. Beneficial effect of combined administration of vitamin C and vitamin E in amelioration of chronic lead hepatotoxicity. Egypt J Hosp Med 2006;23:371–84.10.21608/ejhm.2006.17948Search in Google Scholar

17. Momeni HR, Oryan S, Eskandari N. Effect of vitamin E on sperm number and testis histopathology of sodium arsenite-treated rats. Reprod Biol 2012;12:171–81.10.1016/S1642-431X(12)60084-9Search in Google Scholar

18. Sarkar S, Hazra J, Upadhyay SN, Singh RK, Roy Chowdhury A. Arsenic induced toxicity on testicular tissue of mice. Indian J Physiol Pharmacol 2008;52:84–90.Search in Google Scholar

19. Balakumar BS, Kumaresan S, Venugopal R, Sivasubramanian V. Clastogenic effect of sodium arsenite in experimental rats and ameliorative effects of antioxidant vitamins C and E. Indian J Sci Technol 2010;3:642–7.10.17485/ijst/2010/v3i6.22Search in Google Scholar

20. Dey A, Bose A, Mukhopadhyay PK. Testicular toxicity in arsenic exposed albino rats: ameliorative effects of ascorbic acid and α-tocopherol. Biomedicine 2011;31:53–63.Search in Google Scholar

21. Chappell WR, Beck BD, Brown KG, Chaney R, Cothern R, Cothern CR, et al. Inorganic arsenic: a need and an opportunity to improve risk assessment. Environ Health Perspect 1997;105:1060–7.10.1289/ehp.971051060Search in Google Scholar PubMed PubMed Central

22. Mukherjee S, Das D, Darbar D, Mitra, C. Dietary intervention affects arsenic generated nitric oxide and reactive intermediate toxicity in islet cell of rats. Curr Sci 2003;85:786–93.Search in Google Scholar

23. Leblond CP, Clermont. Definition of the stages of the cycle of the seminiferous epithelium in the rat. Ann N Y Acad Sci 1952;55:548–73.10.1111/j.1749-6632.1952.tb26576.xSearch in Google Scholar

24. Abercrombie, M. Estimation of nuclear population from microtome sections. Anat Res 1946;94:239–47.10.1002/ar.1090940210Search in Google Scholar

25. Clemont Y, Morgentaler H. Quantitative study of spermatogenesis in hypophysectomized rat. Endocrinology 1955;57:369–82.10.1210/endo-57-3-369Search in Google Scholar

26. Talaly P. Hydroxysteroid dehydrogenases. In: Clayton RB, editor. Methods in enzymology, Vol. V. New York: Academic Press, 1962:512–6.Search in Google Scholar

27. Jarabak GA. Soluble 17β-hydroxisteroid dehydrogenase on human placenta. In: Clayton RB, editor. Methods in enzymology, Vol. 1. New York: Academic Press, 1969:146–82.Search in Google Scholar

28. Zakowski JJ, Tsppel AL. Semi-automated system for measurement of glutathione in the assay of glutathione peroxidase. Anal Biochem 1978;89:430–6.10.1016/0003-2697(78)90372-XSearch in Google Scholar

29. Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 1974;249:713–39.Search in Google Scholar

30. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265–75.10.1016/S0021-9258(19)52451-6Search in Google Scholar

31. Singh NP. A rapid method for the preparation of single-cell suspensions from solid tissues. Cytometry 1998;31:229–32.10.1002/(SICI)1097-0320(19980301)31:3<229::AID-CYTO10>3.0.CO;2-TSearch in Google Scholar

32. Chowdhury AK. Dependence of testicular germ cells on hormone: a quantitative study in hypophysectomized testosterone treated rats. J Endocrinol 1979;82:33–40.10.1677/joe.0.0820331Search in Google Scholar

33. Murono EP, Payne AH. Testicular maturation in the rat: in vivo effect of gonadotropins and steroidogenic enzymes in hypophysectomized immature rats. Biol Reprod 1979;20:911–7.10.1095/biolreprod20.4.911Search in Google Scholar

34. Jana K, Jana S, Samanta PK. Effects of chronic exposure to sodium arsenite on hypothalamo-pituitary-testicular activities in adult rats: possible an estrogenic mode of action. Reprod Biol Endocrinol 2006;4:1–13.10.1186/1477-7827-4-9Search in Google Scholar

35. Barnes MM, Smith AJ. The effects of vitamin E deficiency on some enzymes of steroid biosynthesis. Int J Vitam Nutr Res 1975;45:396–403.Search in Google Scholar

36. Das P, Chowdhury M. Vitamin E-deficiency induced changes in ovary and uterus. Mol Cell Biochem 1999;198:151–6.10.1023/A:1006954032164Search in Google Scholar

37. Sharma MK, Buettner GR. Interaction of vitamin C and vitamin E during free radical stress in plasma: an ESR study. Free Radic Biol Med 1993;14:649–53.10.1016/0891-5849(93)90146-LSearch in Google Scholar

38. Chainy GB, Samantaray S, Samanta L. Testosterone-induced changes in testicular antioxidant system. Andrologia 1997;29:343–9.10.1111/j.1439-0272.1997.tb00328.xSearch in Google Scholar PubMed

39. Wang TC, Jan KY, Wang AS, Gurr JR. Trivalent arsenicals induce lipid peroxidation, protein carbonylation and oxidative DNA damage in human urothelial cells. Mutat Res 2006;615:75–86.10.1016/j.mrfmmm.2006.10.003Search in Google Scholar PubMed

40. Gerster H. Carotene, vitamin E and vitamin C in different stages of experimental carcinogenesis. Eur J Clin Nutr 1995;49:155–68.Search in Google Scholar

41. Dawson JR, Vahakangas K, Jernstrom B, Moldeus P. Glutathione conjugation by isolated lung cells and the isolated, perfused lung: effect of extracellular glutathione. Eur J Biochem 1984;138:439–43.10.1111/j.1432-1033.1984.tb07935.xSearch in Google Scholar PubMed

42. Scott N, Hatlelid KM, Mackinzie NE, Carter DE. Reactions of arsenic(III) and arsenic(V) species with glutathione. Chem Res Toxicol 1993;6:102–6.10.1021/tx00031a016Search in Google Scholar PubMed

43. Thirunavukkarasu C, Sakthisekaran D. Sodium selenite, dietary micronutrient, prevents the lymphocyte DNA damage induced by N-nitrosodiethylamine and phenobarbital promoted experimental hepatocarcinogenesis. J Cell Biochem 2003;88:578–88.10.1002/jcb.10362Search in Google Scholar PubMed

44. Nava-Hernandez MP, Hauad-Marroquín LA, Bassol-Mayagoitia S, García-Arenas G, Mercado-Hernández R, Echávarri-Guzmán MA, et al. Lead-, cadmium-, and arsenic-induced DNA damage in rat germinal cells. DNA Cell Biol 2009;28:241–8.10.1089/dna.2009.0860Search in Google Scholar PubMed

45. Momeni HR, Eskandari N. Effect of vitamin E on sperm parameters and DNA integrity in sodium arsenite-treated rats. Iran J Reprod Med 2012;10:249–56.Search in Google Scholar

46. Yamauchi H, Aminaka Y, Yoshida K, Sun GJ, Waalkes MP. Evaluation of DNA damage in patients with arsenic poisoning: urinary 8-hydrodeoxyguanine. Toxicol Appl Pharmacol 2004;198:291–6.10.1016/j.taap.2003.10.021Search in Google Scholar PubMed

47. Balakumar BS, Ramanathan K, Kumaresan S, Suresh R. DNA damage by sodium arsenite in experimental rats: ameliorative effects of antioxidant vitamins C and E. Indian J Sci Technol 2010;3:322–7.10.17485/ijst/2010/v3i3.12Search in Google Scholar

48. Valko M, Izakovic M, Mazur M, Rhodes CJ, Telser J. Role of oxygen radicals in DNA damage and cancer incidence. Mol Cell Biochem 2004;266:37–56.10.1023/B:MCBI.0000049134.69131.89Search in Google Scholar

49. Ramanathan K, Balakumar BS, Panneercellvam C. Effects of ascorbic acid and α-tocopherol on arsenic-induced oxidative stress. Hum Exp Toxicol 2002;21:675–80.10.1191/0960327102ht307oaSearch in Google Scholar PubMed

Received: 2012-8-10

Accepted: 2013-7-17

Published Online: 2013-08-15

Published in Print: 2013-11-01

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https://doi.org/10.1515/jbcpp-2012-0039

Keywords for this article

α-tocopherol and ascorbic acid; arsenic; genotoxic; oxidative stress; testis