EFFECT OF ASCORBIC ACID AND ALPHA TOCOPHEROL SUPPLEMENTATION ON ACUTE RESTRAINT STRESS INDUCED CHANGES IN TESTOSTERONE, CORTICOSTERONE AND NOR EPINEPHRINE LEVELS IN MALE SPRAGUE DAWLEY RATS
AbstractBackground: Stress of various origins suppresses male reproductive functions through releasing stress hormones. Antioxidant like ascorbic acid (AA) and alpha tocopherol (AT) have been thought to protect the body against stress induced damage. Whether, these antioxidants confer protection against the stress induced increased levels of corticosterone and nor-epinephrine, and decreased testosterone secretion have been investigated in this study. Methods: This quasi experimental study was carried out at the Department of Physiology, Army Medical College Rawalpindi in collaboration with National Institute of Health, Islamabad during March to September 2009. Eighty male Sprague Dawley rats were divided into five groups with sixteen rats in each group. Group-I served as the control without stress while group-II was exposed to restraint stress for 6 hours, group-III was administered AA, group-IVAT and group-V was supplemented with both the antioxidants along with standard diet for one month. All antioxidant supplemented groups were exposed to restraint stress for 6 hours. Immediately after the stress episode, blood sample was obtained for the assay of serum testosterone, serum corticosterone by EIA and plasma nor-epinephrine levels by ELISA. Data were analyzed on SPSS-13 and p-value less than 0.05 was considered significant. Results: Acute restraint stress resulted in a statistically significant rise in corticosterone and nor-epinephrine levels and fall in serum testosterone levels. AA supplementation for one month revealed insignificant changes in stress induced hormonal parameters. AT alone and in combination with ascorbic acid prevented the fall in testosterone level as well as rise in corticosterone, however nor-epinephrine levels remained unchanged. Conclusion: Supplementation with AT alone or in combination with AA prevent reduction in testosterone and rise in corticosterone levels while keeping the nor-epinephrine levels unchanged after acute restraint stress in Sprague Dawley rats.Keywords: Acute restraint stress, testosterone, corticosterone, nor-epinephrine, ascorbic acid, alpha tocopherol
Chrousos GP, Gold PW. The concepts of stress and stress system disorders. Overview of physical and behavioural homeostasis. JAMA 1992;267:1244–52.
Lenze A, Lombardo F, Salacone P, Gandini L, Jannini EA. Stress, sexual dysfunctions and male infertility. J Endocrinol Invest 2003;26(3 Suppl):72–6.
McEwen BS. The neurobiology of stress: from serendipity to clinical relevance. Brain Res 2000;886(1–2):172–89.
Ozawa N, Goda N, Makino N, Yamaguchi T, Yoshimura Y, Suematsu M. Leydig cell derived hemeoxygenase –1 regulates apoptosis of premeiotic germ cells in response to stress. J Clin Invest 2002;109:547–67.
Fenster L, Katz DF, Wyrobek AJ, Pieper C, Rempel DM, Oman D, et al. Effects of psychological stress on human semen quality. J Androl 1997;18:194–202.
Bauer ME, Perks P, Lightman SL, Shanks N. Restraint stress is associated with changes in glucocorticoid immunoregulation. Physiol Behav 2001;73:525–32.
Cacioppo JT, Malarkey WB, Kiecolt-Glaser JK, Uchino BN, Sgoutas-Emch SA, Sheridan JF, et al. Heterogeneity in neuroendocrine and immune responses to brief psychological stressors as a function of autonomic cardiac activation. Psychosom Med 1995;57:154–64.
Zaidi SM, Al-Qirim TM, Hoda N, Banu N. Modulation of restraint stress induced oxidative changes in rats by antioxidant vitamins. J Nutr Biochem 2003;14:633–6.
Dong Q, Salva A, Sottas CM, Niu E, Holmes M, Hardy MP. Rapid glucocorticoid mediation of suppressed testosterone biosynthesis in male mice subjected to immobilization stress. J Androl 2004;25:973–81.
Hales DB, Payne AH. Glucocorticoid mediated repression of P450 scc mRNA and de novo synthesis in cultured Leydig cells. Endocrinology 1989;124:2099–104.
Akinbami MA, Taylor MF, Collins DE, Mann DR. Effect of peripheral and central acting opioid anagonist on the testicular response to stress in rats. Neuroendocrinology 1994;59:343–8.
Montoro J, Mullol J, Jauregui I, Davila I, Ferrer M, Bartra J, et al. Stress and allergy. J Investig Allergol Clin Immunol 2009;19(Suppl.1):40–7.
Dronjak S, Jezova D, Kvetnansky R. Different effects of novel stressors on sympathoadrenal system activation in rats exposed to long term immobilization. Ann NY Acad Sci 2004;1018:113–23.
Agarwal A, Prabakaran SA. Mechanism, measurement, and prevention of oxidative stress in male reproductive physiology. Indian J Exp Biol 2005;43:963–74.
Enstrom JE. Vitamin C in prospective epidemiological studies. In: Packer L, Fuchs J, editors. Vitamin C in health and disease. New York: Marcel Dekker Inc; 1997.p. 381–98.
Gey KF. Vitamins E plus C and interacting conutrients required for optimal health. Biofactors 1998;7:113–74.
Lodhi GM, Butt IF, Hussain MM, Aslam M. Effects of ascorbic acid and selenium supplementation on basal testosterone cortisol ratio in male Sprague Dawley rats. Pak Armed Forces Med J 2009;59(1):14–9.
Dillard CJ, Kunert KJ, Tappel AL. Effects of vitamin E, ascorbic acid and mannitol on alloxan-induced lipid peroxidation in rats. Arch Biochem Biophys 1982;216:204–12.
Ergul Y, Erkan T, Uzun H, Genc H, Altug T, Erginoz E. Effect of vitamin C on oxidative liver injury due to isoniazid in rats. Pediatr Int 2010;52:69–74.
Sonmez M, Turk G, Yuce A. The effect of ascorbic acid supplementation on sperm quality, lipid peroxidation and testosterone levels of male Wistar rats. Theriogenology 2005;63:2063–72.
Kabay SC, Ozden H, Guven G, Ustuner MC, Degirmenci I, Olgun EG, et al. Protective effects of vitamin E on central nervous system in streptozotacin induced diabetic rats. Clin Invest Med 2009;32:E314–21.
Akturk O, Demirin H, Sutcu R, Yilmaz N, Koylu H, Altuntas I. The effects of diazinon on lipid peroxidation and antioxidant enzymes in rat heart and ameliorating role of vitamin E and vitamin C. Cell Biol Toxicol 2006;22:455–61.
Judge AR, Selsby JT, Dodd SL. Antioxidants attenuate oxidative damage in rat skeletal muscle during mild ischaemia. Exp Physiol 2007;93:479–85.
Huang HY, Appel LJ, Croft KD Miller ER 3rd, Mori TA, Puddey IB. Effects of vitamin C and vitamin E on in vivo lipid peroxidation: results of a randomized controlled trial. Am J Clin. Nutr 2002;76:549–55.
Halpner AD, Handelman GJ, Harris JM, Belmont CA, Blumberg JB. Protection by Vitamin C of Loss of Vitamin E in Cultured Rat Hepatocytes. Arch Biochem Biophys 1998;359:305–9.
Lodhi GM, Hussain MM, Aslam M. Effect of antioxidant supplementation on lipid peroxidation and antioxidant enzyme activity on exposure to acute restraint stress in Sprague Dawley rats. Pak Armed Forces Med J 2012;62:105–10.
Nadeem A, Masood A, Masood N, Gilani RA, Shah ZA. Immobilization stress causes extracellular oxidant- antioxidant imbalance in rats: restoration by L-NAME and vitamin E. Eur Neuropsychopharmacology 2006;16:260–7.
Sim AS, Salonikas C, Naidoo D, Wilcken DE. Improved method for plasma malondialdehyde measurement by high performance liquid chromatography using methyl malondialdehyde as an internal standard. J Chromatogr B Analyt Technol Biomed Life Sci 2003;785:337–44