Abstract
The present study was designated to assess oxidative damage and its effect on germ cell apoptosis in testes of streptozotocin (STZ)-induced diabetic rats. The role of antioxidant supplementation with a mixture of vitamins E and C and alpha lipoic acid for protection against such damage was also evaluated. Forty-five adult male rats were randomly divided into three groups: group I, control, non-diabetic rats; group II, STZ-induced, untreated diabetic rats; group III, STZ-induced diabetic rats supplemented with a mixture of vitamins E and C and alpha lipoic acid. Glycated hemoglobin (HbA1C), glucose, and insulin levels were estimated in blood samples. Malondialdehyde (MDA), the activities of the enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx), and caspase-3 in addition to testosterone (T) level were all determined in testicular tissues. Histopathological studies using H&E stain, as well as, immunohistochemical detection of apoptosis using (TUNEL) method were also performed. Blood glucose and HbA1c were significantly increased while insulin was significantly decreased in STZ-induced diabetic rats as compared with controls. In rat testicular tissues, MDA, and caspase-3 activity were significantly elevated while SOD and GPx enzymatic activities as well as T level were significantly decreased in diabetic rats as compared with control group. Antioxidant supplementation to diabetic rats restored the testicular enzymatic activities of SOD and GPx to almost control levels, in addition, MDA and caspase-3 activity decrease while T increase significantly as compared with untreated diabetic group. Prominent reduction of germ cell apoptosis was found in diabetic rats supplemented with antioxidants. An important role of testicular oxidative damage and germ cell apoptosis in diabetes-induced infertility could be suggested, treatment with antioxidants has a protective effect by restoring SOD and GPx antioxidant enzymatic activity.
Similar content being viewed by others
References
Amos AF, Mccarty DJ, Zimmet P (1997) The rising global burden of diabetes and its complications: estimates and projections to the year 2010. Diabet Med 14:S1–S85
Bannister JU, Bannister WH, Rotilio G (1987) Aspects of the structure, function, and applications of superoxide dismutase. CRC Crit Rev Biocheme 22:111–180
Bonnefont R (2004) The role of antioxidant micronutrients in the prevention of diabetic complications. Treat Endocrinol 3:41–52
Bonnefont R, Bastard JP, Jaudon MC, Delattre J (2000) Consequences of diabetic status on the oxidant/antioxidant balance. Diab Metab 26:163–176
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Brigelius FR (1999) Tissue specific functions of individual glutathione peroxidases. Free Radic Biol Med 27:951–965
Brownlee M (1995) The pathological implications of proteins glycation. Clin Invest Med 18:275–281
Budin SB, Othman F, Louis SR, Abu Bakar M, Radzi M, Osman K (2009) Effect of alpha lipoic acid on oxidative stress and vascular wall of diabetic rats. RJME 50:23–30
Bursell SE, King GL (1999) Can protein kinas C inhibition and vitamin E prevent the development of diabetic vascular complications? Diab Res Clin Pract 5:169–182
Cay M, Nazıro M, Sim sek H, Aydilek N, Aksakal M, Demirci M (2001) Effects of intraperitoneally administered vitamin C on antioxidative defence mechanism in streptozotocin-induced diabetic rats. Res Exp Med 200:205–213
Coleman MD, Fernandes S, Khanderia L (2003) A preliminary evaluation of a novel method to monitor a triple antioxidant combination (vitamins E, C and alpha-lipoic acid) in diabetic volunteers using in vitro methaemoglobin formation. Environ Toxicol Pharmacol 14:69–75
D’ Aniello A, Cosmo AD, Cristo CD, Assisi L, Virgilo B, Fiore MD (1996) Occurrence of sex steroid hormones and their binding proteins in octopus vulgaris lam. Biochem Biophys Res Commun 277:782–788
Dickinson PJ, Carrington AL, Frost GS, Boulton AJM (2002) Neurovascular disease, antioxidants and glycation in diabetes. Diab Metab Res Rev 18:260–272
Erkkilä K, Henriksén K, Hirvonen V, Rannikko S, Salo J, Parvinen M, Dunkel L (1997) Testosterone regulates apoptosis in adult human seminiferous tubules in vitro. J Clin Endocrinol Metab 82:2314–2321
Fadeel B, Orrenius S (2005) Apoptosis: a basic biological phenomenon with wide-ranging implications in human disease. J Intern Med 58:479–517
Flohe L, Gunzler WA (1984) Assay of glutathione peroxidase. Meth Enzymol 105:114–121
Flora SJ (2007) Role of free radicals and antioxidants in health and disease. Cell Mol Biol 53:1–2
Fujii J, Luchi Y, Matsuki S, Lshii T (2003) Cooperative function of antioxidant and redox systems against oxidative stress in male reproductive tissues. Asian J Androl 5:231–242
Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493–501
Gillery P (2006) Oxidative stress and protein glycation in diabetes mellitus. Ann Biol Clin Paris 64:309–314
Glenn DR, McClure N, Lewis SE (2003) The hidden impact of diabetes on male sexual dysfunction and fertility. Hum Fertil Camb 6:174–179
Gnodos B, Rivikind Y, Jovaniivic L (1998) Effect of increasing glucose concentration on sertoli cell viability in the non obese diabetic mouse (NOD mouse). Ann Clin Lab Sci 28:236–241
Ihara Y, Yamada Y, Toyokuni S, Miyawaki K, BanN AT, Kuroe A, Iwakura T, Kubota A, Hiai H, Seino Y (2000) Antioxidant alpha-tocopherol ameliorates glycemic control of GK rats, a model of type 2 diabetes. FEBS Lett 473:24–26
Jones DP (2006) Redefining oxidative stress. Antioxid Redox Signal 8:1865–1879
Kagan VE, Serbinova EA, Forte T, Scita G, Paker L (1992) Recycling of vitamin E in human low density lipoproteins. J Lipid Res 33:385–397
Khokha AM, Kashko MF, Voronov PP (1993) Effect of ethanol and lipid peroxidation on testosterone biosynthesis by interstitial cells of testes. Ukr Biochem Zh 65:111–115
Konrad D, Somwar R, Sweeney G, Yaworsky K, Hayashi M, Ramlal T, Klip A (2001) The antihyperglycemic drug α-lipoic acid stimulates glucose uptake via both GLUT4 translocation and GLUT4 activation: potential role of p38 mitogen-activated protein kinase in GLUT4 activation. Diabetes 50:1464–1471
Lee JS, Ahn SS, Jung KC, Kim Y-W, Lee SK (2004) Effects of 60 Hz electromagnetic field exposure on testicular germ cell apoptosis in mice. Asian J Androl 6:29–34
Marklund S, Marklund G (1974) Involvement of superoxide anion radical in autoxidation of pyrogallol and convenient assay for superoxide dismutase. Eur J Biochem 47:469–474
Mostafa MH, Sharma RK, Thornton J, Mascha E, Abdel-Hafez MA, Thomas AJ (2004) Relationship between ROS production, apoptosis and DNA denaturation in spermatozoa from patients examined for infertility. Hum Reprod 19:129–138
Nagamatsu M, Nickander KK, Schmelzer JD, Raya A, Wittrock DA, Tritschler H, Low PA (1995) Lipoic acid improves nerve blood flow, reduces oxidative stress, and improves distal nerve conduction in experimental diabetic neuropathy. Diab Care 18:1160–1167
Ohkawa H, Ohishi N, Yagi K (1979) Assay of lipid peroxides in animal tissues by thiobarbituric acid reaction. Analyt Biochem 95:351–358
Opara EC (2002) Oxidative stress, micronutrients, diabetes mellitus and its complications. J R Soc Health 122:28–34
Packer L, Witt EH, Tritschler HJ (1995) Alpha-lipoic acid as a biological antioxidant. Free Radic Biol Med 19:227–250
Peirce EJ, Breed WG (2001) A comparative study of sperm production in two species of Australian arid zone rodents (Pseudomys australis, Notomys alexis) with marked differences in testis size. Reproduction 121:239–247
Robertson RP (2004) Chronic oxidative stress as a central mechanism for glucose toxicity in pancreatic islet beta cells in diabetes. J Biol Chem 279:42351–42354
Rosiepen G, Weinbauer GF, Schlatt S, Behre HM, Nieschlag E (1994) Duration of the cycle of the seminiferous epithelium estimated by the 5-bromodeoxyuridine technique, in laboratory and feral rats. J Reprod Fertil 100:299–306
Shaha C (2007) Modulators of spermatogenic cell survival. Soc Reprod Fertil Suppl 63:173–186
Shalaby MA, El Zorba HY, Kamel GM (2004) Effect of α-tocopherol and simvastatin on male fertility in hypercholesterolemic rats. Pharmacol Res 50:137–142
Shi Y (2002) Mechanisms of caspase activation and inhibition during apoptosis. Molec Cell 9:459–470
Shrilatha B, Muralidhara (2007) Occurrence of oxidative impairments, response of antioxidant defenses and associated biochemical perturbations in male reproductive milieu in streptozotocin-diabetic rat. Int J Androl 30:508–518
Soudamani S, Yuvaraj S, Malini T, Balasubramanian K (2005) Experimental diabetes has adverse effects on the differentiation of ventral prostate during sexual maturation of rats. Anat Rec Discov Mol Cell Evol Biol 287:1281–1289
Starin J (1991) Disturbances of micronutrients and antioxidant status in diabetes. Proc Nutr Soc 50:591–604
Sudha S, Valli G, Julie PM, Arunakaran J, Govindarajulu P, Balasubramanian K (2000) Influence of streptozotocin-induced diabetes and insulin treatment on the pituitary-testicular axis during sexual maturation in rats. Exp Clin Endocrinol Diab 108:14–20
Thulesen J, Orskov C, Holst JJ, Poulsen SS (1997) Short-term insulin treatment prevents the diabetogenic action of streptozotocin in rats. Endocrinology 138:62–68
Vessal M, Hemmati M, Vasei M (2003) Antidiabetic effects of quercetin in streptozotocin-induced diabetic rats. Comp Biochem Physiol C Toxicol Pharmacol 135C(3):357–364
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mohasseb, M., Ebied, S., Yehia, M.A.H. et al. Testicular oxidative damage and role of combined antioxidant supplementation in experimental diabetic rats. J Physiol Biochem 67, 185–194 (2011). https://doi.org/10.1007/s13105-010-0062-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13105-010-0062-2