Environmental pollutants such as di-(2-ethylhexyl) phthalate (DEHP) adversely affect reproductive system tissue differentiation and functions with exposure at intrauterine, neonatal or adult stages of life, thereby potentiating male infertility later in life. World health organization estimates a global infertility prevalent rate of 10-15%, and 20-30% among Nigerians, with male factor constituting about 40-50% of infertility cases. This study was designed to investigate the effect(s) of oral vitamins C and E on DEHP induced changes in some semen parameters and serum testosterone concentration in adult Wistar rats. Seventy (70) adult male Wistar rats weighing between 156-250 g were randomised into 7 experimental groups 1, 2, 3, 4, 5, 6 and 7 (group n=10). Animals in groups 1, 2 and 3 were treated with 0.02 mg, 20 mg, 200 mg oral DEHP/kg bw daily respectively, while those in groups 4, 5 and 6, in addition to the above DEHP treatments, were treated with 100 mg ascorbic acid and 67.5 mg α-tocopherol per kg bw daily respectively. Rats in group 7 served as Control and were treated with vehicle. All treatments lasted for 60 days. After, over night fasting, samples of semen and serum were obtained for analysis. Results obtained were expressed as mean ± standard deviation and analyzed for significant differences in means using one way ANOVA and Post Hoc test. Relative to the control reference values, groups exposed to oral DEHP had significant (p<0.05) reduction in sperm count, total sperm motility, active sperm motility, normal sperm morphology, serum testosterone concentration and serum super oxide dismutase levels to 31.70±18.68x106 cells/mL, 38.60±24.78%, 8.50±5.66%, 38.00±18.00%, 9.56±1.34 ng/mL and 0.017±0.0013 units respectively. Sluggish sperm motility and abnormal sperm morphology significantly (p˂0.01) increased to 39.70±13.05% and 68.50±18.42% respectively. In the groups that had DEHP co-treatments with oral ascorbic acid and α-tocopherol, all studied parameters tended to comparative indifference statistically, with the Controls values. This indicates a protective function against DEHP effects on the studied parameters. The study has shown therefore, that DEHP inflicts oxidative stress in the reproductive system which potentially suppresses serum testosterone concentration with attendant derangements in the qualitative and quantitative sperm cells in adult Wistar rats, and thereby enhancing male infertility. However, the antioxidants ascorbic acid and α-tocopherol protects the gonadal and sperm cells from the harmful effects of DEHP by ameliorating oxidative stress and improving male fertility. This implies that there is need to avoid prolonged exposure to DEHP while encouraging the daily intake of oral ascorbic acid and α-tocopherol.
Jensen, T. K., Sobotka, T., Hansen, M. A., Pedersen, A. T., Lutz, W., Skakkebaek, N. E.. (2008). Declining trends in conception rates in recent birth cohorts of native Danish women: a possible role of deteriorating male reproductive health. International Journal of Andrology 31(2): 81–92.
Agarwal, A., Mulgund, A., Hamada, A., & Chyatte, M. R. (2015). A unique view on male infertility around the globe. Reproductive Biology and Endocrinology: RB&E, 13, 37. http://doi.org/10.1186/s12958-015-0032-1
WHO. Infecundity, infertility, and childlessness in developing countries. DHS Comparative Reports No 9. Calverton, Maryland, USA. 2004 ORC Macro and the World Health Organization.
Sundby, J., Mboge, R., & Sonko, S. (1998). Infertility in the Gambia: frequency and health care seeking. Social Sciences and Medicine, 46, 91–99.
Geelhoed, D. W., Nayembil, D., Asare, K., S., van Leeuwen, J., H., & Roosmalen, J. (2002). Infertility in rural Ghana. International Journal of Gynaecology and Obstetrics, 79, 137–142.
Haile, A. (1990). Fertility conditions in Gondar, northwestern Ethiopia: an appraisal of current status. Studies in Family Planning, 21, 110–118.
Adeniji, R. A., Olayemi, O., Okunlola, M.A., & Aimakhu, C.O. (2003). Pattern of semen analysis of male partners of infertile couples at the University College Hospital, Ibadan. West African Journal of Medicine, 2003; 22: 243–245.
Uadia, P. O., & Emokpae, A. M. (2015). Male infertility in Nigeria: A neglected reproductive health issue requiring attention. Journal of Basic and Clinical Reproductive Science. 2015; 4(2): 45-55.
Panti. A. A., & Sununu, Y. T. (2014). The profile of infertility in a teaching Hospital in North West Nigeria. Sahel Medical Journal, 17: 7-11..
Sokol, Z. R., Kraft, P., Fowler, I. M., Mamet, R., Kim, E., & Berhane, K. T. (2006). Exposure to environmental Ozone alters semen quality. Environmental Health Perspectives, 114(3), 360–365.
Baralić, K., Buha Djordjevic, A., Živančević, K., Antonijević, E., Anđelković, M., Javorac, D., Ćurčić, M., Bulat, Z., Antonijević, B., & Đukić-Ćosić, D. (2020). Toxic Effects of the Mixture of Phthalates and Bisphenol A-Subacute Oral Toxicity Study in Wistar Rats. International journal of environmental research and public health, 17(3), 746. https://doi.org/10.3390/ijerph17030746
Ogli, S. A., Odeh, S. O., & Egesie, G. U. (2019). Gonadotoxic Potentials of Di- (2-Ethyl Hexyl) Phthalate in the Adult Male Wistar Rats. Journal of Research in Basic Clinical Sciemce, 2: 163–171.
Telisman, S., Cvitkovic, P., Jurasovic, J., Pizent, A., Gavella, M., & Rocic, B. (2001). Semen quality and reproductive endocrine function in relation to biomarkers of lead, cadmium, zinc, and copper in men. Environmental Health Perspectives, 108, 45–53.
Ma, T., Zhou, W., Chen, L., Wu, L., Christie, P., Zhang, H., et al. (2017) Toxicity effects of di-(2-ethylhexyl) phthalate to Eisenia fetida at enzyme, cellular and genetic levels. PLOS ONE 12(3): 1-12. e0173957. https://doi.org/10.1371/journal
Food and Drug Administration (FDA) (2002). Public health notification: PVC devices containing the plasticizer DEHP, US. FDA. Center for Devices and Radiological Health. Retrieved June 10th, 2006 from: https://www.fda.gov/cdrh/safety/dehp/html.
Quinnies, K. M., Harris, E. P., Snyder, R. W., Sumner, S. S., & Rissman, E. F. (2017). Direct and transgenerational effects of low doses of perinatal di-(2-ethylhexyl) phthalate (DEHP) on social behaviors in mice. PLOS ONE, 12(2), e0171977. https://doi.org/10.1371/journal.pone.0171977
Ge, R., Chen, G., Dong, Q., Akingbemi, B., Sottas, C. M., Santos, M., Sealfon, S. C., Bernard, D. J., & Hardy, M. P. (2007). Biphasic Effects of postnatal exposure to Diethylhexylphthalate on the timing of puberty in male Rats. Journal of Andrology, 28(4), 513-520.
Hansen, C., Luben, T. J., Sacks, J. D., Olshan, A., Jeffay, S., Strader, L., & Perreault, S. D. (2010). Effect of ambient air pollution on sperm quality. Environmental Health Perspectives, 118(2), 203-209.
Dai, X., Zhang, Q., Yu, Y, Sun, W., Wang, R., & Miao, D. (2018). Bmi1 Deficient Mice Exhibit Male Infertility. International Journal of Biological Sciences, 14(3), 358–368.
Petroianu, A., & Alberti, L. R. (2011). Effect of oral supplementation of vitamin C on intestinal anastomotic resistance. Revista do Colegio Brasiliero de Cirurgioes, 38(1), 54-58
Heinrich, U. R., Fischer, I., Brieger, J., Rümelin, A., Schmidtmann, I., Li, H., Mann, W. J., & Helling, K. (2008). Ascorbic acid reduces noise-induced nitric oxide production in the guinea pig ear. Laryngoscope, 118(5), 837-842.
Takhshid, M. A., Tavasuli, A. R., Heidary, Y., Keshavarz, M., & Kargar, H. (2012). Protective Effect of Vitamins E and C on Endosulfan-Induced Reproductive Toxicity in Male Rats. Iranian Journal of Medical Sciences, 37(3), 173–180.
Ogli, S. A., Ochalefu, D. O., & Ojabo, C. O. (2011). Effects of oral supplements regime of vitamins C and E on sperm count in Wistar rats. Journal of Medicine in the Tropics, 13, 50-53.
Tang, X., Wu, S., Shen, L., Wei, Y., Cao, X., Wang, Y., Long, C., Zhou, Y., Li, D., Huang, F., Liu, B., & Wei, G. (2018). Di-(2-ethylhexyl) phthalate (DEHP)-induced testicular toxicity through Nrf2-mediated Notch1 signaling pathway in Sprague-Dawley rats. Environmental Toxicology, 33(7), 720-728.
Adewoyin, M., Ibrahim, M., Roszaman, R., Md Isa, M. L., Mat Alewi, N. A., Abdul Rafa, A. A., & Anuar, M. N. N. (2017). Male Infertility: The Effect of Natural Antioxidants and Phytocompounds on Seminal Oxidative Stress. Diseases, 5(1), 9. http://doi.org/10.3390/diseases5010009.
Rajesh, P., Sathish, S., Srinivasan, C., Selvaraj, J., & Balasubramanian, K. (2013). Phthalate is associated with insulin resistance in adipose tissue of male rat: role of antioxidant vitamins. Journal of Cellular Biochemistry, 114(3), 558-569.
Animal Environment, Housing, and Managements (1996). In: D. Clark (Rev). Guide for the Care and Use of Laboratory Animals (pp. 21-25). Washington, DC: The National Academies Press.
Takano, H., Yanagisawa, R., Inoue, K. I., Ichinose, T., Sadakano, K., & Yoshikawa, T. (2006). Di-(2-ehylhexyl) Phthalate enhances atopic dermatitis-like skin lesions in Mice. Environmental Health Perspectives, 114, 1266-1269.
Badr, G., Bashandy, S., Ebaid, H., Mohany, M., & Sayed, D. (2012). Vitamin C supplementation reconstitutes polyclonal T cells in streptozotocin-induced diabetic rats. European Journal of Nutrition, 52(5), 623-633.
Petroianu, A., & Alberti, L. R. (2011). Effect of oral supplementation of vitamin C on intestinal anastomotic resistance. Revista do Colegio Brasiliero de Cirurgioes, 38(1), 54-58.
Krishnamoorthy, G., Venkataraman, P., Arunkumar, A., Vignesh, R. C., Aruidhas, M. M., & Arunakaran, J. (2007). Ameliorative effect of vitamins (α-tocopherol and ascorbic acid) on PCB (Aroclor 1254) induced oxidative stress in rat epididymal sperm Reproductive Toxicology, 23(2), 239-245.
Biswas, A., Mohan, J., Sastry, K. V., & Tyagi, J. S. (2007). Effectiveness of dietary vitamin E on the cloacal gland, foam and semen characteristics of male Japanese quail. Theriogenology, 67(2), 259-263.
University of Delaware, Office of Laboratory Animal Medicine. Cardiac puncture blood collection (Terminal Procedure) SOP #PRO-002. Rev.May 2010. Rerieved on May 20th, 2011 from: http://www.udel.edu/research/pdf/Cardiac-Puncture-Blood-Collection-Terminal.pdf.
Cheesbrough M. (2000). Examination of semen. District Laboratory Practice in Tropical Countries (2nd Low price. ed., part II). 2000; pp. 130-132. Cambridge UK; University press.
Sikka, S. C., Hellstrom, W. J. (2016). Current updates on laboratory techniques for the diagnosis of male reproductive failure. Asian Journal of Andrology, 18(3): 392–401.
Van Haaster, L. H., & de Rooij, D. G. (1993). Spermatogenesis is accelerated in the immature djungarian and chinese hamster and rat. Biology of Reproduction, 49, 1229-1235.
Marklund, S., & Marklund, G. (1974). Involvement of the Superoxide Anion Radical in the Autoxidation of Pyrogallol and a Convenient Assay for Superoxide Dismutase European Journal of Biochemistry, 47, 469-474.
Gavali, Y., Deore, D., Surwase, S. P., & Zingade, U. (2013). Study of the serum superoxide dismutase levels in smoking and non-smoking patients with COPD. International Journal of Recent Trend in Science and Technology, 5(3), 121-126.
Wu, M., Xu, L., Teng, C., Xiao, X., Hu, W., Chen, J., & Tu, W. (2019). Involvement of oxidative stress in di-2-ethylhexyl phthalate (DEHP)-induced apoptosis of mouse NE-4C neural stem cells. Neurotoxicology, 70, 41–47. https://doi.org/10.1016/j.neuro.2018.10.013
Erkekoglu, P., Giray, B., Rachidi, W., Hininger-Favier, I., Roussel, A. M., Favier, A., & Hincal. F. (2011). Effects of di(2-ethylhexyl)phthalate on testicular oxidant/antioxidant status in selenium-deficient and selenium-supplemented rats. Environmental Toxicology, 10, 1002.
Culty, M., & Thuillier, R., Li, M., Wang, Y., Martinez-Arguelles, D. B., Benjamin, C. G., et al. (2008). In Utero exposure to Di-(2-ethylhexyl) phthalate exerts both short-term and long-lasting suppressive effects on testosterone production in the Rat. Biology of Reproduction, 78(6):1018–1028, https://doi.org/10.1095/biolreprod.107.065649.
Oishi, S., & Hiraga, K. (1980). Testicular atrophy induced by phthalic acid monoesters: effects of zinc and testosterone concentrations. Toxicology, 15, 197-202
Shi, Y. Q., Fu, G. Q., Zhao, J., Cheng, S. Z., Li, Y., Yi, L. N., Li, Z., Zhang, L., Zhang, Z. B., Dai, J., & Zhang, D. Y. (2019). Di(2-ethylhexyl) phthalate induces reproductive toxicity via JAZF1/TR4 pathway and oxidative stress in pubertal male rats. Toxicology and industrial health, 35(3): 228–238. https://doi.org/10.1177/0748233718824911
Botelho, G. G., Bufalo, A. C., Boareto, A. C., Muller, J. C., Morais, R. N., Martino-Andrade, A. J., et al. (2009). Vitamin C and resveratrol supplementation to rat dams treated with di(2-ethylhexyl)phthalate: impact on reproductive and oxidative stress end points in male offspring. Archives of Environmental Contamination and Toxicology.9; 57(4):785-793
Gill-Sharma, M. K. (2018). Testosterone Retention Mechanism in Sertoli Cells: A Biochemical Perspective. The Open Biochemistry Journal, 12: 103–112.
Ma T, Zhou W, Chen L, Wu L, Christie P, Zhang H, et al. (2017) Toxicity effects of di-(2-ethylhexyl) phthalate to Eisenia fetida at enzyme, cellular and genetic levels. PLOS ONE; 12(3): 1-12. e0173957.
Leung, P. S., Sernia C. (2003). The renin-angiotensin system and male reproduction: new functions for old hormones. Journal of Molecular Endocrinology, 30; 263–270.
Elliott, B. M., & Elcombe, C. R. (1987). Lack of DNA damage or lipid peroxidation measured in vivo in the rat liver following treatment with peroxisomal proliferators. Carcinogenesis, 8, 1213–1218.
Orbea, A., Ortiz-Zarragoitia, M., & Cajaraville, M. P. (2008). Interactive effects of benzo(a)pyrene and cadmium and effects of di(2-ethylhexyl) phthalate on antioxidant and peroxisomal enzymes and peroxisomal volume density in the digestive gland of mussel Mytilus galloprovincialis Lmk. Journal of Biomarkers, 7(1), 33-48.
Gong, P., Xu, H., Zhang, H., & Wang, Z. (2012). PPAR expression and its association with SOD and NF-κB in rats with obstructive jaundice. Biomedical Research-India, 23(4), 551-560.
Ernst, J., Jann, J. C., Biemann, R., Koch, H. M., & Fischer, B. (2014). Effects of the environmental contaminants DEHP and TCDD on estradiol synthesis and aryl hydrocarbon receptor and peroxisome proliferator-activated receptor signalling in the human granulosa cell line KGN. Molecular Human Reproduction, 9, 919-928.
Levine, M., Rumsey, S. C., Wang, Y., Park , J. B., & Daruwala, R. (2000). Vitamin C. In M. H. (Ed.), Biochemical and physiological aspects of human nutrition pp 541–567. Philadelphia: W B Saunders.
Juan, M. C., Alicia, P., Oriana, B., Bing, Z., Roberto, S., & David, W. G. (2004). Vitamin C is a kinase inhibitor. Molecular and Cellular Biology, 24(15), 6645-6652.
Traber, M. G., & Atkinson, J. (2007). "Vitamin E, antioxidant and nothing more". Free Radical Biology and Medicine, 43(1), 4–15.
Yarube, S. U., Ayo, J. O., & Fatihu, M. Y. (2014). Harmful effects of ascorbic acid and α-tocopherol on male reproductive organs of rats chronically exposed to sodium nitrate. Journal of Medicine in the Tropics, 16(1), 5-8.
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