DNA fragmentation and oxidative stress compromise sperm motility and survival in late pregnancy exposure to omega-9 fatty acid in rats

Document Type : Original Article


Department of Physiology, College of Medicine of the University of Lagos, Lagos, Nigeria


Objective(s):The aim of this study was to evaluate the oxidative status and DNA integrity in testes of wistar rat offspring exposed to omega-9 monounsaturated (MUFA) at different times of late organogenesis.
Materials and Methods:Sixty female rats were divided into six groups of 10 animals. The first group served as control and received the drug vehicle, olive oil (1 ml/kg/day). The second, third, fourth, fifth and sixth group received 1000 mg/kg of oleic acid on gestation day 15 (D15), 16 (D16), 17 (D17), 18 (D18) and 19 (D19), respectively. Male pups were allowed to attain puberty and thereafter, blood was taken for hormonal analyses. Sperm count and motility were assessed. Testes homogenate was used for the determination of biochemical variables. Testes DNA was also determined.
Results:The results showed that sperm count and motility were significantly decreased in the treated groups as compared to the control. There was a marked increase in the malondialdehyde level in rat testes from all of the treated groups as compared to the control (P<0.05). DNA from the testes of rats of D19 had the highest level of fragmentation as compared to the control.
Conclusion: Omega-9 MUFA exposure in utero imposes negative effects on sperm variables and increases the level of sperm DNA fragmentation and oxidative stress.


1. Oyelowo OT, Raji Y, Bolarinwa AF. In –utero exposure to ripe Carica papaya seed resulted in postnatal outcomes from only first and second trimesters in rats. Int J Adv Res 2014; 2:254-266.
2. Win DT. Oleic acid –the anti-breast cancer component in olive oil. Au J T 2005; 9:75-78.
3. Arechaga G, Prieto I, Segarra AB, Alba F, Ruiz-Larrea MB, Ruiz-Sanz JI, et al. Dietary fatty acid composition affects aminopeptidase activities in the testes of mice. Int J Androl 2002; 25:113-118.
4. Safarinejad MR, Safarinejad S. The roles of omega-3 and omega-6 fatty acids in idiopathic male infertility. Asian J Androl 2012; 14:514–515.
5. Estienne MJ, Harper AF, Crawford RJ. Dietary supplementation with a source of omega-3 fatty acids increases sperm number and the duration of ejaculation in boars. Theriogenology 2008; 70:70–76.
6. Rooke JA, Shao CC, Speake BK. Effects of feeding tuna oil on the lipid composition of pig spermatozoa and In vitro characteristics of semen. Reproduction 2001; 121:315–322.
7. Conquer JA, Martin JB, Tummon I, Watson L, Tekpetey F. Effect of DHA supplementation on DHA status and sperm motility in asthenozoospermic males. Lipids 2000; 35:149–154.
8. Castellano CA, Audet I, Bailey JL, Chouinard PY, Laforest JP, Matte JJ. Effect of dietary n-3 fatty acids (fish oils) on boar reproduction and semen quality. J Anim Sci 2010; 88:2346–2355.
9. Weaver KL, Ivester P, Seeds M, Case LD, Arm JP. Effect of dietary fatty acids on inflammatory geneexpression in healthy humans. J Biol Chem 2009; 284:15400–15407.
10. Aksoy Y, Aksoy H, Altinkaynak K, Aydin HR, Ozkan A. Sperm fatty acid composition in subfertile men. Prostaglandins Leukot Essent Fatty Acids 2006; 75:75–79.
11. Oyelowo OT, Bolarinwa AF. Early-Life exposure to omega-9 fatty acid results in gonadal-regression in male rat (in press).
12. Comphaire FH, Mahmoud AM, Depuydt CE, Zalata AA, Christophe AB. Mechanisms and effects of male genital tract infection on sperm quality and fertilizing potential: The andrologist’s viewpoint. Hum Reprod Update 1999; 5:393-398.
13. Fraczek M, Kurpisz M. The redox system in human semen and peroxidative damage of spermatozoa. Postępy Hig Med Doś 2005; 59:523-534.
14. Sharma RK, Pasqualotto FF, Nelson DR, Thomas AJ, Agarwal A. The reactive oxygen species-total antioxidant capacity score is a new measure of oxidative stress to predict male infertility. Hum Reprod 1999; 14:2801-2807.
15. Takagi S, Itoh N, Kimura M, Sasao T, Tsukamoto T. Spermatogonial proliferation and apoptosis in hypospermatogenesis associated with nonobstructive-azoospermia. Fertil Steril 2001; 76:901-907.
16. Mallie JP, Boudzoumou P. Functional renal maturation in rats neonates after prenatal exposure to furosemide. Pediatr Nephrol 1996; 10:458-460.
17. Arafa HM, Aly HA, Abd-Ellah MF, El-Refaey HM. Hesperidin attenuates benzo[α] pyrene-induced testicular toxicity in rats via regulation of oxidant/antioxidant balance. Toxicol Ind Health 2009; 25:417.
18. Anderson RA Jr, Oswald C, Willis, BR, Zaneveld, LJ. Relationship between semen characteristics and fertility in electroejaculation mice. J Reprod Fertil 1983;68:1–7.
19. Cheever, KL, Weigel, WW, Rechards, DE, Lal, JB, Plotnick HB. Testicular effect of bis (2-methoxyethyl) ether in the adult male rat. Toxicol Ind Health 1989; 5:1099–1109.
20. Sun M, Zigman S. An improved spectrophotometric assay for superoxide dismutase based on epinephrine autoxidation. Anal Biochem 1978; 247:81–89.
21.  Aebi H. Catalase In vitro. Methods Enzymol 1984; 8:121–126.
22. van Dooran R, Liejdekker CM, Handerson PT. Synergistic effects of phorone on the hepatotoxicity of bromo-benzene and paracetamol in mice. Toxicology 1978; 11:225–233.
23. Uchiyama M, Mihara M. Determination of malonaldehyde precursor in tissues by thiobarbituris acid test. Anal Biochem 1978; 86:271–278.
24. Lowry OH, Rosenbrough NJ, Farr AI, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 1193:265–275.
25. Kim E, Goldnerg M. Direct method for cholesterol determination. Clin Chem 1969; 15:1171–1172.
26. Racevskis J. Molecular diagnosis of cancer: Methods and Protocols; second edition. Med Oncol 2005; 22:325-326.
27. Meistrich ML, Finch M, da Cunha MF, Hacker U, Au WW. Damaging effects of fourteen chemotherapeutic drugs on mouse testis cells. Cancer Res 1982; 42:122-131.
28. Inyang F, Ramesh A, Kopsombut P, Niaz MS, Hood DB, Nyanda AM. Disruption of testicular steroidogenesis and epididymal function by inhaled benzo(a)pyrene. Reprod Toxicol 2003; 17:527-537.
29. Gu Y, Davis DR, Lin YC. Developmental changes in lactate dehydrogenase-X activity in young jaundiced male rats. Arch Androl 1989; 22:131-136.
30. Koriem KM, Fathi GE, Salem HA, Akram NH, Gamil SA. Protective role of pectin against cadmium-induced testicular toxicity and oxidative stress in rats. Toxicol Mech Methods 2013; 23:263–272.
31. Abd-Allah AR, Aly HA, Moustafa AM, Abdel-Aziz AA, Hamada FM. Adverse testicular effects of somequinolone members in rats. Pharmacol Res 2000; 41:211–219.
32. Ramadan LA, Abd-Allah AR, Aly HA, Saad-el-Din AA. Testicular toxicity effects of magnetic field exposure and prophylactic role of coenzyme Q10 and L-carnitine in mice. Pharmacol Res 2002; 46:363–370.
33. Abdel-Wahab MH. Testicular toxicity of dibromoacetonitrile and possible protection by tertiary butylhydroquinone. Pharmacol Res 2003; 47:509–515.
34. Saradha B, Mathur PP. Effect of environmental contaminants on male reproduction. Environ Toxicol Pharmacol 2006; 21:34–41.
35. Veena BS, Sharmila U, Satish KA.  Evaluation of oxidative stress, antioxidants and prolactin in infertile women. Indian J Clin Biochem 2008; 23186-23190.
36. Mruk DD, Silvestrini B, Mo MY, Cheng CY. Antioxidant superoxide dismutase – a review: its function, regulation in the testis, and role in male fertility. Contra 2002; 67:305–311.  
37. Sies H. Strategies of antioxidants defense. Eur J Biochem 1993; 215:213–219.
38. Mommsen TP, Walsh PJ. Biochemical and environmental perspectives on nitrogen metabolism in fishes. Experientia1992; 48:583-593.
39. Yousef MI, Salama AF. Propolis protection from reproductive toxicity caused by aluminium chloride in male rats. Food Chem Toxicol 2009; 47:1168-1175.
40. Stocco D. Star protein and regulation of steroid hormone biosynthesis. Ann Rev Physiol 2001; 63:193–213.
41. Guyton AC, Hall JE. Reproductive and hormonal functions of the male (and the pineal gland). In Text book of medical physiology. 9th ed. Philadelphia (PA): Elseiver; 1998.p. 1003–1016.
42. Sharpe RM, Skakkeback NE. Are oestrogens involved in falling sperm counts and disorders of the male reproductive tract?  Lancet 1993; 341:1392-1395.
43. Kumar GP, Seerwani N, Laloraya M, Nivsarkar M, Verma S, Singh A. Superoxide dismutase as a regulatory switch in mammalian testicular steroidogenesis. Biochem Biophys Res Commun 1990; 173:302-308.
44. Narayana K. An aminoglycoside antibiotic gentamycin induces oxidative stress, reduces antioxidant reserve and impairs spermatogenesis in rats. J Toxicol Sci 2008; 33:85–96.
45. Taylor CT. Antioxidants and reactive oxygen species in human fertility.  Environ Toxicol Pharmacol 2001; 10:189-198.
46. Archibong AE, Ramesh A, Niaz, MS, Brooks CM, Roberson SI, Lunstra, DD.  Effects of benzo(a)pyr-ene on intra-testicular function in F-344 rats. Int J Environ Res Public Health 2008; 5:32–40.
47. Ochsendorf RF. Infections in the male genital tract and reactive oxygen species. Hum Reprod Update 1999; 5:399-320.
48. Lopes S, Sun JG, Jurisicova A, Meriano J, Casper RF. Sperm deoxyribonucleic acid fragmentation is increased in poor-quality semen samples and correlates with failed fertilization in intracytoplasmic sperm injection. Fertil Steril 1998; 69:528–532.
49. Smith M, Romijn JC, Wildhagen MF, Weber RF, Dohle GR. Sperm chromatin structure is associated with the quality of spermatogenesis in infertile patients Fertil Steril 2010; 94:1748–1752.
50. La Vignera S, Condorelli R, D’Agata R, Vicari E, Calogero AE. Semen alterations and flow-citometry evaluation in patients with male accessory gland infections. J Endocrinol Invest 2012; 35:219–223.
51. Erenpreiss J, Spano M, Erenpreisa J, Bungum M, Giwercman A. Sperm chromatin structure and male fertility: biological and clinical aspects. Asian J Androl 2006; 8:11–29.
52. Muratori M, Marchiani S, Maggi M, Forti G, Baldi E. Origin and biological significance of DNA fragmentation in human spermatozoa. Front Biosci 2006; 11:1491–1499.
53. Marzieh R, Ali R.T, Morteza A, Abolghasem AS, Marjan O. Saccharin consumption increases sperm DNA fragmentation and apoptosis in mice. Iran J Reprod Med 2014; 12:07-312.
54. Schulte RT, Ohl DA, Sigman M, Smith GD. Sperm DNA damage in male infertility: etiologies, assays, 
and outcomes.  J Assist Reprod Genet 2010; 27:3–12.
55. Tamburrino L, Marchiani S, Montoya M, Elia MF, Natali I, Cambi M, bet al. Mechanisms and clinical correlates of sperm DNA damage Asian J Androl  2012; 14:24–31.