Cover
Vol. 5 No. 1 (2015)

Published: June 30, 2015

Pages: 14-19

Research Article

Effect of L-carnitine and Co Enzyme Q10 Addition to SMART Pro-Medium on Human Sperm Parameters During In Vitro Sperm Activation

Majida H. M. Al Shammari 1 Jabbar H. Yenzeel Muhammad-Baqir M-R. Fakhrildin 2
1Department of Biology-College of Science-University of Baghdad-Baghdad,IRAQ
2Department of Clinical Reproductive Physiology-High Institute of Infertility Diagnosis and Assisted Reproductive Technologies-Al-Nahrain University-Baghdad, IRAQ
History
  • Received: March 1, 2015
  • Available Online: June 1, 2015

Abstract

This study was designed to assess the effect of different concentrations of L-carnitine and Co enzyme Q10 (COQ10) supplied to SMART Pro-medium on human sperm motility and sperm grade activity ( progressive, non-progressive and immotile) during in vitro sperm activation. Eighty seven samples of semen of infertile and fertile men were randomly collected from the subjects at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies / Al-Nahrain University. The mean age of the men was (32.034 ± 0.57) years with mean duration of infertility (4.644 ± 0.25) years. Seminal fluid analyses were done involving macroscopic and microscopic examinations according to WHO criteria. Swim-up technique was dependent for in vitro sperm activation , the washed samples were divided before using centrifugation swim-up technique into 3 groups: control group (G1) without L-carnitine and CoQ10. While, in G2, G3 two concentrations of L-carnitine and CoQ10 (20 µg , and 40 µg ) were used. Moreover, all groups within post-activation enriched with SMART-Pro media. The sperm motility and sperm grade activity were evaluated after the addition of L-carnitine and COQ10. The results showed a significant (P<0.05) differences were showed in sperm motility and sperm grade activity of post-activation when compared with the pre-activation. In G2 and G3, with dose of ( 20 µg and 40 µg) of L-carnitine and CoQ10, the results showed a significant (P<0.05) increase in the percentages of sperm motility, progressive sperm motility in spite of the increase in the percentages of sperm motility, progressive sperm motility in G3 compared to G2, there was non significant (P>0.05) differences between G2 and G3. The results for non-progressive sperm motility% and total progressive sperm motility millions/ mL. for 40 µg ( G3) groups were significantly (P< 0.05) decreased when compared to control group (G1) with non-significant (P>0.05) differences between 20µg (G2) group, with significant (P< 0.05) decreased in immotile sperm percentage in post-activation when compared to the pre-activation. From the results of the study, it can be concluded that the addition of 40 µg of L-carnitine and CoQ10 to washed sperms can improve sperm motility and sperm grade activity in vitro. Data were analyzed statistically using complete randomized design (CRD) (one way ANOVA).

Keywords

References

  1. World Health Organization (WHO). (2010). WHO laboratory manual for the examination of human semen and semen-cervical mucus interaction. 5th edition. Cambridge: Cambridge University Press. pp:162-224.
  2. Tan, Y. ; and MJ Bennett (2007). The Australian & New Zealand journal of obstetrics and gynaecology, 47 (5): 406–9.
  3. Agarwal, H.S. ; K.S. Kulkarni. (2003). Efficacy and safety of semen in patient with oligospermia: an open clinical study. Indian J. Clin. Practice. 14 (2): 29-31.
  4. Francavilla, F.; F. Sciarretta; S. Sorgentone; S. Necozione; R. Santucci; A. Barbonetti; and S. Francavilla. (2009). Intra uterine insemination with or without mild ovarian stimulation in couples with male sub fertility due to oligo/asthenoand/ or teratozoospermia or anti sperm antibodies: a prospective cross-over trial. Fertil Steril, 92(3): 1009 -1011.
  5. Lewis, S.E.M. (2007). Is sperm evaluation useful in predicting human fertility? Reproduction , 134:1–11.
  6. Karpuz, V.; A .Gokturk; and M. Koyuturk. (2007). The Effect of Sperm Morphology and Motility on the Outcome of Intra cytoplasmic Sperm Injection. Marmara Med. J., 20 (2) ; 92-97.
  7. Isidori A.M.; C. Pozza; D. Gianfrilli; and A. Isidori. (2006). Medical treatment to improve sperm quality. Reprod Biomed Online, ; 12: 704 - 714.
  8. Lambardo, F.; L. Gandini; A. Lenzi; and F. Dondero. (2004). Anti sperm immunity in assisted reproduction. J Reprod Immunol. 62: 101-109.
  9. Pacey, A.A. (2006). Is quality assurance in semen analysis still really necessary? A view from the andrology laboratory. Hum Reprod; 21:11059.
  10. Matalliotakis, I.; Y. Koumantaki; and A. Evageliou. (2000). L-carnitine levels in the seminal plasma of fertile and infertile men: correlation with sperm quality. Int J Fertil Women. 45: 236–240.
  11. Lewin, A.; and H. Lavon. (1997). The effect of coenzyme Q10 on sperm motility and function. Mol Aspects Med, suppl. 18: S213-9.
  12. Bentinger, M. K. Brismar; and G. Dallner. (2007).The antioxidant role of coenzyme Q. Mitochondrion.7 Suppl:S41–S50.
  13. NAFA and ESHRE. (2002). Nordic Association for Andrology and European Society of Human Reproduction and Embryology- Special Interest Group on Andrology. Manual on Basic Semen Analysis. 1- 24.
  14. World Health Organization (WHO). (1999).WHO laboratory manual for the examination of human semen and semen-cervical mucus interaction. 4th edition. Cambridge: Cambridge University Press.
  15. Huntington, (2001). Study Group. A randomized, placebo controlled trial of coenzyme Q10 and re macemide in Huntington’s Disease. Neurology. 57:397-404.
  16. Miodrag, S.; W. Kirsten; Z. Valeri; S. Petra; B. Katja; and W. Eckhard. (1999). Coenzyme Q10 in Submicron- Sized Dispersion Improves Development, Hatching, Cell Proliferation, and Adenosine Tri phosphate Content of In Vitro- Produced Bovine Embryos1 biology of reproduction. 61, 541–547.
  17. Grane, F.L. (2001). Biochemical functions of coenzyme Q10J Am Coll Nutr. 20 (6):591-598.
  18. Mizuno, K.; M. Tanaka; S. Nozaki; H. Mizuma; S. Ataka; and T. Tahara, et al. (2008). Anti fatigue effects of coenzyme Q10 during physical fatigue. Nutrition . 24: 293-9.
  19. Reynolds, J.A.; and S.C. Hand. (2004). Differences in isolated mitochondria are insufficient to account for respiratory depression during diapause in art emiafranciscana embryos. Physiol. Bio chem. Zool. 77, 366-377.
  20. Levavasseur, F.; H. Miyadera; J. Sirois; M.L. Tremblay; K. Kita; and E. Shoubridge, et al. (2001). Ubiquinone is necessary for mouse embryonic development but is not essential for mitochondrial respiration. The Journal of Biological Chemistry, 276, 46160– 46164.
  21. Dallner, G. ; and PJ .Sindelar. (2000). Regulation of ubiquinone metabolism. Free Radic Biol Med. 29:285–294.
  22. Henkel, R. and Schill, W. (2003). Sperm preparation for ART. Repord Biol. And Endocrinol. 1:108-120.
  23. Alvarez JG, Lasso JL, Blasco L, Nunez RC, Heyner S, Caballero PP, et al. (1993). Centrifugation of human spermatozoa induces sub lethal damage; separation of human spermatozoa from seminal plasma by a dextran swim-up procedure without centrifugation extends their motile lifetime. Hum Reprod. 8:1087–92.
  24. Shaaban, M. H. (2007). An in vitro human sperm activation study: using Hams F-12 medium and human serum albumin for infertile patients. MS.C. Thesis. Institute of embryo research and infertile treatment Al-Nahrain university. Pp 102.
  25. Stagimiller, R.B.; and R.M.. Moor. (1984). Effect of follicle cells on the maturation and development competence of ovine oocyte matured outside the follicle. Gamete Res. 9:221-229.
  26. Trounson, A.; and A. Caro. (1984). Research in human in vitro fertilization and embryo transfer. Br., Med. J. 285: 244.
  27. Makler, A.; M. Fisher; O. Murillo; N. Laufer; A. Dechereny ; and F. Naftilin.(1984). Factor affecting sperm motility. IX. Survival of spermatozoa in various biological media and under different gaseous compositions. Fertil. Steril. 41: 428-432.
  28. Fakhrildin, M.B.; and N. K. Flayyih. (2011). A new simple medium for in vitro sperm activation of asthenozoospermic patients using direct swim-up technique. Kufa Med. Journal. 14(1): 67-75.
  29. King, RS.; and GJ. Killian. ( 1994). Purification of bovine estrus associated protein and localization of binding on sperm. Biol Reprod . 51: 34–42.
  30. Mortimer, D. (2000). Sperm preparation methods. Andrology lab cancer. J. Androl. 21:334-340.
  31. Balercia G; F. Regoli; T. Armeni; A .Koverech; and F .Mantero. (2005). Boscaro Placebo-controlled double- blind randomized trial on the use of L- carnitine, L-acetylcarnitine, or combined L-carnitine and L-M. acetylcarnitine in men with idiopathic asthenozoospermia. Fertil Steril 84:662–71.
  32. Al-Dujaily, S.S.; N. Al-Shahiri ; and A. Abbas. (2012). Effect of L- carnitine on in vitro sperm activation of infertile men. I J Embryo inter Res. 2(3): 22-25.
  33. Xuan, W.; A.M. Lambonwah ; C. Librach ; K. Jarvi and I. Tein. (2003) . Characterization of organic cation/carnitine transporter family in human sperm. Bioch. Biophys. Res. Com., 306: 121-128.
  34. Agarwal, A.; and T.M. Said.(2004). Carnitine and male infertiity. Reprod. Biomed. 8:376-384. Iraqi Journal of Embryo and Infertility Researches Vol.(5) Special Issue (2015) 19
  35. Gülçin, I. (2006). Antioxidant and antiradical activities of L-carnitine. Life Sci. 78:803-811.
  36. Solarska , K. ; A. Lewińska ; A. Karowicz-Bilińska ; G. Bartosz. ( 2010). The antioxidant properties of carnitine in vitro. Cell Mol Biol Lett 15: 90-97.
  37. López, L.; C. Quinzii; E. Area; A. Naini; S. Rahman; M. Schuelke; L. Salviati; S. Dimauro; and M. Hirano. (2010). Treatment of CoQ10 deficient, fibroblasts with Ubiquinone, CoQ analogs, and vitamin C:time-and compound-dependent effects. PLoS One, 5(7): 897-903.
  38. Balercia, G.; E. Buldreghini; A. Vignini; L. Tiano; F. Paggi; S. Amoroso; R. Ricciardo-Lamonica; M. Boscaro; A. Lenzi; and G. Littarru. (2009). Coenzyme Q10 treatment in infertile men with idiopathic asthenozoospermia: a placebo- controlled, double-blind randomized trial. Fertil Sterility 91:1785–1792.
  39. Mancini, A.; and G. Balercia .( 2011). Coenzyme Q(10) in male infertility: physiopathology and therapy. Biofactors. 37:374–380.