Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Effects of L-carnitine and Pentoxifylline on the Activity of Lactate Dehydrogenase C4 isozyme and Motility of Testicular Spermatozoa in Mice

Authors
Elham Aliabadi 1
Fatemeh Karimi 1
Mozhgan Rasti 2
Masoumeh Akmali 2
Tahereh Esmaeilpour 1
1 Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
2 Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Abstract
Background: Extracted sperm from the testis have poor motility. Moreover, their motility changes during their journey through epidydimis. Meanwhile, they face high concentration of L-carnitin. In addition, lactate dehydrogenase C4 (LDH-C4) gene disorders has been shown to cause impaired sperm motility, leading to infertility in male mice. The aim of this study was to evaluate sperm motility and LDH-C4 ezyme activity upon L-carnitine (LC) and Pentoxifylline (PTX) administrations in mice. Methods: We extracted testicular sperm of 48 mice and divided them into three equal parts. One part was incubated with Ham's F10 medium (control), the other parts were treated with Ham's F10 containing LC and PTX with a final concentration of 1.76 mM, for 30 min at room temperature. Sperm motility was assessed according to the World Health Organization (WHO) criteria. Sperm LDH-C4 enzyme activity was measured by spectrophotometery method. Statistical analyses were performed using ANOVA and Fisher's LSD test, and a p-value less than 0.05 was considered as a statistically significant difference. Results: Sperm motility increased after 30 min of incubation in LC- and PTX-treated group (p < 0.001). LC and PTX administrations showed a significant increase in the LDHC4 enzyme activity of sperm compared to that of the controls after 30 min (P=0.04 and 0.01, respectively). Conclusion: The effects of LC and PTX on motility of sperm can be explained by an increase in LDH-C4 enzyme activity that may influence male fertility status. We suggest that LC as a non-toxic antioxidant is more suitable for use in assisted reproductive technique protocols than PTX.
Keywords
L
Carnitine
LDH
C4
Male infertility
pentoxifylline
Testicular sperm
  1. Isidori AM, Pozza C, Gianfrilli D, Isidori A. edical treatment to improve sperm quality. Reprod Biomed Online. 2006;12(6):704-14.
  2. Su LM, Palermo GD, Goldstein M, Veeck LL, Rosenwaks Z, Schlegel PN. Testicular sperm extraction with intracytoplasmic sperm injection for nonobstructive azoospermia: testicular histology can predict success of sperm retrieval. J Urol. 1999;161(1):112-6.
  3. Mann T, Lutwak-Mann C. Studies on the metabolism of semen. 4. Aerobic and anaerobic utilization of fructose by spermatozoa and seminal vesicles. Biochem J. 1948;43(2):266-70.
  4. Storey BT. Mammalian sperm metabolism: oxygen and sugar, friend and foe. Int J Dev Biol. 2008;52(5-6):427-37.
  5. Miki K, Qu W, Goulding EH, Willis WD, Bunch DO, Strader LF, et al. Glyceraldehyde 3-phosphate dehydrogenase-S, a sperm-specific glycolytic enzyme, is required for sperm motility and male fertility. Proc Natl Acad Sci U S A. 2004;101(47):16501-6.
  6. Li SS. Lactate dehydrogenase isoenzymes A (muscle), B (heart) and C (testis) of mammals and the genes coding for these enzymes. Biochem Soc Trans. 1989;17(2):304-7.
  7. Duan C, Goldberg E. Inhibition of lactate dehydrogenase C4 (LDH-C4) blocks capacitation of mouse sperm in vitro. Cytogenet Genome Res. 2003;103(3-4):352-9.
  8. Abd-Allah AR, Helal GK, Al-Yahya AA, Aleisa AM, Al-Rejaie SS, Al-Bakheet SA. Pro-inflammatory and oxidative stress pathways which compromise sperm motility and survival may be altered by L-carnitine. Oxid Med Cell Longev. 2009;2(2):73-81.
  9. Odet F, Duan C, Willis WD, Goulding EH, Kung A, Eddy EM, et al. Expression of the gene for mouse lactate dehydrogenase C (Ldhc) is required for male fertility. Biol Reprod. 2008;79(1):26-34.
  10. Sikka SC. Relative impact of oxidative stress on male reproductive function. Curr Med Chem. 2001;8(7):851-62.
  11. Chaudiere J. Some chemical and biochemical constraints of oxidative stress in living cells. In: Riee-Evans CA, Burdon RH, editors. Free radical damage and its control. Amsterdam: Elsevier; 1994. p. 25-66.
  12. Aitken RJ, Baker MA. Oxidative stress, sperm survival and fertility control. Mol Cell Endocrinol. 2006;250(1-2):66-9.
  13. Gil-Guzman E, Ollero M, Lopez MC, Sharma RK, Alvarez JG, Thomas AJ Jr, et al. Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation. Hum Reprod. 2001;16(9):1922-30.
  14. Lenzi A, Sgrò P, Salacone P, Paoli D, Gilio B, Lombardo F, et al. A placebo-controlled double-blind randomized trial of the use of combined l-carnitine and l-acetyl-carnitine treatment in men with asthenozoospermia. Fertil Steril. 2004;81(6): 1578-84.
  15. Aliabadi E, Soleimani-Mehranjani M, Borzoei Z, Talaei-Khozani T, Mirkhani H, Tabesh H. Effects of L-carnitine and L-acetyl-carnitine on testicular sperm motility and chromatin quality. Iran J Reprod Med. 2012;10(2):77-82.
  16. Kovacic B, Vlaisavljevic V, Reljic M. Clinical use of pentoxifylline for activation of immotile testicular sperm before ICSI in patients with azoospermia. J Androl. 2006;27(1):45-52.
  17. Tournaye H, Janssens R, Devroey P, van Steirteghem A. The influence of pentoxifylline on motility and viability of spermatozoa from normozoospermic semen samples. Int J Androl. 1994;17(1):1-8.
  18. Zare Z, Eimai H, Mohammadi M, Mofid M, dashtnavard H. The effect of orally administered l-carnitine on testis tissue, sperm parameters and daily sperm productionin adult mice. Cell J. 2010;11(4):382-9.
  19. Jenkins DL, Griffith OW. Antiketogenic and hypoglycemic effects of aminocarnitine and acylaminocarnitines. Proc Natl Acad Sci U S A. 1986;83(2):290-4.
  20. Beshay E, Croze F, Prud'homme GJ. The phosphodiesterase inhibitors pentoxifylline and rolipram suppress macrophage activation and nitric oxide production in vitro and in vivo. Clin Immunol. 2001;98(2):272-9.
  21. Gavella M, Lipovac V, Marotti T. Effect of pentoxifylline on superoxide anion production by human sperm. Int J Androl. 1991;14(5):320-7.
  22. Sato M, Ishikawa A. Room temperature storage of mouse epididymal spermatozoa: exploration of factors affecting sperm survival. Theriogenology. 2004;61(7-8):1455-69.
  23. Balaban B, Urman B, Isiklar A, Alatas C, Aksoy S, Mercan R, et al. Progression to the blastocyst stage of embryos derived from testicular round spermatids. Hum Reprod. 2000;15(6):1377-82.
  24. World Health Organization. WHO Laboratory manual for examination of human semen and sperm-cervical mucuinteraction. 4th ed. Cambridge: Cambridge University press; 2010.
  25. Moreira SG, Lipshultz LI. Management of male infertility. Sci World J. 2004;4:214-448.
  26. Wang YX, Yang SW, Qu CB, Huo HX, Li W, Li JD, et al. [L-carnitine: safe and effective for asthenozoospermia]. Zhonghua Nan Ke Xue. 2010;16(5):420-2.Chinese.
  27. Shi JZ, Zhang SS, Zhang Z, Liang Q, Shi Y, Hua JL, et al. [Expressions of sperm-specific genes in carnitine-cultured testis sperm of obstructive azoospermia patients]. Zhonghua Nan Ke Xue. 2010;16(6):504-9. Chinese.
  28. Jeulin C, Lewin LM. Role of free L-carnitine and acetyl-L-carnitine in post-gonadal maturation of mammalian spermatozoa. Hum Repord Update. 1996;2(2):87-102.
  29. Brooks DE. Carnitine, acetylcarnitine and the activity of carnitine acyltransferases in seminal plasma and spermatozoa of men, rams and rats. J Reprod Fertil. 1979;56(2):667-73.
  30. Casillas ER, Chaipayungpan S. The distribution of carnitine and acetylcarnitine in the rabbit epididymis and the carnitine content of rabbit spermatozoa during maturation. J Reprod Fertil. 1979;56(2):439-44.
  31. Burgos C, Maldonado C, Gerez de Burgos NM, Aoki A, Blanco A. Intracellular localization of the testicular and sperm-specific lactate dehydrogenase isozyme C4 in mice. Biol Reprod. 1995;53(1):84-92.
  32. Tanphaichitr N. In vitro stimulation of human sperm motility by acetylcarnitine. Int J Fertil. 1977;22(2):85-91.
  33. Tasdemir I, Taşdemir M, Tavukcuoglu S. Effect of pentoxifylline on immotile testicular spermatozoa. J Assist Reprod Genet. 1998;15(2):90-2.
Journal of Reproduction & Infertility
Volume 14, Issue 2
Spring 2013
Pages 56-61