Current Medical Science

, Volume 37, Issue 6, pp 915–921 | Cite as

Acetyl-L-carnitine: An effective antioxidant against cryo-damage on human spermatozoa with asthenospermia

  • Yu-jie Zou (邹宇洁)
  • Jing Yang (杨 菁)Email author
  • Shuo Chang (常 硕)
  • Wang-ming Xu (徐望明)
  • Tai-lang Yin (尹太郎)
  • Wen Long (龙 文)


A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen (ROS) level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations (group B: 2.5 mmol/L, group C: 7.5 mmol/L, group D: 15 mmol/L) was compared with control (group A: no acetyl-L-carnitine given). For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate (CR), tail DNA percentage (TD), tail length (TL) and Oliver tail moment (OTM) were significantly reduced in group C and group D. Group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.

Key words

acetyl-L-carnitine human spermatozoa DNA damage acrosome integrity 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Oehninger S, Duru NK, Srisombut C, et al. Assessment of sperm cryodamage and strategies to improve outcome. Mol Cell Endocrinol, 2000,169(1-2):3–10CrossRefPubMedGoogle Scholar
  2. 2.
    Hammitt DG, Bedia E, Rogers P, et al. Comparison of motility stimulants for cryopreserved human semen. Fertil Steril, 1989,52(3):495–502CrossRefPubMedGoogle Scholar
  3. 3.
    Wang R, Sikka SC, Veeraragavan K, et al. Platelet activating factor and pentoxifylline as human sperm cryoprotectants. Fertil Steril, 1993,60(4):711–715CrossRefPubMedGoogle Scholar
  4. 4.
    Ford WCL, Rees JM, McLaughlin EA, et al. Pentoxifylline acts synergistically with A23187 to increase the penetration of zona-free hamster oocytes by cryopreserved human spermatozoa. Int J Androl, 1994,17(4):199–204CrossRefPubMedGoogle Scholar
  5. 5.
    Sharma RX, Agarwal A. Artificial stimulation of cryopreserved human spermatozoa by sodium nitroprusside, 2-chloroadenosine, and 2-deoxyadenosine. Eur Urol, 1997,32(3):344–352PubMedGoogle Scholar
  6. 6.
    Sbracia M, Grasso J, Sayme N, et al. Hyaluronic acid substantially increases the retention of motility in cryopreserved-thawed human spermatozoa. Hum Reprod, 1997,12(9):1949–1954CrossRefPubMedGoogle Scholar
  7. 7.
    Siow Y, Fallat ME, Amin FA, et al. Mullerian inhibiting substance improves longevity of motility and viability of fresh and cryopreserved sperm. J Androl, 1998,19(5):568–572PubMedGoogle Scholar
  8. 8.
    Esteves SC, Sharma RK, Thomas AJ, et al. Cryopreservation of human spermatozoa with pentoxifylline improves the post-thaw agonist-induced acrosome reaction rate. Hum Reprod, 1998,13(12):3384–3389CrossRefPubMedGoogle Scholar
  9. 9.
    Sánchez-Calabuig MJ, Maillo V, Beltrán-Breña P, et al. Cryopreservation of canine sperm using egg yolk and soy bean based extenders. Reprod Biol, 2017,17(3):233–238CrossRefPubMedGoogle Scholar
  10. 10.
    Osipova VP, Berberova NT, Gazzaeva RA, et al. Application of new phenolic antioxidants for cryopreservation of sturgeon sperm. Cryobiology, 2016,72(2):112–118CrossRefPubMedGoogle Scholar
  11. 11.
    Trzcinska M, Bry1a M, Gajda B, et al. Fertility of boar semen cryopreserved in extender supplemented with butylated hydroxytoluene. Theriogenology, 2015,83(3):307–313CrossRefPubMedGoogle Scholar
  12. 12.
    Bucak MN, Sarıözkan S, Tuncer PB, et al. The effect of antioxidants on post-thawed Angora goat (Capra hircus ancryrensis) sperm parameters, lipid peroxidation and antioxidant activities. Small Ruminant Res, 2010,89(5):24–30CrossRefGoogle Scholar
  13. 13.
    Bucak MN, Tuncer PB, Sarıözkan S, et al. Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: Antioxidants protect DNA integrity against cryodamage. Cryobiology, 2012,61(1):248–253Google Scholar
  14. 14.
    Duru NK, Morshedi M, Schuffner A, et al. Semen treatment with progesterone and/or acetyl-L-carnitine does not improve sperm motility or membrane damage after cryopreservation-thawing. Fertil Steril, 2000,74(4):715–720CrossRefPubMedGoogle Scholar
  15. 15.
    World Health Organization. Laboratory manual for the examination of human semen and semen-cervical mucus interaction. 4th ed. Cambridge, United Kingdom: The Press Syndicate of the University of Cambridge, 1999.Google Scholar
  16. 16.
    Reid CE, Hermes R, Blottner S, et al. Split-sample comparison of directional and liquid nitrogen vapour freezing method on post-thaw semen quality in white rhinoceroses (Ceratotherium simum simum and Ceratotherium simum cottoni). Theriogenology, 2009,71(2):275–291CrossRefPubMedGoogle Scholar
  17. 17.
    Aramli MS, Golshahi K, Nazari RM, et al. Effect of freezing rate on motility, adenosine triphosphate content and fertilizability in beluga sturgeon (Huso huso) spermatozoa. Cryobiology, 2015,70(2):170–174CrossRefPubMedGoogle Scholar
  18. 18.
    Meamar M, Zribi N, Cambi M, et al. Sperm DNA fragmentation induced by cryopreservation: new insights and effect of a natural extract from Opuntia ficus-indica. Fertil Steril, 2012,98(2):326–333CrossRefPubMedGoogle Scholar
  19. 19.
    Cavallini G, Modenini F, Vitali G, et al. Acetyl-L-carnitine plus propionyl-L-carnitine improve efficacy of sildenafil in treatment of erectile dysfunction after bilateral nerve-sparing radical retropubic prostatectomy. Urology, 2005,66(5):1080–1085CrossRefPubMedGoogle Scholar
  20. 20.
    Lenzi A, Lombardo F, Sgro P. Use of carnitine therapy in selected cases of male factor infertility: a double-blind crossover trial. Fertil Steril, 2003,79(2):292–300CrossRefPubMedGoogle Scholar
  21. 21.
    Medeiro CMO, Forell F, Oliveira ATD, et al. Current status of sperm cryopreservation: Why isn’t it better? Theriogenology, 2002,57(1):327–344CrossRefGoogle Scholar
  22. 22.
    Balercia G, Regoli F, Armeni T, et al. Placebo-controlled double-blind randomized trial on the use of L-carnitine, L-acetylcarnitine, or combined L-carnitine and L-acetylcarnitine in men with idiopathic asthenozoospermia. Fertil Steril, 2005,84(3):662–671CrossRefPubMedGoogle Scholar
  23. 23.
    Mongioi L, Calogero AE, Vicari E, et al. The role of carnitine in male infertility. Andrology, 2016,4(5):800–807CrossRefPubMedGoogle Scholar
  24. 24.
    Ng CM, Blackman MR, Wang C, et al. The role of carnitine in the male reproductive system. Ann NY Acad Sci, 2004,1033:177–188CrossRefPubMedGoogle Scholar
  25. 25.
    Vicari E, La Vignera S, Calogero AE. Antioxidant treatment with carnitines is effective in infertile patients with prostatovesiculoepididymitis and elevated seminal leukocyte concentrations after treatment with nonsteroidal anti-inflammatory compounds. Fertil Steril, 2002,78(6):1203–1208CrossRefPubMedGoogle Scholar
  26. 26.
    Garolla A, Maiorini M, Roverato A, et al. Oral carnitine supplementation increases sperm motility in asthenozoospermic men with normal sperm phospholipid hydroperoxide glutathione peroxidase levels. Fertil Steril, 2005,83(2):662–671CrossRefGoogle Scholar
  27. 27.
    Zopfgen A, Priem F, Sudhoff F. Relationship between semen quality and the seminal plasma components carnitine, alpha-glucosidase, fructose, citrate and granulocyte elastase in infertile men compared with a normal population. Hum Reprod, 2000,15(4):840–845CrossRefPubMedGoogle Scholar
  28. 28.
    Gurbuz B, Yalti S, Ficicioglu C, et al. Relationship between semen quality and seminal plasma total carnitine in infertile men. J Obstet Gynaecol, 2003,23(6):653–656CrossRefPubMedGoogle Scholar
  29. 29.
    Li K, Li W, Huang YF. Determination of free L-carnitine in human seminal plasma by high performance liquid chromatography with pre-column ultraviolet derivatization and its clinical application in male infertility. Clinica Chimica Acta, 2007,378(1):159–163CrossRefGoogle Scholar
  30. 30.
    Partyka A, Rodak O, Bajzert J, et al. The effect of L-carnitine, hypotaurine, and taurine supplementation on the quality of cryopreserved chicken semen. Biomed Res Int, 2017,2017:7279341CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Gandini L, Lombardo F, Lenzi A, et al. Cryopreservation and sperm DNA integrity. Cell Tissue Bank, 2006,7(5):91–98CrossRefPubMedGoogle Scholar
  32. 32.
    Donnelly ET, Steele EK, McClure N, et al. Assessment of DNA integrity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation. Hum Reprod, 2001,16(6):1191–1199CrossRefPubMedGoogle Scholar
  33. 33.
    Kalthur G, Adiga SK, Upadhya D, et al. Effect of cryopreservation on sperm DNA integrity in patients with teratospermia. Fertil Steril, 2008,89(6):1723–1727CrossRefPubMedGoogle Scholar
  34. 34.
    Yan W. Male infertility caused by spermiogenic defects: Lessons from gene knockouts. Mol Cell Endocrinol, 2009,306(1-2):24–32CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Valcircel A, Heras de las MA, Pkrez L, et al. Assessment of the acrosomal status of membrane-intact ram spermatozoa after freezing and thawing by simultaneous lectin/Hoechst 33258 staining. Animal Reprod Sci, 1997,45(4):299–309CrossRefGoogle Scholar

Copyright information

© Huazhong University of Science and Technology 2017

Authors and Affiliations

  • Yu-jie Zou (邹宇洁)
    • 1
  • Jing Yang (杨 菁)
    • 1
    Email author
  • Shuo Chang (常 硕)
    • 1
  • Wang-ming Xu (徐望明)
    • 1
  • Tai-lang Yin (尹太郎)
    • 1
  • Wen Long (龙 文)
    • 1
  1. 1.Reproductive Medicine CenterRenmin Hospital of Wuhan UniversityWuhanChina

Personalised recommendations