Ukr.Biochem.J. 2021; Volume 93, Issue 3, May-Jun, pp. 24-29
doi: https://doi.org/10.15407/ubj93.03.024
Lipid peroxidation and DNA fragmentation in fresh and cryopreserved spermatozoa of men at different spermatogenesis state
T. O. Yurchuk*, O. V. Pavlovich, G. O. Gapon,
A. Y. Pugovkin, M. P. Petrushko
Institute for Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Department of Cryobiology of Reproductive System, Kharkiv;
*e-mail: taisiya.yur@gmail.com
Received: 11 July 2021; Accepted: 17 May 2021
Cryopreservation of spermatozoa is widely used in the treatment of infertility by assisted reproductive technologies. However, the cryopreservation causes an oxidative stress which can induce pathological changes in the male gametes. The aim of the research was to evaluate lipid peroxidation (LPO) and DNA fragmentation as well as correlation between these parameters in the fresh and cryopreserved spermatozoa of men with normozoospermia or oligoasthenoteratozoospermia (OAT) and spermatozoa derived from the epididymis of men with azoospermia. The level of malondialdehyde (MDA) in a TBA test, superoxide dismutase (SOD) activity and total antioxidant activity (AOA) were assessed. DNA integrity was estimated by acridine orange staining technique. It was shown that MDA level and SOD activity were significantly higher in the fresh spermatozoa of oligoasthenoteratozoospermic group compared with the fresh spermatozoa of normozoospermic group. After cryopreservation the MDA level increased in all groups and was the highest in OAT group where the greatest AOA decline was detected. DNA fragmentation frequency was 2.6 and 4.1 times higher in the fresh and cryopreserved OAT spermatozoa respectively as compared with the fresh normozoospermic ones (7.2%). DNA fragmentation was found to be the lowest in the fresh (6.2%) and cryopreserved (5.8%) epididymal spermatozoa. After cryopreservation SOD activity in epididymal spermatozoa was lower than in normozoaspermic. In spermatozoa of the studied groups the MDA level positively correlated with DNA fragmentation (0.79 Pearson’s correlation coefficient) both before and after cryopreservation. It is suggested that due to the detected low DNA fragmentation and LPO level in epididymal spermatozoa their use could be an alternative approach for infertility treatment by assisted reproductive technologies.
Keywords: antioxidant activity lipid peroxidation, cryopreservation, different spermatogenesis state, DNA fragmentation, MDA, SOD, spermatozoa
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