OOCYTES SINGLE-STRAND DNA BREAKS WITH THE INFLUENCE OF ANTIOXIDANT, PARP-1 IHIBITOR, NO SYNTHASE SUBSTRATE, iNOS IHIBITOR

Sribna V. O., Grushka N. G., Kondratska O. A., Blashkiv T. V., Yanchiy R. I.

CLINICAL AND EXPERIMENTAL MEDICINE

Scentific article

Premature ovarian failure (POF) – an ovarian function disorder affecting women under 40 – is actively studied. This disease is fairly widespread due to the delay in maternal age, and today is a medical and social problem. POF is studied on models using rodents and main researches are aimed at correction of ovarian disorder. Previously, using a model of experimental immune complex-mediated failurе, which partially reflects POF, inhibition of oocytes meiotic maturation and increased DNA damage of follicular cells surrounding oocytes, thymic and lymph nodes cells (increase of single-strand breaks of DNA) was established, and it was shown that these investigated parameters are improved with the use of antioxidant, iNOS ihibitor, because of the probable reduction of manifestations of oxidative-nitrosatitative stress. The purpose of the work was to investigate the distribution of single-strand DNA breaks of oocyte nuclei with the influence of antioxidant, PARP inhibitor, iNO-synthase inhibitor, NOS substrate. The study was carried out on non-pregnant female BCA mice. For DNA damage assessment DNA-comet assay (alkaline) was used. Comets were separated into 4 classes (0/1; 2; 3; 4) depending on the value of DNA in the “head” and “tail” of comet. It was established that the PARP inhibitor – 4-Hydroxyquinazoline (4-GQ) and iNOS ihibitor – aminoguanidine (AG), as well as their combination, lead to the redistribution of single-strand DNA breaks of oocytes in the direction of increasing the DNA damage. NOS substrate L-arginine and antioxidant resveratrol did not have a damaging effect on the DNA of oocytes, on the contrary, there was a certain increase in the number of cells with nuclei 0/1 classe. It was obtained that under the influence of 4-GQ and L-arginine (1.0 mM / 0.04 mM, respectively) there was a decrease in the oocyte DNA damage in relation to the values in the group of 4-GQ: the particles of 3 and 4 classe`s nuclei decrease in, respectively, 1,7 and 2,1 times, and nuclei 0/1 and 2 classes increase in, respectively, 3,1 and 1,9 times; under the influence of 4-GQ and resveratrol (1.0 mM / 2.0μM, respectively), there was obtained a decrease in the oocyte DNA damage, namely, the proportion of cells with 3 and 4 class`s nuclei decrease in, respectively, 1,4 and 1,8 times, and the particles of nuclei 0/1, 2 classes increase in, respectively, 2,6 and 1,7 times as compared to those under the influence of 4-GQ. Under the influence of AG and resveratrol (0,02 mM / 2,0 μM, respectively), there was a decrease in the oocyte DNA damage, namely, the number of cells with 3 and 4 class nuclei was reduced to 22.78 ± 1.53% (p < 0.05, n = 6) and 25.19 ± 1.67% (p <0.05, n = 6) compared with, respectively, 32.96 ± 1.67% and 35.93 ± 1.67% under the influence of AG, and 0/1 class nuclei increased to 31.85 ± 1.67% (p <0.05, n = 6) relative to the value under the influence of AG – 12.22 ± 2.72%. Conclusions. The results of the study of the integral integrity of the oocyte genome using antioxidant (resveratrol), the PAPP-1 (4-GK) inhibitor, the NOS substrate (L-arginine), the iNOS inhibitor (aminoguanidine) give grounds to defend the NO-dependence of the DNA repair regulation, which is displayed as single-strand DNA breaks.

single-strand DNA breaks, oocytes, premature ovarian failure

• Kas’yan Ye.S. Problema prezhdevremennoy yaichnikovoy nedostatochnosti. Sovremennyye podkhody k diagnostike i lecheniyu / Ye.S. Kas’yan // Slovo o zdorov’ye. – 2017. – № 11. – S. 35-40.
• Makogon N.V. Polí(ADF-ribozo) polímeraza 1: fízíologíchna ta patofízíologíchna rol’ / N.V. Makogon, Í.M. Alekseyeva // Fízíologíchniy zhurnal – 2012. – T. 58, № 3. – S. 95-112.
• Sríbna V.O. Odnonitkoví rozrivi DNK yader klítin folíkulyarnogo otochennya ootsitív, timusa í límfatichnikh vuzlív za umov yeksperimental’nogo ímunnogo ushkodzhennya ta zastosuvannya antioksidanta / V.O. Sríbna, N.G. Grushka, R.Í. Yanchíy // Vísnik problem bíologíí̈ í meditsini. – 2016. – № 2, tom 3 (130). – S. 195-199.
• Funktsional’noye sostoyaniye yaichnika, matki, timusa i limfaticheskikh uzlov u myshey s eksperimental’nym immunokompleksnym povrezhdeniyem v usloviyakh vvedeniya substantsii nanochastits nol’-valentnogo zheleza / V.A. Sribnaya, A.P. Litvinenko, L.S. Reznichenko [ta ín.] // Problemy reproduktsii. – 2016. – № 22 (4). – S. 20-27.
• A review on PARP1 inhibitors: Pharmacophore modeling, virtual and biological screening studies to identify novel PARP1 inhibitors / S. Singh, J. Sarma, L. Narasu [et al.] // Curr Top Med Chem. – 2014. – № 14 (17). – Р. 2020-2030.
• Autoantibodies to heat-shock protein, HSPA5, and epitope spreading: high-dose dexamethasone therapy rescues ovarian function in experimental autoimmune ovarian insufficiency mouse model / K. Kadam, P. Mande, N. Gawas [et al.] // Am J Reprod Immunol. – 2016. – № 75 (5). – P. 580-593.
• Autoimmune polyglandular syndrome type 3 (APS-3) among patients with premature ovarian insufficiency (POI) / K. Szlendak- Sauer, D. Jakubik, M. Kunicki [et al.] // Eur J Obstet Gynecol Reprod Biol. – 2016. – № 203. – P. 61-65.
• Autoimmune primary ovarian insufficiency / C. Silva, L. Yamakami, N. Aikawa [et al.] // Autoimmun Rev. – 2014. – № 13 (4-5). – P. 427-430.
• Assessment of 1,2-propanediol (PrOH) genotoxicity on mouse oocytes by comet assay / A. Berthelot-Ricou, J. Perrin, C. di Giorgio [et al.] // Fertil Steril. – 2011. – Vol. 96. – Р. 1002-1007.
• Bukovsky A. Immunoregulation of follicular renewal, selection, POF, and menopause in vivo, vs. neo oogenesis in vitro, POF and ovarian infertility treatment, and a clinical trial / A. Bukovsky, M. Caudle // Reprod Biol Endocrinol. – 2012. – № 10. – P. 97-112.
• Growth hormone treatment of premature ovarian failure in a mouse model via stimulation of the Notch-1 signaling pathway / T. Liu, S. Wang, L. Zhang [et al.] // Exp Ther Med. – 2016. – № 12 (1). – P. 215-221.
• Hydrogen-rich Water Exerting a Protective Effect on Ovarian Reserve Function in a Mouse Model of Immune Premature Ovarian Failure Induced by Zona Pellucida3 / X. He, S. Wang, C. Yin [et al.] // Chin Med J. – 2016. – № 129 (19). – P. 2331-2337.
• Psychosocial vulnerability, resilience resources, and coping with infertility: a longitudinal model of adjustment to primary ovarian insufficiency / M. Driscoll, M. Davis, L. Aiken [et al.] // Ann Behav Med. – 2016. – № 50 (2). – P. 272-284.
• Resveratrol during in vitro maturation improves the quality of bovine oocyte and enhances embryonic development in vitro / V. Torres, L. Muñoz, R. Urrego, [et al.] // Reprod Fertil Dev. – 2016. – № 29 (1). – P. 199-203.
• Resveratrol protects mouse oocytes from methylglyoxal-induced oxidative damage / Y. Liu, X. He, X. Huang [et al.] // PLoS One. – 2013. – № 8 (10). – P. 779-806.
• Resveratrol-induced mitochondrial synthesis and autophagy in oocytes derived from early antral follicles of aged cows / M. Sugiyama, R. Kawahara-Miki, H. Kawana [et al.] // J Reprod Dev. – 2015. – № 61 (4). – P. 251-259.
  17. Veith S. RecQ helicases and PARP1 team up in maintaining genome integrity / S. Veith, A. Mangerich // Ageing Res Rev. – 2015.
  – № 23 (A). – P. 12-28.

«Bulletin of problems biology and medicine» Issue 4 Part 3 (141), 2017 year, 231-234 pages, index UDK 616-092:618.112:618.112:616-092.4

10.29254/2077–4214–2017–4–3–141–231-234