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

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«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