The protective effect of cilostazol on experimental ischemia/reperfusion injury in rats ovaries on in vitro fertilization outcomes

Abstract

Objective: Ovarian torsion decreases ovarian reserve because of ischemic and reperfusion damage it causes. In this study, we investigated the protective effect of cilostazol (CIL) on experimental ischemia (I) and ischemic-reperfusion (I/R) damage in rat ovaries with in vitro fertilization (IVF) results.

Materials and methods: Forty-eight adult female Sprague-Dawley albino rats were randomly assigned to 6 groups with 8 animals in each group: Sham (S), I, I/R, S + CIL, I + CIL and I/R + CIL. The I groups were subjected to bilateral adnexal torsion for 3 h, while the I/R and I/R + CIL groups received subsequent detorsion for 3 h. Twenty-two mg/kg of CIL was given via oral gavage 30 min before surgery on the I (I+ CIL) or reperfusion (I/R + CIL) groups. Oocytes were collected before the IVF procedure and after ovulation induction with 150-300 IU/kg pregnant mare serum gonadotropin.

Results: The metaphase oocytes reached their highest value of 4.73±0.96 in the S+ CIL group and reached their lowest value of 0.51±0.55 in the I/R group. There were statistically significant differences in the number of second-day embryos among the I, I+ CIL, and I/R and I/R+ CIL groups (p=0.000). When the groups were compared in terms of Anti-Müllerian hormone change, the highest decrease was observed in the I and I/R groups.

Conclusion: CIL pretreatment before surgery has a protective effect against I and I/R in rats with ovarian torsion.

Keywords: Reperfusion injuries; cilostazol; in vitro fertilization; ovarian torsion.

Figure 1

A comparison of the study groups’ total oocyte count, germinal vesicle (GV), metaphase I (MI), metaphase II (MII) oocyte counts, and second-day embryo counts. Statistical analysis between all groups was performed with a Kruskal-Wallis test. Pairwise comparisons were made with the Mann-Whitney test. The black cylinder indicates the statistically significant difference between the sham (S) and S+ (cilostazol) CIL groups. There was a statistically significant difference between the black star, triangle, circle and plus sign ischemia (I) and I + CIL groups. There was a statistically significant difference between the black pentegon, cross, equalitiy and rectangle ischemia reperfusion (I/R) and I/R + CIL groups

Figure 2

Comparison of second day embryos between groups. Second-day embryos of the groups are seen at 100 x magnification. The second-day embryo counts and embryo quality in the sham (S) and S+ cilostazol (CIL) groups were quite good compared to the other groups. It is seen that the quality and number of embryos on the second day are significantly better in the Ischemia (I) + CIL and ischemia reperfusion (I/R) + CIL groups compared to the I and I/R groups, respectively

Figure 3

Comparison of Anti-Müllerian hormone (AMH) levels after cilostazol administration between groups via an AMH1 and AMH2 correlation plot between the sham (S) + cilostazol (CIL), Ischemia (I) + CIL and Ischemia reperfusion (I/R) + CIL groups (mean and 95% confidence interval). There was no correlation between AMH1 and AMH2 values in the S + CIL group (P>0.05). A high level of correlation was observed between AMH1 and AMH2 values in the I + SI and I/R + SI groups. Analysis was done with a paired sample test. P<0.05 was considered statistically significant