Metformina, fisetina e resveratrol: possíveis protetores da reserva ovariana de camundongas expostas à ciclofosfamida

Abstract

Germ cells suffer irreversible damage during treatment with cytotoxic drugs such as cyclophosphamide (CYP). Its use drastically reduces female fertility, as this drug induces increased recruitment of quiescent follicles, anticipating menopause. CYP promotes various cellular damages that favor cellular senescence. This work is the first study evaluating strategies to prevent CYP-induced ovarian senescence through the administration of resveratrol (RES), fisetin (FIS) and metformin (MET). It is also the first experiment to elucidate the effect of these three compounds in order to preserve ovarian reserve in CYP-exposed mice. Sixty 60-day-old C57BL/6 females were used and divided into six experimental groups, one control (CTL) without any intervention, and five groups receiving CYP. Of these, one received no additional treatment and the other three groups received treatments with MET, FIS and RES alone while the last group received the combination of FIS+RES. These animals were euthanized after two weeks and the ovaries were collected for analysis. The primordial follicle reserve in the CYP group was significantly lower (p≤0.05) than in the CTL group. We observed no statistical difference with respect to total follicles. Although gene expression reflected inconclusive results, we observed significant reduction in p16 expression in the combination group (FIS+ RES+CYP; p≤0.05). There was higher lipofuscin labeling in the CYP group than in the other groups (p≤0.05). Therefore, we observed that CYP caused cellular damage that promoted greater senescence in the ovaries of the group that received CYP alone, thus demonstrating that the proposed interventions prevented damage that would result in the formation of senescent cells. Ovarian reserve was affected by CYP, however, the treatments showed only partial protective effect.