Protective Effect of Cimicifuga racemosa (L.) Nutt Extract on Oocyte and Follicle Toxicity Induced by Doxorubicin during In Vitro Culture of Mice Ovaries

Abstract

This study evaluated the potential of Cimicifuga racemosa (L.) Nutt extract (CIMI) to reduce the deleterious effects of doxorubicin (DOXO) in oocytes, follicles and stromal cells in mice ovaries cultured in vitro. In experiment 1, mice ovaries were cultured in DMEM+ alone or supplemented with 5, 50 or 500 ng/mL CIMI, while in experiment 2, mice ovaries were cultured in DMEM+ alone or supplemented with 5 ng/mL CIMI (better concentration), 0.3 μg/mL DOXO or both. Thereafter, the ovaries were processed for histological (morphology, growth, activation, extracellular matrix configuration and stromal cell density), immunohistochemical (caspase-3) analyses. Follicle viability was evaluated by fluorescence microscopy (ethidium homodimer-1 and calcein) while real-time PCR was performed to analyses the levels of (mRNA for SOD, CAT and nuclear factor erythroid 2–related factor 2 (NRF2) analyses. The results showed that DOXO reduces the percentage of normal follicles and the density of stromal cells in cultured ovaries, but these harmful effects were blocked by CIMI. The DOXO reduced the percentage of primordial follicles, while the presence of CIMI alone did not influence percentage of primordial follicles. A higher staining for caspase-3 was seen in ovaries cultured in control medium alone or with DOXO when compared with those cultured with CIMI alone or both CIMI and DOXO. In addition, follicles from ovaries cultured with both CIMI and DOXO were stained by calcein, while those follicles cultured with only DOXO were stained with ethidium homodimer-1. Furthermore, ovaries cultured with CIMI or both CIMI and DOXO had higher levels of mRNA for SOD and CAT, respectively, than those cultured with only DOXO. In conclusion, the extract of CIMI protects the ovaries against deleterious effects of DOXO on follicular survival and ovarian stromal cells.

A medicina voltada à promoção da saúde e do bem-estar 4

“…racemosa (5 ng/mL) adicionada ao meio de cultivo foi capaz de proteger o tecido ovariano dos efeitos negativos da DOX, diminuindo a atresia folicular e auxiliando na manutenção da densidade celular do estroma ovariano (DE ASSIS et al, 2022). Nesse estudo, houve elevação da expressão de enzimas antioxidantes, além da diminuição da apoptose.…”

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone

“…Studies to investigate the potential of natural substances capable of attenuating the deleterious effects of GCs on ovarian function in order to preserve fertility are extremely necessary. Our recent results showed that Cimicifuga racemosa (L.) Nutt (CIMI) extract protects ovarian follicles from atresia induced by doxorubicin in cultured ovaries (12).…”

In Vitro Growth of Mammalian Follicles and Oocytes

“…de Assis et al [37] also focused on an antioxidant, Cimicifuga racemosa (L.) Nutt extract (CIMI) [38], to investigate its protective effects on mouse ovaries cultured with Doxorubicin, a drug for cancer treatment with gonadotoxic side effects. They showed that CIMI protected follicles from Doxorubicin-induced apoptosis by reducing oxidative stress, which might be applied for fertility preservation in patients with cancer.…”

“…In conclusion, this Special Issue covers culture systems from a wide range of follicular developmental stages, including primordial follicles [39], secondary follicles [35,39], early antral follicles [31], small antral follicles [41], in vivo grown antral follicles [35], and whole ovarian cultures [37]. Moreover, it consists of studies of four different species, including water buffaloes [31], cattle [33,35,39], mice [37], and pigs [39].…”

“…In conclusion, this Special Issue covers culture systems from a wide range of follicular developmental stages, including primordial follicles [39], secondary follicles [35,39], early antral follicles [31], small antral follicles [41], in vivo grown antral follicles [35], and whole ovarian cultures [37]. Moreover, it consists of studies of four different species, including water buffaloes [31], cattle [33,35,39], mice [37], and pigs [39]. This variety suggests the need to study oocytes from early developmental stages in varied fields, as well as the importance of the development and refinement of culture systems for each follicle's developmental stage.…”