Protective Effect of <i>Cimicifuga racemosa</i> (L.) Nutt Extract on Oocyte and Follicle Toxicity Induced by Doxorubicin during In Vitro Culture of Mice Ovaries
Ernando I. T. de Assis,
Venância A. N. Azevedo,
Miguel F. De Lima Neto,
Francisco C. Costa,
Laís R. F. M. Paulino,
Pedro A. A. Barroso,
Mariana A. M. Donato,
Christina A. Peixoto,
Alane P. O. Do Monte,
Maria H. T. Matos,
Alana N. Godinho,
Jordânia M. O. Freire,
Ana L. P. S. Batista,
José R. V. Silva,
Anderson W. B. Silva
Affiliations
Ernando I. T. de Assis
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Venância A. N. Azevedo
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Miguel F. De Lima Neto
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Francisco C. Costa
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Laís R. F. M. Paulino
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Pedro A. A. Barroso
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Mariana A. M. Donato
Laboratory of Ultrastructure, CNPqAM/FIOCRUZ, Federal University of Pernambuco, Recife 50670-901, PE, Brazil
Christina A. Peixoto
Laboratory of Ultrastructure, CNPqAM/FIOCRUZ, Federal University of Pernambuco, Recife 50670-901, PE, Brazil
Alane P. O. Do Monte
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of São Francisco Valley, Rodovia BR 407, Km 12, Lote 543—Projeto de Irrigação Nilo Coelho—S/N, Petrolina 56300-990, PE, Brazil
Maria H. T. Matos
Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of São Francisco Valley, Rodovia BR 407, Km 12, Lote 543—Projeto de Irrigação Nilo Coelho—S/N, Petrolina 56300-990, PE, Brazil
Alana N. Godinho
Nucleus of Research in Animal Experimentation (NUPEX), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Jordânia M. O. Freire
Nucleus of Research in Animal Experimentation (NUPEX), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Ana L. P. S. Batista
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
José R. V. Silva
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
Anderson W. B. Silva
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Maurocélio Rocha Ponte 100, Sobral 62041-040, CE, Brazil
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.
