RESUMO
Despite the cytotoxicity and embryotoxicity previously reported artesunate is a recommended drug to treat malaria for adults, children, and women in the first trimester of pregnancy. To address the putative effects of artesunate on female fertility and preimplantation embryo development, when the pregnancy is not detectable yet, artesunate was added to the oocyte in vitro maturation and in vitro embryo development of bovine. Briefly, in experiment 1 the cumulus-oocyte complexes (COCs) were in vitro matured for 18 h with 0.5, 1, or 2 µg/mL of artesunate or not (negative control) and then checked for nuclear maturation and subsequent embryo development. In experiment 2, the COCs were in vitro matured and fertilized without artesunate, which was added (0.5, 1, or 2 µg/mL) from the 1st to the 7th day of embryo culture along with a negative and a positive control group with doxorubicin. As a result, the use of artesunate on oocyte in vitro maturation did not differ from the negative control (p > 0.05) regarding nuclear maturation, cleavage, and blastocyst formation. Also, artesunate on in vitro embryo culture did not differ from negative control (p > 0.05) regarding cleavage and blastocyst formation, except for positive control, with doxorubicin (p < 0.05). In conclusion, under the conditions investigated, there was no evidence of artesunate toxicity on oocyte competence and the preimplantation period of in vitro embryo development in the bovine model, however, artesunate use still should be taken carefully as the outcome of implantation after oocytes and blastocysts exposure to artesunate remains unknown.
Artesunate added to in vitro maturation did not impair oocyte competence in bovine.Artesunate added to in vitro culture did not affect cleavage and blastocyst formation.No evidence of artesunate toxicity in oocytes and embryos of bovine was found.
RESUMO
Consequences of oocyte supplementation with l-carnitine may vary depending on species-specific cellular lipid profile, level of mitochondrial activity, or even on ipid availability in culture medium. This study aimed to evaluate l-carnitine supplementation on competence and gene expression of enzymes related to lipid metabolism in oocytes and cumulus cells from buffalo COCs matured in the presence or absence of fetal bovine serum (FBS). COCs were matured in vitro in FBS (10%) or bovine serum albumin fatty acid-free (BSA-FAF) (0.4%) and with or without supplementation with l-carnitine (3.03 mM). COCs matured in the presence of FBS or BSA-FAF were fertilized and cultured, then supplemented with l-carnitine during in vitro maturation or in vitro embryo culture. Finally, in vivo mature and immature COCs were included for gene expression analysis. COCs matured in culture medium with FBS in the presence of l-carnitine produced a lower blastocyst rate (p ≤ 0.05) compared to controls. In turn, the blastocyst rate from COCs matured with BSA-FAF in the presence of l-carnitine was similar to controls (p > 0.05), and higher than FBS + L-carnitine treated COCs (p ≤ 0.05). Addition of l-carnitine during embryo culture showed no differences in blastocyst production between experimental groups and controls (p > 0.05). In cumulus cells, gene expression of ACACA, SCD and FASN was upregulated in COCs matured in the presence of BSA-FAF + L-carnitine, while all genes in oocytes were significantly expressed upregulated by COCs matured in vivo, and only BSA-FAF + L-carnitine group showed similar expression of the FASN gene. In conclusion, the consequences of l-carnitine supplementation during in vitro maturation of buffalo COCs on oocyte competence vary depending on presence or absence of FBS in culture. With FBS, l-carnitine impairs oocyte competence, while in its absence, gene expression suggests adequate lipid metabolism and increased oocyte competence.
