RESUMEN
This study aims to evaluate the effects of N-acetylcysteine (NAC) on bovine oocyte maturation, mitochondrial activity and transzonal projections (TZP), as well as on the levels of reactive oxygen species (ROS) and messenger RNA (mRNA) for catalase (CAT) superoxide dismutase (SOD), periredoxin-6 (Prdx6), glutathione peroxidase (GPx), growth and differentiation factor-9 (GDF9), histone H1Foo, cyclin B1 (CCNB1) and c-Mos. Bovine cumulus-oocyte complexes (COC) of medium-sized antral follicles (3.0-6.0 mm) were prematured in TCM-199 for 8 h at 38.5°C in 5% CO2. After prematuration in the presence of forskolin and C-type natriuretic peptide, COCs were matured in TCM-199 alone or with 0.1, 0.5 or 2.5 mM NAC. Then, oocytes were classified according to the stage of chromatin. Furthermore, mitochondrial activity and intracellular levels of ROS and TZP were also evaluated. The levels of mRNAs for CAT, SOD, Prdx6, GPx, GDF9, H1Foo, CCNB1 and c-Mos were evaluated using real-time polymerase chain reaction (RT-PCR). The results showed that NAC significantly increased the percentages of oocytes with resumption of meiosis when compared with those oocytes matured in control medium. Oocytes had homogeneous mitochondrial distribution, and those cultured with 0.1 and 0.5 mM NAC had lower levels of ROS when compared with the control. In addition, 0.5 mM NAC reduced TZP and the levels of mRNA for CCNB1. In contrast, NAC did not influence the expression of CAT, GPx, Prdx6, SOD, GDF9, H1Foo, and c-Mos. In conclusion, 0.5 mM NAC reduced the levels of ROS, TZP and mRNA for CCNB1, and improved in vitro resumption of meiosis in oocytes from medium-sized bovine antral follicles.
Asunto(s)
Acetilcisteína , Técnicas de Maduración In Vitro de los Oocitos , Bovinos , Animales , Técnicas de Maduración In Vitro de los Oocitos/métodos , Especies Reactivas de Oxígeno/metabolismo , Acetilcisteína/farmacología , Acetilcisteína/metabolismo , Oocitos , Meiosis , Superóxido Dismutasa/metabolismo , Glutatión Peroxidasa/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Oxidative stress is generated by an imbalance between reactive oxygen species (ROS) formation and cellular defense mechanisms. To reduce cellular damage caused by ROS in vivo or in vitro, N-acetyl-cysteine (NAC) is converted into metabolites that have the capacity of stimulating synthesis of glutathione (GSH) which functions directly as free radical scavengers. The NAC antioxidant potential evaluated to the greatest extent is the indirect action of NAC, as a precursor of GSH, with glutathione being the primary antioxidant in cells. During long-term preantral follicle culture, NAC has a synergic action with FSH and an important function in sustaining preantral follicle growth and follicle-cell viability in vitro. The NAC inclusion in in vitro maturation medium for cumulus-oocyte complexes (COC) leads to protection of oocytes from damage induced by heat stress, reductions in ROS, and increases in cumulus cell expansion. Developing embryos are susceptable to oxidative stress because of susceptability to cellular structure damage and not having well-developed defense mechanisms. Results from various indicate there are beneficial effects of NAC on embryonic development by increasing GSH biosynthesis and regulating cell proliferation. In addition, NAC is also an effective antioxidant during cryopreservation of ovarian follicles, oocytes and embryos, because inclusion of NAC in preservation medium leads to improvements in mitochondrial function and cell viability, and reductions in ROS and cellular apoptosis. In this review, there is evaluation of mechanisms of action of NAC and beneficial effects during in vitro culture of preantral follicles, as well as oocyte maturation, embryonic development and cryopreservation.