Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

Abstract

The in vitro production and cryopreservation of mammalian embryos generates reactive oxygen species (ROS) due to conditions of the system that can overcome their antioxidant protection. Resveratrol is an antioxidant used in in vitro systems to improve blastocyst rates, but its effect on antioxidant enzymes such as glutathione (GSH) in embryos produced by in vitro fertilization (IVF) after vitrification has not been reported. The objective of this study was to evaluate the effects of resveratrol on the in vitro maturation medium (IVM) of sheep oocytes (Ovis aries) on the levels of ROS and GSH in embryos produced by IVF subjected to vitrification. Resveratrol was added at 0 µM, 0.25 µM, 0.5 µM, and 1 µM during oocyte in vitro maturation (IVM). Matured oocytes were fertilized with thawed ram sperm. Embryos were cultured in sequential media until blastocysts, were then vitrified for 24 h, and, after heating, they were stained with DCFH-DA (2′,7′-dichlorodihydrofluorescein diacetate) to determine the presence of ROS and with Cell Tracker Blue® for the presence of GSH. The quantitative values of ROS and GSH were obtained through the Image J image processor. The results showed that resveratrol increased GSH and decreased ROS production (p < 0.05) in a dose-dependent manner. It is concluded that its use in sheep oocytes during IVM has a beneficial effect on embryos produced by IVF subjected to vitrification by decreasing reactive oxygen species levels and facilitating the generation of embryo antioxidant enzymes like glutathione.

The walnut-derived peptide TW-7 improves mouse parthenogenetic embryo development of vitrified MII oocytes potentially by promoting histone lactylation

“…Oocyte vitrification is increasingly used for female fertility preservation. However, as previously described, vitrification may cause damage to the endogenous antioxidant system and redox homeostasis imbalance in several mammalian oocytes [ 57 – 65 ], which puts a great adverse impact on the developmental potential of the oocyte [ 21 , 58 – 60 ]. In the present study, we found that the PA zygotes derived from vitrified mouse MII oocytes showed the increased oxidative stress.…”

Effects of Different Diluents and Freezing Methods on Cryopreservation of Hu Ram Semen

“…The level of ROS production in the sperm was evaluated by a fluorescent probe with 2,7-dichlorodi-hydrofluorescein diacetate (DCFH-DA, Beyotime, Shanghai, China) [ 24 ]. Briefly, the thawed semen was diluted in basic diluent (1:6) and 50 µL of samples were mixed with DCFH-DA (10 mM).…”