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Previously it was reported that melatonin could mitigate oxidative stress caused by oocyte cryopreservation; however, the underlying molecular mechanisms which cause this remain unclear. The objective was to explore whether melatonin could reduce oxidative stress during in vitro maturation of vitrified-warmed mouse germinal vesicle (GV) oocytes through the Nrf2 signaling pathway or its receptors. During in vitro maturation of vitrified-warmed mouse GV oocytes, there were decreases ( p &lt; 0.05) in the development rates of metaphase I (MI) oocytes and metaphase II (MII) and spindle morphology grades; increases ( p &lt; 0.05) in the reactive oxygen species (ROS) levels; and decreases ( p &lt; 0.05) in expressions of Nrf2 signaling pathway-related genes ( Nrf2 , SOD1 ) and proteins (Nrf2, HO-1). However, adding 10 -7 mol/L melatonin to both the warming solution and maturation solutions improved ( p &lt; 0.05) these indicators. When the Nrf2 protein was specifically inhibited by Brusatol, melatonin did not increase development rates, spindle morphology grades, genes, or protein expressions, nor did it reduce vitrification-induced intracellular oxidative stress in GV oocytes during in vitro maturation. In addition, when melatonin receptors were inhibited by luzindole, the ability of melatonin to scavenge intracellular ROS was decreased, and the expressions of genes ( Nrf2 , SOD1 ) and proteins (Nrf2, HO-1) were not restored to control levels. Therefore, we concluded that 10 -7 mol/L melatonin acted on the Nrf2 signaling pathway through its receptors to regulate the expression of genes ( Nrf2 , SOD1 ) and proteins (Nrf2, HO-1), and mitigate intracellular oxidative stress, thereby enhancing in vitro development of vitrified-warmed mouse GV oocytes.

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Melatonin Promotes In Vitro Maturation of Vitrified-Warmed Mouse Germinal Vesicle Oocytes, Potentially by Reducing Oxidative Stress through the Nrf2 Pathway