活性氧对猪卵母细胞SUMO-1表达及其质量的影响

doi:10.16431/j.cnki.1671-7236.2020.08.023

Abstract

Abstract: The purpose of this study was to investigate the effects of reactive hydrogen peroxide (H₂O₂) on the expression of the protein ubiquitin SUMO-1 and the ability of sperm-oocyte binding of porcine oocyte during in vitro maturation (IVM).The test was divided into 5 groups:0 (control),10,50,75 and 100 μg/mL of H₂O₂ treatment group.Western blotting,flow cytometry,Real-time quantiative PCR,Hoechst staining and other methods were used to detect porcine oocyte in vitro maturation,content of protein SUMOylation SUMO-1,cell viability,expression of apoptotic gene mRNA,solubility of zona pellucida (ZP) and sperm-oocyte binding.The results showed that 75 μg/mL H₂O₂ group compared with the control,10 and 50 μg/mL H₂O₂ groups,the in vitro maturation rate and cell viability of oocytes decreased significantly,while 75 μg/mL H₂O₂ group compared with other H₂O₂ groups,the ZP lysis time prolonged significantly,and the number of sperm adhering to the ZP of mature oocytes was reduced (P<0.05).SUMO-1 protein markers appeared at 77 and 18 ku,and the content of SUMO-1 protein was significantly reduced in 75 μg/mL H₂O₂ group when compare with the control,10 and 50 μg/mL H₂O₂ groups (P<0.05).75 μg/mL H₂O₂ group compared with the control,10 and 50 μg/mL H₂O₂ groups,Bcl-2 gene was significantly down-regulated (P<0.05),while 50 μg/mL H₂O₂ significantly up-regulated Bax gene level when compared with the control and 10 μg/mL H₂O₂ groups (P<0.05).In summary,H₂O₂ could regulate the SUMOylation level and sperm-oocyte binding ability of porcine oocytes during in vitro maturation.

Key words: reactive oxygen species (ROS); hydrogen peroxide (H₂O₂); oocyte; SUMOylation; cell viability; porcine

CLC Number:

S828.9

XU Da, LUO Xiaotong, CHEN Xuan, LYU Yanqiu, JIN Yi. Effects of ROS on Expression of SUMO-1 and Quality of Porcine Oocytes[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2544-2552.

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Effects of ROS on Expression of SUMO-1 and Quality of Porcine Oocytes

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