添加罗格列酮对小鼠卵母细胞氧化还原稳态的影响

doi:10.16431/j.cnki.1671-7236.2023.05.009

摘要/Abstract

摘要： 【目的】探究在体外成熟培养基中添加罗格列酮(rosiglitazone,RSG)对小鼠卵母细胞体外成熟及胚胎发育的影响,为体外成熟培养系统的优化提供参考。【方法】通过在体外成熟培养基中添加0、10、20、50、100 μmol/L RSG,统计第一极体(PBⅠ)排出率,确定RSG最适浓度。根据RSG最适浓度筛选结果分为对照组和罗格列酮组,通过免疫荧光染色法检测卵母细胞内活性氧(ROS)、谷胱甘肽(GSH)、组织蛋白酶B(CB)、线粒体膜电位水平(MMP)及囊胚内总细胞数(Hoechst 33342)和细胞凋亡比率(TUNNEL);使用实时荧光定量PCR检测囊胚内抗氧化相关基因(Sod-2、Gpx-3和Cat)和凋亡相关基因(Caspase-3、Bcl-2和Bcl-xl)转录水平。【结果】与对照组相比,添加20 μmol/L RSG可显著提高小鼠卵母细胞第一极体排出率、卵裂率及囊胚率(P＜0.05);卵母细胞内ROS和CB水平均显著降低,GSH和线粒体膜电位水平均显著提高(P＜0.05);囊胚内总细胞数显著提高,细胞凋亡比率显著降低(P＜0.05);抗氧化相关基因(Sod-2、Gpx-3和Cat)和抗凋亡相关基因(Bcl-2和Bcl-xl)转录水平显著提高,促凋亡基因Caspase-3转录水平显著降低(P＜0.05)。【结论】在体外成熟培养基中添加RSG对卵母细胞内氧化还原稳态有积极作用,可改善线粒体功能,提高卵母细胞体外受精后发育能力,并降低卵母细胞内CB水平及囊胚内细胞凋亡的发生。

关键词: 罗格列酮(RSG); 小鼠; 卵母细胞; 体外成熟(IVM); 体外受精(IVF)

Abstract: 【Objective】 This study was aimed to investigate the effects of rosiglitazone (RSG) on the oocyte maturation in vitro and subsequent development potential of embryos in mice,in order to provide reference for the optimization of in vitro maturation (IVM) culture system.【Method】 The optimum concentration of RSG was determined by adding 0,10,20,50 and 100 μmol/L RSG to IVM medium,and calculating the first polar body (PBⅠ) rate. According to the screening results of the RSG optimal concentration,they were divided into control and rosiglitazone groups.Reactive oxygen species (ROS),glutathione (GSH),cathepsin B (CB),mitochondrial membrane potential (MMP),total cell number (Hoechst 33342) and apoptosis ratio (TUNNEL) in oocytes or blastocysts were detected by immunofluorescence staining.The transcription levels of antioxidant genes (Sod-2,Gpx-3 and Cat) and apoptosis-related genes (Caspase-3,Bcl-2 and Bcl-xl) in blastocysts were detected by Real-time quantitative PCR.【Result】 Compared with control group,20 μmol/L RSG supplementation could significantly increase the PBⅠ rate,cleavage rate and blastocyst rate of oocytes in mice (P＜0.05);The intracellular ROS and CB levels were significantly decreased,the intracellular GSH and mitochondrial membrane potential levels were significantly increased (P＜0.05);The total cell number in blastocysts was significantly increased,and the apoptosis rate in blastocysts was significantly decreased (P＜0.05).The transcription levels of antioxidant genes (Sod-2, Gpx-3 and Cat) and anti-apoptotic genes (Bcl-2 and Bcl-xl) were significantly increased, and the transcription level of apoptotic gene Caspase-3 was significantly decreased (P＜0.05).【Conclusion】 The addition of RSG to IVM medium was beneficial to intracellular redox homeostasis of oocytes,which could improve the mitochondrial function,increase the developmental ability of oocytes after IVF,and decrease the intracellular CB level and apoptosis of cells in blastocysts.

Key words: rosiglitazone (RSG); mice; oocyte; in vitro maturation (IVM); in vitro fertilization (IVF)

中图分类号:

S852.1

张笑梦, 焦安惠, 王玉琪, 方南洙, 金庆国. 添加罗格列酮对小鼠卵母细胞氧化还原稳态的影响[J]. 中国畜牧兽医, 2023, 50(5): 1818-1827.

ZHANG Xiaomeng, JIAO Anhui, WANG Yuqi, FAN Nanzhu, JIN Qingguo. Effects of Rosiglitazone Supplementation on Redox Homeostasis of Oocytes in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1818-1827.

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