N-乙酰半胱氨酸对不同直径卵泡来源猪卵母细胞体外成熟效果的影响

doi:10.11843/j.issn.0366-6964.2023.03.018

摘要/Abstract

摘要： 旨在在猪卵母细胞体外成熟培养液中添加N-乙酰半胱氨酸(NAC)，揭示NAC对不同直径卵泡来源卵母细胞体外成熟效果的影响，解析其对氧化还原平衡的调控作用。本研究收集猪卵巢上大腔(4~6 mm)、中腔(2~4 mm)、小腔(1~2 mm)卵泡中的卵母细胞，在以上3种卵泡来源猪卵母细胞的体外成熟培养液中，均分别添加0、1、2、4 mmol·L^-1浓度的NAC处理，评价卵丘扩展指数、第一极体排出率等成熟指标，检测卵母细胞中ROS水平、谷胱甘肽(GSH)含量和抗氧化基因的表达水平。结果表明，NAC处理对大腔卵泡来源卵母细胞的卵丘扩展指数和ROS水平无显著影响(P>0.05)，对大、中腔卵泡来源卵母细胞的第一极体排出率无显著作用(P>0.05)；而适量浓度的NAC(2 mmol·L^-1)处理可显著提高中、小腔卵泡来源卵母细胞的卵丘扩展指数(P<0.05)，降低ROS水平(P<0.05)，提升小腔卵泡来源卵母细胞的第一极体排出率(P<0.05)。同时，NAC处理对大、中腔卵泡来源卵母细胞中的GSH含量和抗氧化基因SOD、CAT、Prdx2、Prdx6的表达水平无显著影响(P>0.05)，而适量浓度的NAC(2 mmol·L^-1)处理可显著提高小腔卵泡来源卵母细胞中的GSH含量(P<0.05)，上调抗氧化基因SOD、CAT的表达水平(P<0.05)。综上所述，适当浓度的NAC(2 mmol·L^-1)处理可以通过缓解氧化应激提高小腔卵泡来源卵母细胞的体外成熟效果。

关键词: N-乙酰半胱氨酸, 不同直径卵泡, 猪卵母细胞, 体外成熟, 氧化应激

Abstract: In this study, N-acetylcysteine (NAC) was added into the culture medium of porcine oocytes for in vitro maturation to reveal the effects of NAC on the in vitro maturation of porcine oocytes from follicles with different sizes and to analyze the regulation of NAC on redox balance. Oocytes from large follicles (4-6 mm), middle follicles (2-4 mm) and small follicles (1-2 mm) in porcine ovary were collected and the three types of porcine oocytes were respectively cultured with 0, 1, 2, 4 mmol·L^-1 NAC. The maturation indexes such as cumulus expansion index and the first polar body excretion rate were evaluated, and ROS level, glutathione (GSH) content and expression level of antioxidant genes in oocytes were detected. The results showed that NAC treatment had no significant effect on cumulus expansion index and ROS level of oocytes from large follicles (P>0.05), and had no significant effect on the first polar body excretion rate of oocytes from large and middle follicles (P>0.05), however proper concentration of NAC (2 mmol·L^-1) significantly increased the cumulus expansion index and decreased ROS level (P<0.05) of oocytes from middle and small follicles (P<0.05), and significantly increased the first polar body excretion rate of oocytes from small follicles (P<0.05). Meanwhile, NAC treatment had no significant effect on GSH content and expression levels of antioxidant genes SOD, CAT, Prdx2 and Prdx6 of oocytes from large and middle follicles (P>0.05), and proper concentration of NAC (2 mmol·L^-1) significantly increased GSH content and up-regulated the expression levels of SOD and CAT of oocytes from small follicles (P<0.05). In conclusion, proper concentration of NAC (2 mmol·L^-1) can improve the in vitro maturation effect of oocytes from small follicles by alleviating oxidative stress.

Key words: N-acetylcysteine, follicles with different diameters, porcine oocyte, in vitro maturation, oxidative stress

中图分类号:

S828.3

肖十雨, 卢畅, 马娟, 王闯, 亓美玉, 姚玉昌. N-乙酰半胱氨酸对不同直径卵泡来源猪卵母细胞体外成熟效果的影响[J]. 畜牧兽医学报, 2023, 54(3): 1046-1057.

XIAO Shiyu, LU Chang, MA Juan, WANG Chuang, QI Meiyu, YAO Yuchang. Effects of N-acetylcysteine[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1046-1057.

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