bFGF对3-硝基丙酸处理的小鼠受精卵体外发育能力的影响

doi:10.16431/j.cnki.1671-7236.2022.02.022

Abstract

Abstract: 【Objective】 This experiment was to study the effect of adding basic fibroblast growth factor (bFGF) to the in vitro development medium of mouse fertilized eggs treated with 3-nitropropionic acid (3-NPA), in order to provide reference for improving the in vitro development quality of early embryos under oxidative stress.【Method】 0, 20, 50, 100 and 150 ng/mL bFGF were added to the in vitro culture medium of mouse zygotes, cultured for 24, 48 and 96 h, and the 2-cell rate, 4-cell rate and blastocyst rate were counted to screen the best bFGF treatment concentration.In vivo zygotes were produced by intraperitoneal injection of 12.5 mg/kg 3-NPA, and the same volume of normal saline was injected intraperitoneally as the control, the fertilized eggs were divided into 3-NPA and 3-NPA+bFGF groups, control group (C) and bFGF group.After blastocysts were obtained, the level of reactive oxygen species (ROS) in embryos was detected by DCFH-DA, the level of glutathione (GSH) was detected by CMF2HC, and the mitochondrial membrane potential intensity of early embryos was detected by JC-1.【Results】 There was no significant difference in 2-cell rate among 0, 20, 50, 100 and 150 ng/mL bFGF groups (P > 0.05), and the blastocyst rate in 100 ng/mL bFGF group was significantly higher than that of the other groups (P < 0.05), the 4-cell rate and blastocyst rate in 150 ng/mL bFGF group were significantly lower than those in the other groups (P < 0.05), so 100 ng/mL bFGF was used in subsequent experiments.The 4-cell rate in 3-NPA+bFGF group was significantly higher than that in 3-NPA group (P < 0.05), the blastocyst rate in bFGF group was significantly higher than that of the other groups (P < 0.05).The blastocyst rate in bFGF+3-NPA group was significantly higher than that in 3-NPA group (P < 0.05). Compared with control group, the level of ROS in 3-NPA group was increased significantly (P < 0.05), and the level of ROS in bFGF group was decreased significantly (P < 0.05). GSH and mitochondrial membrane potential were decreased significantly in 3-NPA group (P < 0.05), and were increased significantly in bFGF group (P < 0.05). The level of ROS in bFGF+3-NPA group was significantly lower than that in 3-NPA group (P < 0.05), and the levels of GSH and mitochondrial membrane potential were significantly higher than those in 3-NPA group (P < 0.05).【Conclusion】 The addition of 100 ng/mL bFGF to the in vitro culture medium could reduce the embryonic oxidative stress induced by 3-NPA and improve the function of embryonic mitochondria, so as to improve the in vitro development ability of mouse zygotes.

Key words: mice; embryonic development; basic fibroblast growth factor (bFGF); 3-nitroponic acid (3-NPA); reactive oxygen species

CLC Number:

S813.3

TIAN Wei, FANG Yanya, LIU Haixing, LI Zhongshu. Effect of bFGF on in vitro Development of Mouse Zygotes Treated with 3-nitropropionic Acid[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 608-614.

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Effect of bFGF on in vitro Development of Mouse Zygotes Treated with 3-nitropropionic Acid

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