FLI通过增加糖代谢途径促进牛卵母细胞体外成熟

doi:10.11843/j.issn.0366-6964.2025.06.022

摘要/Abstract

摘要：

为了明确FGF2、LIF和IGF1(FLI)改善牛卵母细胞体外成熟的作用机制。本研究使用屠宰场收集的卵巢，将从卵巢中抽取的卵丘卵母细胞复合体(COCs)随机分为对照组和FLI组，进行体外成熟，每组30枚COCs，每个试验重复3次。体外成熟24 h后检测第一极体排出率、卵丘扩展情况、线粒体膜电位、皮质颗粒分布、培养基中葡萄糖消耗量和丙酮酸含量、卵母细胞氧化还原态、TZPs数量、活性氧含量、相关基因的表达；并统计后续胚胎发育的卵裂率、囊胚率和囊胚细胞数。结果表明，FLI组卵丘扩展指数显著高于对照组(分别为3.16±0.04和2.43±0.02，P < 0.001)。与对照组相比，FLI组卵母细胞的卵丘扩展相关基因表达升高(P < 0.05)，皮质颗粒向卵周迁移增多(P < 0.05)，但是卵母细胞第一极体排出率、线粒体膜电位没有显著变化(P＞0.05)。与对照组相比，FLI组的培养基中葡萄糖的消耗、丙酮酸的含量显著增加(P < 0.05)，卵母细胞内NADPH显著增加(P < 0.001)，并且糖代谢相关基因的表达显著上升(P < 0.05)。添加FLI可以显著降低卵母细胞内氧化还原水平、FAD++和ROS水平(P < 0.05)。IVM 8 h时，FLI组TZPs数量显著高于对照组(分别为106±6.91和78±8.76，P < 0.001)。体外受精后，FLI组卵裂率和囊胚率显著高于对照组(分别为(86.49±0.80)%、(37.44±0.42)%和(74.08±0.91)%、(27.34±1.08)%，P < 0.05)。结果显示，FLI通过增加糖代谢提高了卵母细胞体外成熟质量和胚胎发育。

关键词: 牛, 卵母细胞体外成熟, FLI, 糖代谢

Abstract:

This study aimed to clarify the mechanism of FGF2, LIF and IGF1 (FLI) improving bovine oocyte maturation in vitro. The study used ovaries collected from slaughterhouses, cumulus oocyte complexes (COCs) extracted from ovaries were randomly divided into control group and FLI group for in vitro maturation, 30 COCs per group, each experiment was repeated 3 times. After 24 hours of in vitro maturation, first polar body excretion rate, cumulus expansion, mitochondrial membrane potential, cortical granule distribution, glucose consumption and pyruvate content in the medium, oocyte redox state, number of TZPs, reactive oxygen species content, expression of relevant genes were detected; the cleavage rate, blastocyst rate and blastocyst cell number of subsequent embryo development were counted. The results show that, the cumulus expansion index in FLI group was significantly higher than that in control group (3.16±0.04 and 2.43±0.02, respectively, P < 0.001). Compared with the control group, the expression of cumulus expansion related genes in oocytes of FLI group was increased (P < 0.05), increased migration of cortical granules to periovum (P < 0.05), however, the first polar body excretion rate and mitochondrial membrane potential of oocytes did not change significantly (P>0.05). Compared with the control group, the consumption of glucose and the content of pyruvate in the culture medium of FLI group were significantly increased (P < 0.05), NADPH in oocytes increased significantly (P < 0.001), and the expression of genes related to glucose metabolism showed significant up-regulation (P < 0.05). The addition of FLI could reduce the levels of redox, FAD++ and ROS in oocytes (P < 0.05). At IVM 8 h, the number of TZPs in FLI group was significantly higher than that in control group (106±6.91 and 78±8.76, respectively, P < 0.001). After in vitro fertilization, the cleavage rate and blastocyst rate in FLI group were significantly higher than those in control group ((86.49±0.80)%, (37.44±0.42)% and (74.08±0.91)%, (27.34±1.08)%, respectively, P < 0.05). The results show that, FLI can improve the quality of oocyte in vitro maturation and embryonic development by increasing glucose metabolism.

Key words: bovine, oocyte maturation in vitro, FLI, glycometabolism

中图分类号:

S823.3

韩僖彤, 张楠, 张宁, 张家新. FLI通过增加糖代谢途径促进牛卵母细胞体外成熟[J]. 畜牧兽医学报, 2025, 56(6): 2778-2789.

HAN Xitong, ZHANG Nan, ZHANG Ning, ZHANG Jiaxin. FLI Promotes in Vitro Maturation of Bovine Oocytes by Increasing the Glucose Metabolism Pathway[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2778-2789.

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基因 Gene	引物序列(5′→3′) Primers sequence
PKM	F: CACGCAGAGACCATCAAGAA；R: TCGGATCTCAGGTCCTTTAGT
ACOD1	F: ACTGGAAATGGCTCTCCTCGG；R: CAAGTGGCAGCGTGGATTCTC
MPC1	F: TGAGCTCCGACTAAATGAGGA；R: TGGGATTGACTGCTGGGGGATA
GAPDH	F: GGGTCATCATCTCTGCACCT；R: GGTCATAAGTCCCTCCACGA
LDHA	F: TCTGGATTCAGCTCGCTTCCGTTA；R: TTCTTCAGGGAGACACCAGCAACA
HK1	F: GTGTGCTGTTGATAATCTCC；R: AATAACTGTTGGACGAATGC
GFPT1	F: AAACACAGTCGGCAGTTCCA；R: TGGCTACACCAATCTCAGGC
HAS2	F: CCTCATCATCCAAAGCCTGT；R: CGGGGTAGGTTAGCCTTTTC
TNFAIP6	F: CATCTTGCCACCTACAAGCA；R: CACACCACCACACTCCTTTG
β-actin	F: AACTCCATCATGAAGTGTGACG；R: GATCCACATCTGCTGGAAGG

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