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PS48 通过激活 PI3K/Akt 信号通路促进猪卵母细胞的体外成熟和发育能力。

PS48 promotes in vitro maturation and developmental competence of porcine oocytes through activating PI3K/Akt signalling pathway.

机构信息

State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China.

School of Animal Science and Technology, Guangxi University, Nanning, China.

出版信息

Reprod Domest Anim. 2020 Dec;55(12):1678-1687. doi: 10.1111/rda.13818. Epub 2020 Nov 10.

Abstract

Oocyte maturation plays a vitally important role in porcine reproduction. Regrettably, the quality of oocytes matured in vitro is weaker than that of in vivo matured oocytes. We collected and cultivated porcine cumulus oocyte complexes (COCs) in vitro with phosphoinositide-dependent kinase 1 (PDK1) activator 5-(4-chloro-phenyl)-3-phenyl-pent-2-enoic acid (PS48), whose concentrations were 0, 2, 5, 10 and 20 µM to investigate whether the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) signalling pathway would impact the oocyte quality. The results showed that 10 µM PS48 increased the oocyte proportion of metaphase II (MII) stage and improved the expansion of cumulus cells (CCs). What's more, the activation of PI3K/Akt signalling pathway could regulate the expression of maturation-related genes and proteins. The results of quantitative real-time PCR showed that 10 µM PS48 increased the mRNA and protein levels of Akt and regulated maturation-related genes, including cyclin B1, MOS, BMP15, GDF9, CDC2, mTOR, BAX, BCL2 and caspase-3. The results of Western blot indicated that 10µM PS48 increased the protein abundance of Akt, phosphorylation of Akt Thr308 (p-Akt ) and cyclin B1, but decreased the protein abundance of pro-apoptotic BAX. These results suggested that adding 10 µM PS48 to mature culture medium could promote the maturation of porcine oocytes, potentially through activating the PI3K/Akt signalling pathway.

摘要

卵母细胞成熟在猪繁殖中起着至关重要的作用。遗憾的是,体外成熟的卵母细胞的质量比体内成熟的卵母细胞差。我们收集并培养了猪卵丘-卵母细胞复合物(COC),用磷脂酰肌醇-3-激酶/蛋白激酶 B(PI3K/Akt)信号通路的激活剂 5-(4-氯苯基)-3-苯基戊-2-烯酸(PS48),浓度分别为 0、2、5、10 和 20μM,研究其是否会影响卵母细胞的质量。结果表明,10μM PS48 增加了中期 II(MII)期卵母细胞的比例,并改善了卵丘细胞(CC)的扩展。此外,PI3K/Akt 信号通路的激活可以调节成熟相关基因和蛋白质的表达。实时定量 PCR 结果表明,10μM PS48 增加了 Akt 的 mRNA 和蛋白水平,并调节了成熟相关基因,包括 cyclin B1、MOS、BMP15、GDF9、CDC2、mTOR、BAX、BCL2 和 caspase-3。Western blot 结果表明,10μM PS48 增加了 Akt 的蛋白丰度、Akt Thr308 磷酸化(p-Akt)和 cyclin B1 的蛋白丰度,但降低了促凋亡 BAX 的蛋白丰度。这些结果表明,在成熟培养基中添加 10μM PS48 可以促进猪卵母细胞的成熟,可能是通过激活 PI3K/Akt 信号通路。

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