α-Ketoglutarate Improves Meiotic Maturation of Porcine Oocytes and Promotes the Development of PA Embryos, Potentially by Reducing Oxidative Stress through the Nrf2 Pathway.

Qiang Chen,Leilei Gao,Anmin Lei,Yitian Yuan,Ruibin Wang,Jiannan Li,Yiqi Tian,Joaquin Gadea

doi:10.1155/2022/7113793

Abstract

α-Ketoglutarate (α-KG) is a metabolite in the tricarboxylic acid cycle. It has a strong antioxidant function and can effectively prevent oxidative damage. Previous studies have shown that α-KG exists in porcine follicles, and its content gradually increases as the follicles grow and mature. However, the potential mechanism of supplementation of α-KG on porcine oocytes during in vitro maturation (IVM) has not yet been reported. The purpose of this study was to explore the effect of α-KG on the early embryonic development of pigs and the mechanisms underlying these effects. We found that α-KG can enhance the development of early pig embryos. Adding 20 μM α-KG to the in vitro culture medium significantly increased the rate of blastocyst formation and the total cell number. Compared with to that of the control group, apoptosis in blastocysts of the supplement group was significantly reduced. α-KG reduced the production of reactive oxygen species and glutathione levels in cells. α-KG not only improved the activity of mitochondria but also inhibited the occurrence of apoptosis. After supplementation with α-KG, pig embryo pluripotency-related genes (OCT4, NANOG, and SOX2) and antiapoptotic genes (Bcl2) were upregulated. In terms of mechanism, α-KG activates the Nrf2/ARE signaling pathway to regulate the expression of antioxidant-related targets, thus combating oxidative stress during the in vitro culture of oocytes. Activated Nrf2 promotes the transcription of Bcl2 genes and inhibits cell apoptosis. These results indicate that α-KG supplements have a beneficial effect on IVM by regulating oxidative stress during the IVM of porcine oocytes and can be used as a potential antioxidant for IVM of porcine oocytes.

Highlights

The maturation level of oocytes cultured in vitro and the ability of embryos to develop after fertilization are lower than those of oocytes cultured in vivo, and developmental retardation often occurs
When evaluating the effect of α-KG during the process of pig oocyte maturation, cumulusoocyte complexes (COCs) were cultured with various concentrations of α-ketoglutarate (10, 20, 50, and 100 μM), and the polar body exclusion (PBE) rate of porcine oocytes was evaluated at the end of in vitro maturation (IVM)
We found that supplementation with 20 μM α-ketoglutarate during the IVM process significantly increased the rate of two-cell and blastocyst formation of the PA embryos (Figures 1(a), 1(c), and 1(d); 23:07% ± 1:16% vs. 38:07% ± 1:34% on day 7; P < 0:01) and increased the total cell number (Figure 1(e); 22:67% ± 0:88% vs. 37:67% ± 1:45% on day 7; P < 0:01) of blastocysts compared with that in the control groups

Summary

Introduction

The maturation level of oocytes cultured in vitro and the ability of embryos to develop after fertilization are lower than those of oocytes cultured in vivo, and developmental retardation often occurs. This retardation may occur because oocytes cultured in vitro are sensitive to changes in the external environment, and there are differences between the in vitro culture environment and the internal environment. Porcine embryos produced in vitro have poor developmental potential, resulting in lower blastocyst yield and total cell numbers when compared with their in vivo counterparts [2–4]. We investigated the mechanisms of antioxidative damage and explored the use of effective antioxidants in the culture medium, as an effective way to improve the development of pig embryos in vitro [12]

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