Oxidative Stress Injury of Mouse Ovarian Granulosa Cells and Its Effects on Silencing Information Regulator 2-Related Enzyme 1 Signaling Pathway

GONG Haojie; DING Xuemei; BAI Shiping; ZENG Qiufeng; ZHANG Keying; WANG Jianping

doi:10.3969/j.issn.1006-267x.2022.03.053

2022 , Vol. 34 >Issue 3: 1942 - 1954

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.03.053

RESEARCH PAPER

Oxidative Stress Injury of Mouse Ovarian Granulosa Cells and Its Effects on Silencing Information Regulator 2-Related Enzyme 1 Signaling Pathway

GONG Haojie ,
DING Xuemei ,
BAI Shiping ,
ZENG Qiufeng ,
ZHANG Keying ,
WANG Jianping

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Sichuan Key Laboratory of Animal Disease-Resistance Nutrition, Key Laboratory of Animal Disease-Resistance Nutrition and Feed, Ministry of Agriculture and Rural Affairs, Key Laboratory of Animal Disease-Resistance Nutrition, Ministry of Education, Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China

Received date: 2021-08-23

Online published: 2022-03-14

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Abstract

The aim of this study was to explore the effects of different concentrations of hydrogen peroxide (H₂O₂) on oxidative stress injury of mouse ovarian granulosa cells (MOGC), to establish the MOGC oxidative stress model, and to explore the effects of oxidative stress injury on silencing information regulator 2-related enzyme 1 (SIRT1) signaling pathway. A single factor design was adopted and MOGC were treated with 0 (control group), 100, 200, 400 and 800 μmol/L H₂O₂ for 24 h, respectively, and the apoptosis, intracellular reactive oxygen species (ROS) production, related antioxidant enzyme activities, malondialdehyde (MDA) content and apoptosis-related gene expression were determined. The results showed as follows:1) compared with the control group, the cell viability in different concentrations of H₂O₂ groups was significantly decreased (P<0.05), and the cell viability was lower than 50% when H₂O₂ concentration was more than 200 μmol/L and treated for more than 24 h. 2) Compared with the control group, the apoptosis rate in different concentrations of H₂O₂ groups was significantly increased (P<0.05). 3) Compared with the control group, the intracellular ROS production in 200, 400 and 800 μmol/L H₂O₂ groups was significantly increased (P<0.05). 4) Compared with control group, the intracellular superoxide dismutase activity in 400 μmol/L H₂O₂ group was significantly decreased (P<0.05); the intracellular catalase and glutathione peroxidase (GSH-Px) activities and reduced glutathione content in 400 and 800 μmol/L H₂O₂ groups were significantly decreased (P<0.05), while the intracellular MDA content was significantly increased (P<0.05); the intracellular GSH-Px activity in 200 μmol/L H₂O₂ group was significantly decreased (P<0.05), and the intracellular MDA content was significantly increased (P<0.05). 5) Compared with the control group, the intracellular mRNA relative expression levels of B-cell lymphoma-2 interacting mediator of cell death (Bim), cysteine-aspartic protease-3 (Caspase-3) and cysteine-aspartic protease-9 (Caspase-9) in 800 μmol/L H₂O₂ group were significantly increased (P<0.05), while the intracellular mRNA relative expression level of B-cell lymphoma-xL (Bcl-xL) was significantly decreased (P<0.05); the intracellular mRNA relative expression levels of Caspase-3 in 400 μmol/L H₂O₂ group was significantly increased (P<0.05), while the intracellular mRNA relative expression level of Bcl-xL in 400 μmol/L H₂O₂ group was significantly decreased (P<0.05); in addition, the intracellular mRNA relative expression level of SIRT1 in 200, 400 and 800 μmol/L H₂O₂ groups was significantly decreased (P<0.05), while the intracellular mRNA relative expression levels of Forkhead box protein O1 (FoxO1) and p53 were significantly increased (P<0.05). The results suggest that 200 μmol/L H₂O₂ treatment for 24 h is the ideal condition for establishing MOGC oxidative stress model, and SIRT1-FoxO1/p53 pathway is involved in H₂O₂-induced MOGC apoptosis.

Key words： H₂O₂; mouse ovarian granulosa cells; apoptosis; SIRT1

Cite this article

GONG Haojie , DING Xuemei , BAI Shiping , ZENG Qiufeng , ZHANG Keying , WANG Jianping . Oxidative Stress Injury of Mouse Ovarian Granulosa Cells and Its Effects on Silencing Information Regulator 2-Related Enzyme 1 Signaling Pathway[J]. Chinese Journal of Animal Nutrition, 2022 , 34(3) : 1942 -1954 . DOI: 10.3969/j.issn.1006-267x.2022.03.053

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