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Autophagy Contributes to Oxidative Stress-Induced Apoptosis in Porcine Granulosa Cells

  • Reproductive Biology: Original Article
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Abstract

Oxidative stress-induced granulosa cell (GC) death is a major cause of follicular atresia. As the major types of programmed cell death, autophagy and apoptosis have been observed in response to H2O2-mediated oxidative stress and have been demonstrated to be responsible for porcine GC death. To date, however, the cellular reactions linking autophagy to the apoptosis of porcine GC under oxidative stress are still poorly understood. Porcine GC were treated with H2O2, and autophagic flux was examined by western blotting. Cell viability and cell death assays were performed after cotreatment of porcine GC with autophagy activator (rapamycin) or inhibitor (3-methyladenine, 3-MA) together with H2O2. We revealed that short exposure (1–3 h) of porcine GC to H2O2 dramatically increased autophagic flux (1.8- to 2.5-fold over that in the control), whereas 6–12 h prolonged treatment decreased autophagy but elevated the caspase-3 activity and GC apoptotic rate. Furthermore, we showed that pretreatment with rapamycin exacerbated H2O2-mediated cytotoxicity and caspase-3 activation but that 3-MA or siRNAs specific for Beclin 1 and Atg7 genes ameliorated H2O2-mediated GC apoptosis. Together, our results indicate that autophagy plays a pivotal role in H2O2-mediated porcine GC apoptosis. Importantly, we show that the early induction of autophagic flux contributes to oxidative stress-induced apoptosis in porcine GC. The results also suggest that regulating the autophagy response in porcine GC under oxidative stress might be a new strategy for abnormal follicular atresia.

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Acknowledgments

The authors would like to thank 2 anonymous reviewers for their constructive comments and suggestions on an earlier version of the manuscript.

Funding

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: this study was supported by the Science and Technology Innovation and Creativity Project of Henan Academy of Agricultural Sciences (NO.2020CX06) and the Independent Innovation Fund Project in Henan Academy of Agricultural Science (NO.2020ZC36).

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Authors and Affiliations

  1. Institute of Animal Husbandry and Veterinary Science, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, China

    Jia-Qing Zhang, Qiao-Ling Ren, Jun-Feng Chen, Bin-Wen Gao, Zi-Jing Zhang, Jing Wang & Bao-Song Xing

  2. Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation, Zhengzhou, 450002, China

    Jia-Qing Zhang, Qiao-Ling Ren, Jun-Feng Chen, Bin-Wen Gao, Zi-Jing Zhang, Jing Wang & Bao-Song Xing

  3. Henan Provincial Animal Husbandry General Station, Zhengzhou, 450008, China

    Xian-Wei Wang & Ze-Jun Xu

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  1. Jia-Qing Zhang
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Contributions

Jia-Qing Zhang and Bao-Song Xing conceived and designed the experiments. Qiao-Ling Ren, Jun-Feng Chen, and Bin-Wen Gao performed the experiments. Xian-Wei Wang and Zi-Jing Zhang analyzed the data. Ze-Jun Xu assisted us to collect samples and supplement the tests. Jia-Qing Zhang and Bao-Song Xing wrote the manuscript.

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Correspondence to Jia-Qing Zhang.

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Animal procedures were conducted in accordance with the guidelines of the Institutional Animal Care and Use Committee of Animal Husbandry and Veterinary Science, Henan Academy of Agricultural Sciences. During the procedure, electrodes were used to ensure that the pigs were in an unconscious state, and then the ovaries were removed.

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Zhang, JQ., Ren, QL., Chen, JF. et al. Autophagy Contributes to Oxidative Stress-Induced Apoptosis in Porcine Granulosa Cells. Reprod. Sci. 28, 2147–2160 (2021). https://doi.org/10.1007/s43032-020-00340-1

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