Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation
Background/Aims: This study investigated the effect of consecutive superovulation on the ovaries and established a premature ovarian failure (POF) model in mice. Methods: The mouse POF model was induced by 5-15 consecutive superovulation treatments with pregnant mare serum gonadotropin (PMSG), human...
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Cell Physiol Biochem Press GmbH & Co KG
2018-12-01
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doaj-c5d537c5c14340fda913702ad90b9a332020-11-24T21:52:57ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782018-12-015152341235810.1159/000495895495895Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive SuperovulationXiaowei NieYoujin DaiYuan ZhengDan BaoQin ChenYuan YinHeling FuDaorong HouBackground/Aims: This study investigated the effect of consecutive superovulation on the ovaries and established a premature ovarian failure (POF) model in mice. Methods: The mouse POF model was induced by 5-15 consecutive superovulation treatments with pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG) and prostaglandin F2α (PGF2α). Normal adult mice were compared with mice displaying natural ovarian aging. The following serum biochemical parameters were measured: including follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P), estradiol (E2), inhibin B (INH B), malondialdehyde (MDA), total superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels. Follicles were counted using H&E staining. Levels of 8-hydroxyguanosine (8-OhdG), 4-hydroxynonenal (4-HNE), nitrotyrosine (NTY), anti-Mullerian hormone (AMH) and CDKN2A/ p16 (p16) were detected using immunohistochemical staining. Reactive oxygen species (ROS) levels were measured using dihydroethidium (DHE) staining. Cell apoptosis was detected using an in situ TUNEL fluorescence staining assay. Levels of proteins involved in ROS-related pathways and the p16 protein were detected using Western blotting. Sod1, Sod2 and Sod3 mRNA levels were detected using quantitative polymerase chain reaction (Q-PCR). Oocyte quality was evaluated using in vitro fertilization (IVF) and zygote culture. Results: Consecutive superovulation groups presented lower P, E2, SOD, GSH-Px and INH B levels, significantly higher FSH, LH, MDA and ROS levels, and significantly fewer primordial follicles compared with the control group. Consecutive superovulation groups presented significantly increased levels of Sod2, 8-OhdG, 4-HNE, NTY, significantly increased levels of the SIRT1 and FOXO1 proteins, significantly increased levels of the senescence-associated protein p16, as well as decreased AMH, Sod1 and Sod3 levels and increased granulosa cell apoptosis compared with the control group. Conclusion: Consecutive superovulation significantly decreased ovarian function and oocyte quality and increased oxidative stress and apoptosis in the ovary via a mechanism involving the p16 and SIRT1/FOXO1 signaling pathways. These findings suggest that consecutive superovulation may be used to establish a mouse model of ovarian aging.https://www.karger.com/Article/FullText/495895OvulationOxidative stressApoptosisPremature ovarian failureOvary aging |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Xiaowei Nie Youjin Dai Yuan Zheng Dan Bao Qin Chen Yuan Yin Heling Fu Daorong Hou |
spellingShingle |
Xiaowei Nie Youjin Dai Yuan Zheng Dan Bao Qin Chen Yuan Yin Heling Fu Daorong Hou Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation Cellular Physiology and Biochemistry Ovulation Oxidative stress Apoptosis Premature ovarian failure Ovary aging |
author_facet |
Xiaowei Nie Youjin Dai Yuan Zheng Dan Bao Qin Chen Yuan Yin Heling Fu Daorong Hou |
author_sort |
Xiaowei Nie |
title |
Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation |
title_short |
Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation |
title_full |
Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation |
title_fullStr |
Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation |
title_full_unstemmed |
Establishment of a Mouse Model of Premature Ovarian Failure Using Consecutive Superovulation |
title_sort |
establishment of a mouse model of premature ovarian failure using consecutive superovulation |
publisher |
Cell Physiol Biochem Press GmbH & Co KG |
series |
Cellular Physiology and Biochemistry |
issn |
1015-8987 1421-9778 |
publishDate |
2018-12-01 |
description |
Background/Aims: This study investigated the effect of consecutive superovulation on the ovaries and established a premature ovarian failure (POF) model in mice. Methods: The mouse POF model was induced by 5-15 consecutive superovulation treatments with pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG) and prostaglandin F2α (PGF2α). Normal adult mice were compared with mice displaying natural ovarian aging. The following serum biochemical parameters were measured: including follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P), estradiol (E2), inhibin B (INH B), malondialdehyde (MDA), total superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels. Follicles were counted using H&E staining. Levels of 8-hydroxyguanosine (8-OhdG), 4-hydroxynonenal (4-HNE), nitrotyrosine (NTY), anti-Mullerian hormone (AMH) and CDKN2A/ p16 (p16) were detected using immunohistochemical staining. Reactive oxygen species (ROS) levels were measured using dihydroethidium (DHE) staining. Cell apoptosis was detected using an in situ TUNEL fluorescence staining assay. Levels of proteins involved in ROS-related pathways and the p16 protein were detected using Western blotting. Sod1, Sod2 and Sod3 mRNA levels were detected using quantitative polymerase chain reaction (Q-PCR). Oocyte quality was evaluated using in vitro fertilization (IVF) and zygote culture. Results: Consecutive superovulation groups presented lower P, E2, SOD, GSH-Px and INH B levels, significantly higher FSH, LH, MDA and ROS levels, and significantly fewer primordial follicles compared with the control group. Consecutive superovulation groups presented significantly increased levels of Sod2, 8-OhdG, 4-HNE, NTY, significantly increased levels of the SIRT1 and FOXO1 proteins, significantly increased levels of the senescence-associated protein p16, as well as decreased AMH, Sod1 and Sod3 levels and increased granulosa cell apoptosis compared with the control group. Conclusion: Consecutive superovulation significantly decreased ovarian function and oocyte quality and increased oxidative stress and apoptosis in the ovary via a mechanism involving the p16 and SIRT1/FOXO1 signaling pathways. These findings suggest that consecutive superovulation may be used to establish a mouse model of ovarian aging. |
topic |
Ovulation Oxidative stress Apoptosis Premature ovarian failure Ovary aging |
url |
https://www.karger.com/Article/FullText/495895 |
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