Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in rats

Abstract Background Premature ovarian failure (POF) is a clinical condition characterized by a diminished ovarian reserve occurring before the age of 40, significantly affecting female reproductive health. However, its exact pathogenesis remains unclear. This research aimed to examine the mechanisms...

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Main Authors: Jiaqi Wu, Yanmeng Wei, Qiangli Peng, Jing Zhu, Huacong Shi, Ting Zhao, Tao Yuan
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Journal of Ovarian Research
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Online Access:https://doi.org/10.1186/s13048-025-01759-3
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author Jiaqi Wu
Yanmeng Wei
Qiangli Peng
Jing Zhu
Huacong Shi
Ting Zhao
Tao Yuan
author_facet Jiaqi Wu
Yanmeng Wei
Qiangli Peng
Jing Zhu
Huacong Shi
Ting Zhao
Tao Yuan
author_sort Jiaqi Wu
collection DOAJ
description Abstract Background Premature ovarian failure (POF) is a clinical condition characterized by a diminished ovarian reserve occurring before the age of 40, significantly affecting female reproductive health. However, its exact pathogenesis remains unclear. This research aimed to examine the mechanisms of cyclophosphamide (CTX)-induced senescence and apoptosis in the ovarian and cerebral cortex tissues of rats to provide insights into delaying aging and protecting female reproductive health. Methods A rat model of POF was established by intraperitoneal injection of CTX. Model efficacy was evaluated by measuring ovarian volume, weight, estrogen, and anti-Müllerian hormone levels. Serum marker changes were detected via enzyme-linked immunosorbent assay (ELISA). Senescence and apoptosis in cerebral cortical and ovarian tissues were observed using β-galactosidase (SA-β-gal) staining and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Western Blot and quantitative polymerase chain reaction (RT-qPCR) were employed to detect the expression levels of cell senescence- and apoptosis-related proteins and genes, verifying the correlation between POF and cellular senescence/apoptosis. Results High-dose CTX induced POF. In rats with POF, the levels of anti-Müllerian hormone (AMH), estradiol (E2), and vitamin D (VD) significantly decreased (P < 0.0001), whereas the levels of testosterone (T) and insulin (INS) significantly increased (P < 0.0001). The number of senescent and apoptotic-positive cells in the ovarian and cerebral cortex tissues of rats with POF was substantially augmented (P < 0.05; P < 0.01). Additionally, the expression of senescence-related proteins cyclin-dependent kinase inhibitor 1 A (CDKN1A), cyclin-dependent kinase inhibitor 2 A (CDKN2A), tumor protein p53 (P53), apoptosis-related protein BCL2-Associated X Protein (Bax), and cysteine-aspartic acid protease 3 (caspase 3) was upregulated. In contrast, the expression of the anti-apoptotic protein BCL-2 was downregulated. The changes ranged from 1.7- to 7.1-fold. These findings demonstrated that high-dose CTX injection leads to cellular senescence and apoptosis, resulting in ovarian pathology. Conclusion High-dose CTX induced POF in rats, resulting in aging and apoptosis in the cerebral cortex and ovarian tissues. Therefore, inhibiting cellular senescence and apoptosis may be a potential approach for restoring ovarian reserve function in POF.
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spelling doaj-art-b5d3ee638fba45a9926eb6b45771ba552025-08-20T03:06:01ZengBMCJournal of Ovarian Research1757-22152025-07-0118111110.1186/s13048-025-01759-3Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in ratsJiaqi Wu0Yanmeng Wei1Qiangli Peng2Jing Zhu3Huacong Shi4Ting Zhao5Tao Yuan6Department of Gynecology, The First People’s Hospital of Yunnan ProvinceDepartment of Gynecology, The First People’s Hospital of Yunnan ProvinceDepartment of Gynecology, Yunnan Provincial Hospital of Traditional Chinese MedicineDepartment of Gynecology, The First People’s Hospital of Yunnan ProvinceDepartment of Gynecology, The First People’s Hospital of Yunnan ProvinceDepartment of Gynecology, The First People’s Hospital of Yunnan ProvinceDepartment of Gynecology, The First People’s Hospital of Yunnan ProvinceAbstract Background Premature ovarian failure (POF) is a clinical condition characterized by a diminished ovarian reserve occurring before the age of 40, significantly affecting female reproductive health. However, its exact pathogenesis remains unclear. This research aimed to examine the mechanisms of cyclophosphamide (CTX)-induced senescence and apoptosis in the ovarian and cerebral cortex tissues of rats to provide insights into delaying aging and protecting female reproductive health. Methods A rat model of POF was established by intraperitoneal injection of CTX. Model efficacy was evaluated by measuring ovarian volume, weight, estrogen, and anti-Müllerian hormone levels. Serum marker changes were detected via enzyme-linked immunosorbent assay (ELISA). Senescence and apoptosis in cerebral cortical and ovarian tissues were observed using β-galactosidase (SA-β-gal) staining and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Western Blot and quantitative polymerase chain reaction (RT-qPCR) were employed to detect the expression levels of cell senescence- and apoptosis-related proteins and genes, verifying the correlation between POF and cellular senescence/apoptosis. Results High-dose CTX induced POF. In rats with POF, the levels of anti-Müllerian hormone (AMH), estradiol (E2), and vitamin D (VD) significantly decreased (P < 0.0001), whereas the levels of testosterone (T) and insulin (INS) significantly increased (P < 0.0001). The number of senescent and apoptotic-positive cells in the ovarian and cerebral cortex tissues of rats with POF was substantially augmented (P < 0.05; P < 0.01). Additionally, the expression of senescence-related proteins cyclin-dependent kinase inhibitor 1 A (CDKN1A), cyclin-dependent kinase inhibitor 2 A (CDKN2A), tumor protein p53 (P53), apoptosis-related protein BCL2-Associated X Protein (Bax), and cysteine-aspartic acid protease 3 (caspase 3) was upregulated. In contrast, the expression of the anti-apoptotic protein BCL-2 was downregulated. The changes ranged from 1.7- to 7.1-fold. These findings demonstrated that high-dose CTX injection leads to cellular senescence and apoptosis, resulting in ovarian pathology. Conclusion High-dose CTX induced POF in rats, resulting in aging and apoptosis in the cerebral cortex and ovarian tissues. Therefore, inhibiting cellular senescence and apoptosis may be a potential approach for restoring ovarian reserve function in POF.https://doi.org/10.1186/s13048-025-01759-3Premature ovarian failureCyclophosphamideApoptosisCellular senescence
spellingShingle Jiaqi Wu
Yanmeng Wei
Qiangli Peng
Jing Zhu
Huacong Shi
Ting Zhao
Tao Yuan
Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in rats
Journal of Ovarian Research
Premature ovarian failure
Cyclophosphamide
Apoptosis
Cellular senescence
title Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in rats
title_full Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in rats
title_fullStr Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in rats
title_full_unstemmed Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in rats
title_short Mechanisms of cell senescence and apoptosis in cyclophosphamide-induced premature ovarian failure in rats
title_sort mechanisms of cell senescence and apoptosis in cyclophosphamide induced premature ovarian failure in rats
topic Premature ovarian failure
Cyclophosphamide
Apoptosis
Cellular senescence
url https://doi.org/10.1186/s13048-025-01759-3
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