{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T00:23:22Z","timestamp":1776731002692,"version":"3.51.2"},"reference-count":53,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,7,16]],"date-time":"2025-07-16T00:00:00Z","timestamp":1752624000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Recent studies have focused on investigating the role of cellular senescence in ovarian aging. Targeting cellular senescence has been proposed as a potential strategy to improve ovarian aging. p16 and p21 are classical molecules involved in mediating cellular senescence. In our previous study, we demonstrated that ablation of p16 is dispensable for premature ovarian aging induced by alkylating agents. In the present study, we investigated whether p21 deficiency could mitigate ovarian aging caused by alkylating agents.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Eight-week-old wild-type (WT, n=7) and p21 knockout (KO, n=7) female mice received a single injection of busulfan (BUL, 30 mg\/kg) and cyclophosphamide (CTX, 120 mg\/kg) to induce premature ovarian insufficiency (POI). Untreated WT (n=4) and p21 KO (n=4) mice served as controls. Ovaries were analyzed thirteen weeks after treatment. Ovarian reserve, folliculogenesis, cell proliferation, apoptosis and senescence, multinucleated giant cells (MGCs) and their characteristics, pro-inflammatory factors, fibrosis, ovarian stromal cell properties, and the expression of cell cycle inhibitors, including p16, p19, p27, and p53, were evaluated.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Female mice treated with alkylating agents exhibited typical features of POI, including a dramatic reduction in the number of primordial and growing follicles; defective folliculogenesis characterized by growth arrest in early-stage follicles, extensive atresia in mid-stage follicles, dysregulated FSH receptor (FSHr) expression in antral follicles, and abnormal over-activation of primordial follicles; the presence of hemosiderin-laden MGCs and fibrosis in the ovarian cortical region. p21 deficiency did not significantly mitigate these phenotypes. There were no significantly differences in the expression of pro-inflammatory factors, folliculogenesis-regulating factors, or steroidogenesis-related factors and cell cycle inhibitors between WT and p21 KO mice treated with alkylating agents. In\u00a0addition, p21 deficiency did not prevent alkylating agent-induced cellular senescence.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>These results demonstrated that p21 is dispensable for POI caused by alkylating agents, suggesting that targeting p21 alone may not mitigate ovarian aging caused by alkylating agents.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2025.1616965","type":"journal-article","created":{"date-parts":[[2025,7,16]],"date-time":"2025-07-16T04:12:23Z","timestamp":1752639143000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Loss of p21 does not protect against premature ovarian insufficiency caused by alkylating agents"],"prefix":"10.3389","volume":"16","author":[{"given":"Xiaohui","family":"Lu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongli","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiaming","family":"Song","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qin","family":"Wan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pengfei","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yufeng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pingping","family":"Xue","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiuliang","family":"Dai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,7,16]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1093\/humupd\/dmz027","article-title":"Ovarian damage from chemotherapy and current approaches to its protection","volume":"25","author":"Spears","year":"2019","journal-title":"Hum Reprod Update"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1177\/1933719119831784","article-title":"Diminished ovarian reserve chemotherapy-induced mouse model: A tool for the preclinical assessment of new therapies for ovarian damage","volume":"27","author":"Buigues","year":"2019","journal-title":"Reprod Sci"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1016\/j.yrtph.2016.09.002","article-title":"The comparison of animal models for premature ovarian failure established by several different source of inducers","volume":"81","author":"Zhang","year":"2016","journal-title":"Regul Toxicol Pharmacol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1002\/ame2.12477","article-title":"An overview of different methods to establish a murine premature ovarian failure model","volume":"7","author":"Pouladvand","year":"2024","journal-title":"Anim Model Exp Med"},{"key":"B5","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1186\/s12884-023-05476-x","article-title":"Loss of P16 does not protect against premature ovarian insufficiency caused by alkylating agents","volume":"23","author":"Liu","year":"2023","journal-title":"BMC Pregnancy Childbirth"},{"key":"B6","doi-asserted-by":"publisher","first-page":"23178","DOI":"10.1038\/s41598-024-73828-4","article-title":"The senolytic drug Abt-263 accelerates ovarian aging in older female mice","volume":"14","author":"Xia","year":"2024","journal-title":"Sci Rep"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1517\/13543776.17.9.1061","article-title":"Alkylating agents and cancer therapy","volume":"17","author":"Ralhan","year":"2007","journal-title":"Expert Opin Ther Patents"},{"key":"B8","doi-asserted-by":"publisher","first-page":"185ra62","DOI":"10.1126\/scitranslmed.3005402","article-title":"Cyclophosphamide triggers follicle activation and \u201cBurnout\u201d; As101 prevents follicle loss and preserves fertility","volume":"5","author":"Kalich-Philosoph","year":"2013","journal-title":"Sci Transl Med"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2020.11.040","article-title":"Ovaries of patients recently treated with alkylating agent chemotherapy indicate the presence of acute follicle activation, elucidating its role among other proposed mechanisms of follicle loss","volume":"115","author":"Shai","year":"2021","journal-title":"Fertil Steril"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1530\/JOE-18-0370","article-title":"Inhibitors of apoptosis protect the ovarian reserve from cyclophosphamide","volume":"240","author":"Luan","year":"2019","journal-title":"J Endocrinol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1631\/jzus.B1200181","article-title":"Accelerated ovarian aging in mice by treatment of busulfan and cyclophosphamide","volume":"14","author":"Jiang","year":"2013","journal-title":"J Zhejiang Univ Sci B"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2022.781404","article-title":"Pqq dietary supplementation prevents alkylating agent-induced ovarian dysfunction in mice","volume":"13","author":"Dai","year":"2022","journal-title":"Front Endocrinol (Lausanne)"},{"key":"B13","doi-asserted-by":"publisher","first-page":"590","DOI":"10.1515\/mr-2022-0031","article-title":"Ovarian aging: mechanisms and intervention strategies","volume":"2","author":"Zhu","year":"2022","journal-title":"Med Rev (2021)"},{"key":"B14","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1038\/s41514-024-00157-1","article-title":"The role of cellular senescence in ovarian aging","volume":"10","author":"Hense","year":"2024","journal-title":"NPJ Aging"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1262\/jrd.2023-021","article-title":"Accumulation of senescent cells in the stroma of aged mouse ovary","volume":"69","author":"Maruyama","year":"2023","journal-title":"J Reprod Dev"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/j.mad.2020.111425","article-title":"Cellular hallmarks of aging emerge in the ovary prior to primordial follicle depletion","volume":"194","author":"Ansere","year":"2021","journal-title":"Mech Ageing Dev"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1155\/2022\/9144644","article-title":"Senotherapy protects against cisplatin-induced ovarian injury by removing senescent cells and alleviating DNA damage","volume":"2022","author":"Du","year":"2022","journal-title":"Oxid Med Cell Longev"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1172\/JCI95148","article-title":"Mechanisms and functions of cellular senescence","volume":"128","author":"Herranz","year":"2018","journal-title":"J Clin Invest"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.3390\/ijms242417193","article-title":"The role of cellular senescence in cyclophosphamide-induced primary ovarian insufficiency","volume":"24","author":"Xu","year":"2023","journal-title":"Int J Mol Sci"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1038\/s43587-023-00532-9","article-title":"Nadase Cd38 is a key determinant of ovarian aging","volume":"4","author":"Yang","year":"2024","journal-title":"Nat Aging"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1038\/s43587-024-00607-1","article-title":"Spatiotemporal transcriptomic changes of human ovarian aging and the regulatory role of Foxp1","volume":"4","author":"Wu","year":"2024","journal-title":"Nat Aging"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2017.231","article-title":"Oocyte-specific deletion of furin leads to female infertility by causing early secondary follicle arrest in mice","volume":"8","author":"Meng","year":"2017","journal-title":"Cell Death Dis"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1177\/37.3.2918221","article-title":"Alpha-smooth muscle actin, a differentiation marker of smooth muscle cells, is present in microfilamentous bundles of pericytes","volume":"37","author":"Skalli","year":"1989","journal-title":"J Histochem Cytochem"},{"key":"B24","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1177\/0192623315613452","article-title":"Pdgfr inhibition results in pericyte depletion and hemorrhage into the corpus luteum of the rat ovary","volume":"44","author":"Hall","year":"2016","journal-title":"Toxicol Pathol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1530\/REP-20-0489","article-title":"Macrophage-derived multinucleated giant cells: hallmarks of the aging ovary","volume":"161","author":"Foley","year":"2021","journal-title":"Reproduction"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1530\/REP-23-0499","article-title":"Reproductive ageing: inflammation, immune cells, and cellular senescence in the aging ovary","volume":"168","author":"Isola","year":"2024","journal-title":"Reproduction"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/deae132","article-title":"Unveiling the role of chronic inflammation in ovarian aging: insights into mechanisms and clinical implications","volume":"39","author":"Zeng","year":"2024","journal-title":"Hum Reprod"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1530\/REP-16-0129","article-title":"Reproductive age-associated fibrosis in the stroma of the mammalian ovary","volume":"152","author":"Briley","year":"2016","journal-title":"Reproduction"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-319-31973-5_7","article-title":"Ovarian folliculogenesis","volume":"58","author":"Rimon-Dahari","year":"2016","journal-title":"Results Probl Cell Differ"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1210\/me.2007-0172","article-title":"P27kip1 (Cyclin-dependent kinase inhibitor 1b) controls ovarian development by suppressing follicle endowment and activation and promoting follicle atresia in mice","volume":"21","author":"Rajareddy","year":"2007","journal-title":"Mol Endocrinol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1262\/jrd.10-119h","article-title":"P27(Kip1) negatively regulates the activation of murine primordial oocytes","volume":"57","author":"Hirashima","year":"2011","journal-title":"J Reprod Dev"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M301206200","article-title":"The cyclin-dependent kinase inhibitors P27kip1 and P21cip1 cooperate to restrict proliferative life span in differentiating ovarian cells","volume":"278","author":"Jirawatnotai","year":"2003","journal-title":"J Biol Chem"},{"key":"B33","doi-asserted-by":"publisher","first-page":"19750","DOI":"10.1038\/s41598-020-76543-y","article-title":"Amh has no role in predicting oocyte quality in women with advanced age undergoing Ivf\/Icsi cycles","volume":"10","author":"Dai","year":"2020","journal-title":"Sci Rep"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.3109\/09513599909167588","article-title":"Regulation of granulosa cell proliferation and apoptosis during follicular development","volume":"13","author":"Maruo","year":"1999","journal-title":"Gynecol Endocrinol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1007\/BF02899632","article-title":"Granulosa cell proliferation differentiation and its role in follicular development","volume":"50","author":"Cuiling","year":"2005","journal-title":"Chin Sci Bull"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1089\/scd.2021.0005","article-title":"Matrigel\/umbilical cord-derived mesenchymal stem cells promote granulosa cell proliferation and ovarian vascularization in a mouse model of premature ovarian failure","volume":"30","author":"Zhou","year":"2021","journal-title":"Stem Cells Dev"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2024.112248","article-title":"Fgf2 promotes the proliferation of injured granulosa cells in premature ovarian failure via hippo-yap signaling pathway","volume":"589","author":"Cheng","year":"2024","journal-title":"Mol Cell Endocrinol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.7150\/thno.89878","article-title":"Polycomb repressive complex 1 modulates granulosa cell proliferation in early folliculogenesis to support female reproduction","volume":"14","author":"Gao","year":"2024","journal-title":"Theranostics"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/deae297","article-title":"Elevated plasma molybdenum level increases the risk of premature ovarian insufficiency through vascular endothelial injury","volume":"40","author":"Wang","year":"2025","journal-title":"Hum Reprod"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1016\/j.animal.2023.100743","article-title":"Review: roles of follicle-stimulating hormone in preantral folliculogenesis of domestic animals: what can we learn from model species and where do we go from here","author":"Morton","year":"2023","journal-title":"Animal"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1183\/09031936.00119108","article-title":"Haemosiderin-laden macrophages in the bronchoalveolar lavage fluid of patients with diffuse alveolar damage","volume":"33","author":"Maldonado","year":"2009","journal-title":"Eur Respir J"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/dem027","article-title":"Cortical fibrosis and blood-vessels damage in human ovaries exposed to chemotherapy. Potential mechanisms of ovarian injury","volume":"22","author":"Meirow","year":"2007","journal-title":"Hum Reprod"},{"key":"B43","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1038\/s41467-024-55509-y","article-title":"Physiological premature aging of ovarian blood vessels leads to decline in fertility in middle-aged mice","volume":"16","author":"Mu","year":"2025","journal-title":"Nat Commun"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2567.2000.00025.x","article-title":"Inflammatory cytokine production by immunological and foreign body multinucleated giant cells","volume":"100","author":"Hernandez-Pando","year":"2000","journal-title":"Immunology"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abn4564","article-title":"Female reproductive life span is extended by targeted removal of fibrotic collagen from the mouse ovary","volume":"8","author":"Umehara","year":"2022","journal-title":"Sci Adv"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1016\/j.mocell.2024.100113","article-title":"The role of P21 in cellular senescence and aging-related diseases","volume":"47","author":"Yan","year":"2024","journal-title":"Mol Cells"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1158\/1541-7786.MCR-08-0373","article-title":"P53 deficiency leads to compensatory up-regulation of P16ink4a","volume":"7","author":"Leong","year":"2009","journal-title":"Mol Cancer Res"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1155\/2021\/6634718","article-title":"Advanced oxidation protein products induce G1\/G0-phase arrest in ovarian granulosa cells via the Ros-Jnk\/P38 Mapk-P21 pathway in premature ovarian insufficiency","volume":"2021","author":"Zhou","year":"2021","journal-title":"Oxid Med Cell Longev"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.3892\/etm.2015.2498","article-title":"Tripterygium glycosides induce premature ovarian failure in rats by promoting P53 phosphorylation and activating the serine\/threonine kinase 11-P53-P21 signaling pathway","volume":"10","author":"Liu","year":"2015","journal-title":"Exp Ther Med"},{"key":"B50","doi-asserted-by":"publisher","first-page":"308","DOI":"10.3390\/antiox13030308","article-title":"Trim28-mediated excessive oxidative stress induces cellular senescence in granulosa cells and contributes to premature ovarian insufficiency in vitro and in vivo","volume":"13","author":"Zhou","year":"2024","journal-title":"Antioxidants (Basel)"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1007\/s11357-023-00728-2","article-title":"Increased cellular senescence in doxorubicin-induced murine ovarian injury: effect of senolytics","volume":"45","author":"Gao","year":"2023","journal-title":"Geroscience"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1038\/s43587-024-00762-5","article-title":"Molecular and genetic insights into human ovarian aging from single-nuclei multi-omics analyses","volume":"5","author":"Jin","year":"2025","journal-title":"Nat Aging"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1007\/s11357-022-00573-9","article-title":"Senolytic treatment reverses obesity-mediated senescent cell accumulation in the ovary","volume":"44","author":"Hense","year":"2022","journal-title":"Geroscience"}],"container-title":["Frontiers in Endocrinology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2025.1616965\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,16]],"date-time":"2025-07-16T04:12:24Z","timestamp":1752639144000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2025.1616965\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,16]]},"references-count":53,"alternative-id":["10.3389\/fendo.2025.1616965"],"URL":"https:\/\/doi.org\/10.3389\/fendo.2025.1616965","relation":{},"ISSN":["1664-2392"],"issn-type":[{"value":"1664-2392","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,16]]},"article-number":"1616965"}}