{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T04:05:22Z","timestamp":1787976322988,"version":"build-2784847793"},"reference-count":32,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,4,17]],"date-time":"2023-04-17T00:00:00Z","timestamp":1681689600000},"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. Nutr."],"abstract":"<jats:sec><jats:title>Purpose<\/jats:title><jats:p>Spermatogonial stem cells (SSCs) are essential for maintaining reproductive function in males. <jats:italic>B<\/jats:italic>-lymphoma Mo-MLV insertion region 1 (BMI1) is a vital transcription repressor that regulates cell proliferation and differentiation. However, little is known about the role of BMI1 in mediating the fate of mammalian SSCs and in male reproduction. This study investigated whether BMI1 is essential for male reproduction and the role of alpha-tocopherol (\u03b1-tocopherol), a protective agent for male fertility, as a modulator of BMI1 both <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Methyl thiazolyl tetrazolium (MTT) and 5-ethynyl-2\u2032-deoxyuridine (EDU) assays were used to assess the effect of BMI1 on the proliferative ability of the mouse SSC line C18-4. Real-time polymerase chain reaction (PCR), western blotting, and immunofluorescence were applied to investigate changes in the mRNA and protein expression levels of BMI1. Male mice were used to investigate the effect of \u03b1-tocopherol and a BMI1 inhibitor on reproduction-associated functionality <jats:italic>in vivo<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Analysis revealed that BMI1 was expressed at high levels in testicular tissues and spermatogonia in mice. The silencing of BMI1 inhibited the proliferation of SSCs and DNA synthesis and enhanced the levels of \u03b3-H2AX. \u03b1-tocopherol enhanced the proliferation and DNA synthesis of C18-4 cells, and increased the levels of BMI1. Notably, \u03b1-tocopherol rescued the inhibition of cell proliferation and DNA damage in C18-4 cells caused by the silencing of BMI1. Furthermore, \u03b1-tocopherol restored sperm count (Ctrl vs. PTC-209, <jats:italic>p<\/jats:italic>\u2009=\u20090.0034; Ctrl vs. PTC-209\u2009+\u2009\u03b1-tocopherol, <jats:italic>p<\/jats:italic>\u2009=\u20090.7293) and normalized sperm malformation such as broken heads, irregular heads, lost and curled tails <jats:italic>in vivo<\/jats:italic>, as demonstrated by its antagonism with the BMI1 inhibitor PTC-209.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Analysis demonstrated that \u03b1-tocopherol is a potent <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic> modulator of BMI1, a transcription factor that plays an important role in in SSC proliferation and spermatogenesis. Our findings identify a new target and strategy for treating male infertility that deserves further pre-clinical investigation.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fnut.2023.1141964","type":"journal-article","created":{"date-parts":[[2023,4,17]],"date-time":"2023-04-17T16:36:27Z","timestamp":1681749387000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":7,"title":["Alpha-tocopherol enhances spermatogonial stem cell proliferation and restores mouse spermatogenesis by up-regulating BMI1"],"prefix":"10.3389","volume":"10","author":[{"given":"Mei","family":"Peng","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jintao","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weifan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianlong","family":"Liao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simeng","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Di","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuping","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoping","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,4,17]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1093\/humupd\/dmz006","article-title":"Spermatogonial stem cells: updates from specification to clinical relevance","volume":"25","author":"Sharma","year":"2019","journal-title":"Hum Reprod Update"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1038\/ncb1826","article-title":"Mouse differentiating spermatogonia can generate germinal stem cells in vivo","volume":"11","author":"Barroca","year":"2009","journal-title":"Nat Cell Biol"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1016\/j.omtn.2018.05.015","article-title":"MiR-663a stimulates proliferation and suppresses early apoptosis of human Spermatogonial stem cells by targeting NFIX and regulating cell cycle","volume":"12","author":"Zhou","year":"2018","journal-title":"Mol Ther Nucleic Acids"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1016\/j.stemcr.2014.08.004","article-title":"2014 generation of haploid spermatids with fertilization and development capacity from human spermatogonial stem cells of cryptorchid patients","volume":"3","author":"Yang","year":"2014","journal-title":"Stem Cell Rep"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1016\/j.semcdb.2014.04.006","article-title":"Regulation of the germ stem cell niche as the foundation for adult spermatogenesis: a role for miRNAs?","volume":"29","author":"van den Driesche","year":"2014","journal-title":"Semin Cell Dev Biol"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"970","DOI":"10.1093\/humupd\/dmab025","article-title":"Reply: Spermatogonia stem cell technology: a new avenue for all age Klinefelter patients","volume":"27","author":"Deebel","year":"2021","journal-title":"Hum Reprod Update"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1833","DOI":"10.1172\/JCI65822","article-title":"Eliminating malignant contamination from therapeutic human spermatogonial stem cells","volume":"123","author":"Dovey","year":"2013","journal-title":"J Clin Invest"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"103193","DOI":"10.1016\/j.ebiom.2020.103193","article-title":"BMI1 in the heart: novel functions beyond tumorigenesis","volume":"63","author":"Yang","year":"2021","journal-title":"EBioMedicine"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1172\/jci20800","article-title":"Bmi1, stem cells, and senescence regulation","volume":"113","author":"Park","year":"2004","journal-title":"J Clin Invest"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1172\/JCI92862","article-title":"2017 Polycomb subunit BMI1 determines uterine progesterone responsiveness essential for normal embryo implantation","volume":"128","author":"Xin","year":"2018","journal-title":"J Clin Invest"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"1200","DOI":"10.1002\/stem.3007","article-title":"Bmi1 overexpression in mesenchymal stem cells exerts antiaging and Antiosteoporosis effects by inactivating p16\/p19 signaling and inhibiting oxidative stress","volume":"37","author":"Chen","year":"2019","journal-title":"Stem Cells"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"583","DOI":"10.1002\/jbmr.3921","article-title":"2019 the Polycomb protein Bmi1 plays a crucial role in the prevention of 1,25 (OH)2 D deficiency-induced bone loss","volume":"35","author":"Sun","year":"2020","journal-title":"J Bone Miner Res"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1002\/path.5244","article-title":"Bmi1 marks gastric stem cells located in the isthmus in mice","volume":"248","author":"Yoshioka","year":"2019","journal-title":"J Pathol"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"358","DOI":"10.7150\/ijbs.23325","article-title":"Bmi1 deficient mice exhibit male infertility","volume":"14","author":"Dai","year":"2018","journal-title":"Int J Biol Sci"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1016\/j.bbrc.2021.10.074","article-title":"BMI1 promotes spermatogonia proliferation through epigenetic repression of Ptprm","volume":"583","author":"Zhang","year":"2021","journal-title":"Biochem Biophys Res Commun"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"650","DOI":"10.1126\/science.56.1458.650","article-title":"On the existence of a hitherto unrecognized dietary factor essential for reproduction","volume":"56","author":"Evans","year":"1922","journal-title":"Science"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"794","DOI":"10.1096\/fj.05-5385fje","article-title":"Phospholipid transfer protein deficiency reduces sperm motility and impairs fertility of mouse males","volume":"20","author":"Drouineaud","year":"2006","journal-title":"FASEB J"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"1149","DOI":"10.1016\/j.fertnstert.2010.10.005","article-title":"Vitamin E supplementation in semen-freezing medium improves the motility and protects sperm from freeze-thaw-induced DNA damage","volume":"95","author":"Kalthur","year":"2011","journal-title":"Fertil Steril"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1093\/ajcn\/nqaa124","article-title":"Effects of coadministration of DHA and vitamin E on spermatogram, seminal oxidative stress, and sperm phospholipids in asthenozoospermic men: a randomized controlled trial","volume":"112","author":"Eslamian","year":"2020","journal-title":"Am J Clin Nutr"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"1780","DOI":"10.3390\/antiox10111780","article-title":"Vitamin E delivery systems increase resistance to oxidative stress in Red Deer sperm cells: hydrogel and Nanoemulsion carriers","volume":"10","author":"Jurado-Campos","year":"2021","journal-title":"Antioxidants (Basel)"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.freeradbiomed.2020.07.024","article-title":"Nanotechnology in reproduction: vitamin E nanoemulsions for reducing oxidative stress in sperm cells","volume":"160","author":"S\u00e1nchez-Rubio","year":"2020","journal-title":"Free Radic Biol Med"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"101289","DOI":"10.1016\/j.psj.2021.101289","article-title":"Dietary supplementation of pumpkin seed oil and sunflower oil along with vitamin E improves sperm characteristics and reproductive hormones in roosters","volume":"100","author":"Lotfi","year":"2021","journal-title":"Poult Sci"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"101635","DOI":"10.1016\/j.psj.2021.101635","article-title":"Enhancement of sperm quality and fertility-related parameters in Hubbard grandparent rooster fed diets supplemented with soybean lecithin and vitamin E","volume":"101","author":"Shabani","year":"2022","journal-title":"Poult Sci"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"9910","DOI":"10.3390\/ijms16059910","article-title":"Effects of dietary vitamin E on fertility functions in poultry species","volume":"16","author":"Rengaraj","year":"2015","journal-title":"Int J Mol Sci"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"2580","DOI":"10.1002\/stem.198","article-title":"Nodal signaling via an autocrine pathway promotes proliferation of mouse spermatogonial stem\/progenitor cells through Smad2\/3 and Oct-4 activation","volume":"27","author":"He","year":"2009","journal-title":"Stem Cells"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.mce.2008.01.008","article-title":"Expression localization of Bmi1 in mice testis","volume":"287","author":"Zhang","year":"2008","journal-title":"Mol Cell Endocrinol"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"684","DOI":"10.1038\/modpathol.3800577","article-title":"Variability in the expression of polycomb proteins in different normal and tumoral tissues. A pilot study using tissue microarrays","volume":"19","author":"S\u00e1nchez-Beato","year":"2006","journal-title":"Mod Pathol"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"302","DOI":"10.1038\/nature01587","article-title":"Bmi-1 is required for maintenance of adult self-renewing haematopoietic stem cells","volume":"423","author":"Park","year":"2003","journal-title":"Nature"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1016\/S0304-3835(98)00192-X","article-title":"Cloning of the rat proto-oncogene bmi-1","volume":"133","author":"Osaka","year":"1998","journal-title":"Cancer Lett"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"270","DOI":"10.1038\/nchembio730","article-title":"A carbohydrate-antioxidant hybrid polymer reduces oxidative damage in spermatozoa and enhances fertility","volume":"1","author":"Fleming","year":"2005","journal-title":"Nat Chem Biol"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"1151","DOI":"10.1016\/j.fertnstert.2013.06.019","article-title":"Antioxidants enhance the recovery of three cycles of bleomycin, etoposide, and cisplatin-induced testicular dysfunction, pituitary-testicular axis, and fertility in rats","volume":"100","author":"Kilarkaje","year":"2013","journal-title":"Fertil Steril"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1359\/jbmr.090812","article-title":"Defects in mesenchymal stem cell self-renewal and cell fate determination lead to an osteopenic phenotype in Bmi-1 null mice","volume":"25","author":"Zhang","year":"2010","journal-title":"J Bone Miner Res"}],"container-title":["Frontiers in Nutrition"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnut.2023.1141964\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,4,17]],"date-time":"2023-04-17T16:36:30Z","timestamp":1681749390000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fnut.2023.1141964\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,17]]},"references-count":32,"alternative-id":["10.3389\/fnut.2023.1141964"],"URL":"https:\/\/doi.org\/10.3389\/fnut.2023.1141964","relation":{},"ISSN":["2296-861X"],"issn-type":[{"value":"2296-861X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,4,17]]},"article-number":"1141964"}}