{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T22:55:28Z","timestamp":1790031328840,"version":"4.0.1"},"reference-count":68,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2022,11,28]],"date-time":"2022-11-28T00:00:00Z","timestamp":1669593600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81971442"],"award-info":[{"award-number":["81971442"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Andrology"],"published-print":{"date-parts":[[2023,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Backgrounds<\/jats:title><jats:p>The Sertoli cell that plays a vital role during spermatogenesis is a known target of physiological and pathological factors affecting testicular development. Tumor necrosis factor alpha (TNF\u03b1) participates in the blood\u2013testis barrier reconstruction, cell apoptosis, and inflammatory response by recognizing receptors on Sertoli cell. TNF\u03b1 has also been shown to induce the proliferation of immature Sertoli cell in vitro, yet the mechanism still remains unclarified.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>This study was designed to investigate the effect of TNF\u03b1 on blood\u2013testis barrier development during puberty and the underlying mechanisms of TNF\u03b1\u2010induced immature Sertoli cell proliferation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and methods<\/jats:title><jats:p>Immature male Sprague\u2013Dawley rats of postnatal day 12 were intraperitoneally injected with TNF\u03b1. Biotin\u2010labeled method was used to detect permeability of the developing blood\u2013testis barrier after TNF\u03b1 treatment, and the distribution of occludin and junctional adhesion molecule\u2010A (JAM\u2010A) were detected by immunofluorescence. Sertoli cells isolated from Sprague\u2013Dawley rats of postnatal day 10 were cultured in vitro and treated with TNF\u03b1. Cell proliferation rate was reflected by Cell Counting Kit\u20108 (CCK\u20108) and 5\u2010ethynyl\u20102'\u2010deoxyuridine (EdU) assay. Immunoblot and quantitative polymerase chain reaction were used to detect the expression of proliferating cell nuclear antigen, Fbxo4, and cyclin D1. Immunoprecipitation was used to detect the ubiquitination of cyclin D1 and the interaction between Fbxo4 and cyclin D1. Ammonium pyrrolidinedithiocarbamate (PDTC) was applied to detect the effect of nuclear factor kappaB (NF\u03baB) activity inhibition on TNF\u03b1\u2010induced Sertoli cell proliferation. The adenoviral recombinant plasmid containing rat Fbxo4 gene was constructed to investigate the effect of Fbxo4 overexpression on Sertoli cell proliferation promoted by TNF\u03b1.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The in vivo experiment revealed a significant delay of blood\u2013testis barrier maturation in pubertal rats caused by exogenous TNF\u03b1. TNF\u03b1 (10\u00a0ng\/ml) treatment in vitro was found to promote the proliferation of immature Sertoli cells, accompanied with increased NF\u03baB activity and cyclin D1 protein level. The level of Fbxo4 and ubiquitination of cyclin D1 were decreased after TNF\u03b1 treatment. Inhibitor of NF\u03baB or overexpression of Fbxo4 could both reverse the TNF\u03b1\u2010induced proliferation of immature Sertoli cells, meanwhile restore the ubiquitin\u2013proteasome system\u2010dependent degradation of cyclin D1. Overexpression of Fbxo4 could not affect the activation of NF\u03baB caused by TNF\u03b1.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>These results indicate that TNF\u03b1 inhibits the ubiquitination and degradation of cyclin D1 through the NF\u03baB pathway, thereby promoting the proliferation of immature Sertoli cell in vitro and inducing the delay of blood\u2013testis barrier maturation in pubertal rats.<\/jats:p><\/jats:sec>","DOI":"10.1111\/andr.13336","type":"journal-article","created":{"date-parts":[[2022,11,10]],"date-time":"2022-11-10T13:32:31Z","timestamp":1668087151000},"page":"575-590","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["TNF\u03b1 stimulates the proliferation of immature Sertoli cells by attenuating UPS\u2010degradation of cyclin D1 and leads to the delay of BTB maturation in pubertal 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