{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T06:22:31Z","timestamp":1791872551986,"version":"4.3.4"},"reference-count":42,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T00:00:00Z","timestamp":1748908800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec024\">\n<jats:title>Background<\/jats:title>\n<jats:p>This study aimed to deepen understanding of the molecular mechanisms and key characteristic genes of non-obstructive azoospermia (NOA).<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec025\">\n<jats:title>Methods<\/jats:title>\n<jats:p>A systematic retrieval method was used to collect the mRNA expression data of NOA and obstructive azoospermia (OA) samples from the GEO database. Data preprocessing, differential gene expression screening, functional annotation, and signal pathway enrichment analysis were conducted using R software. The differences in immune microenvironment between NOA and OA samples were compared through CIBERSORT analysis. LASSO and SVM-RFE, two machine learning algorithms, were applied to select NOA-related characteristic genes. Subsequently, our investigation further identified genes differentially expressed in NOA that are associated with inflammatory responses. NOA samples were clustered based on these inflammation-related genes, while molecular features between different types were explored through pathway enrichment analysis of gene set variation analysis (GSVA). Finally, potential traditional Chinese medicine components targeting these inflammation-related genes were screened from the Chinese medicine database, followed by drug-protein docking simulations.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec026\">\n<jats:title>Results<\/jats:title>\n<jats:p>The study identified 772 DEGs mainly involved in the generation and maturation of sperm. Immune microenvironment analysis revealed significant differences in the infiltration levels of resting NK cells and activated dendritic cells between NOA and OA samples. Eight NOA-related characteristic genes were identified through LASSO and SVM-RFE algorithms. Further analysis revealed that three inflammation-related genes, namely LAMP3, PROK2, and CD14, exhibited significant differential expression in samples of NOA and OA. After clustering of these NOA samples based on the three inflammation-related DEGs, GSVA pathway enrichment analysis revealed molecular features between different NOA subtypes. Finally, potential traditional Chinese medicine components targeting these inflammation-related genes were selected.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec027\">\n<jats:title>Conclusion<\/jats:title>\n<jats:p>This study revealed the key molecular mechanisms and characteristic genes of NOA, especially the role of inflammation-related genes, providing new therapeutic targets and directions for the treatment of NOA.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0324948","type":"journal-article","created":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T17:21:50Z","timestamp":1748971310000},"page":"e0324948","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Analysis of the inflammatory gene expression characteristics and immune microenvironment regulatory mechanisms in the testicular tissue of patients with non-obstructive azoospermia"],"prefix":"10.1371","volume":"20","author":[{"given":"Qiang","family":"Ling","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingqi","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunhua","family":"Liao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-4736-8478","authenticated-orcid":true,"given":"Guijian","family":"Pang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,6,3]]},"reference":[{"issue":"1","key":"pone.0324948.ref001","doi-asserted-by":"crossref","first-page":"5683","DOI":"10.1038\/s41467-020-19414-4","article-title":"Single-cell analysis of developing and azoospermia human testicles reveals central role of Sertoli cells","volume":"11","author":"L Zhao","year":"2020","journal-title":"Nat Commun"},{"issue":"6","key":"pone.0324948.ref002","doi-asserted-by":"crossref","first-page":"763","DOI":"10.1093\/humupd\/dmac024","article-title":"Meiotic recombination: insights into its mechanisms and its role in human reproduction with a special focus on non-obstructive azoospermia","volume":"28","author":"C Xie","year":"2022","journal-title":"Hum Reprod Update"},{"issue":"20","key":"pone.0324948.ref003","doi-asserted-by":"crossref","DOI":"10.3390\/ijms232012570","article-title":"Whole exome sequencing and in silico analysis of human sertoli in patients with non-obstructive azoospermia","volume":"23","author":"H Azizi","year":"2022","journal-title":"Int J Mol Sci"},{"key":"pone.0324948.ref004","doi-asserted-by":"crossref","first-page":"658432","DOI":"10.3389\/fimmu.2021.658432","article-title":"Impacts of immunometabolism on male reproduction","volume":"12","author":"L Ye","year":"2021","journal-title":"Front Immunol"},{"key":"pone.0324948.ref005","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.gene.2019.04.028","article-title":"Investigation of CD133 and CD24 as candidate azoospermia markers and their relationship with spermatogenesis defects","volume":"706","author":"Y Yukselten","year":"2019","journal-title":"Gene"},{"issue":"5","key":"pone.0324948.ref006","doi-asserted-by":"crossref","first-page":"7052","DOI":"10.18632\/aging.202559","article-title":"Infertility network and hub genes for nonobstructive azoospermia utilizing integrative analysis","volume":"13","author":"B Han","year":"2021","journal-title":"Aging (Albany NY)"},{"issue":"3","key":"pone.0324948.ref007","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1007\/s12020-016-1178-0","article-title":"Delayed treatment of undescended testes may promote hypogonadism and infertility","volume":"55","author":"J Rohayem","year":"2017","journal-title":"Endocrine"},{"key":"pone.0324948.ref008","doi-asserted-by":"crossref","first-page":"897029","DOI":"10.3389\/fendo.2022.897029","article-title":"Mechanism of inflammatory associated impairment of sperm function, spermatogenesis and steroidogenesis","volume":"13","author":"H Hasan","year":"2022","journal-title":"Front Endocrinol"},{"issue":"1","key":"pone.0324948.ref009","doi-asserted-by":"crossref","DOI":"10.3390\/ijms22010057","article-title":"T lymphocytes and testicular immunity: a new insight into immune regulation in testes","volume":"22","author":"J Gong","year":"2020","journal-title":"Int J Mol Sci"},{"key":"pone.0324948.ref010","doi-asserted-by":"crossref","first-page":"7","DOI":"10.12703\/r\/10-7","article-title":"Non-obstructive azoospermia: current and future perspectives","volume":"10","author":"T Tharakan","year":"2021","journal-title":"Fac Rev"},{"issue":"7","key":"pone.0324948.ref011","doi-asserted-by":"crossref","DOI":"10.3390\/jcm10071400","article-title":"Reproductive chances of men with azoospermia due to spermatogenic dysfunction","volume":"10","author":"C Kang","year":"2021","journal-title":"J Clin Med"},{"issue":"18","key":"pone.0324948.ref012","doi-asserted-by":"crossref","first-page":"4029","DOI":"10.1242\/dev.02003","article-title":"RNF17, a component of the mammalian germ cell nuage, is essential for spermiogenesis","volume":"132","author":"J Pan","year":"2005","journal-title":"Development"},{"issue":"9397","key":"pone.0324948.ref013","doi-asserted-by":"crossref","first-page":"1714","DOI":"10.1016\/S0140-6736(03)14845-3","article-title":"Azoospermia in patients heterozygous for a mutation in SYCP3","volume":"362","author":"T Miyamoto","year":"2003","journal-title":"Lancet"},{"issue":"4","key":"pone.0324948.ref014","doi-asserted-by":"crossref","first-page":"1245","DOI":"10.1111\/jcmm.17180","article-title":"Novel exon mutation in SYCE1 gene is associated with non-obstructive azoospermia","volume":"26","author":"K Feng","year":"2022","journal-title":"J Cell Mol Med"},{"issue":"6","key":"pone.0324948.ref015","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1007\/s10815-015-0445-y","article-title":"Deleterious mutation in SYCE1 is associated with non-obstructive azoospermia","volume":"32","author":"E Maor-Sagie","year":"2015","journal-title":"J Assist Reprod Genet"},{"issue":"2","key":"pone.0324948.ref016","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1007\/s10815-019-01660-1","article-title":"The second mutation of SYCE1 gene associated with autosomal recessive nonobstructive azoospermia","volume":"37","author":"M Pashaei","year":"2020","journal-title":"J Assist Reprod Genet"},{"issue":"8","key":"pone.0324948.ref017","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1038\/35087082","article-title":"Mammalian STAG3 is a cohesin specific to sister chromatid arms in meiosis I","volume":"3","author":"I Prieto","year":"2001","journal-title":"Nat Cell Biol"},{"issue":"9","key":"pone.0324948.ref018","doi-asserted-by":"crossref","first-page":"2316","DOI":"10.1093\/humrep\/deq180","article-title":"Dynamics of cohesin proteins REC8, STAG3, SMC1 beta and SMC3 are consistent with a role in sister chromatid cohesion during meiosis in human oocytes","volume":"25","author":"R Garcia-Cruz","year":"2010","journal-title":"Hum Reprod"},{"issue":"3","key":"pone.0324948.ref019","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1096\/fasebj.14.3.581","article-title":"STAG3, a novel gene encoding a protein involved in meiotic chromosome pairing and location of STAG3-related genes flanking the Williams-Beuren syndrome deletion","volume":"14","author":"N Pezzi","year":"2000","journal-title":"FASEB J"},{"issue":"1","key":"pone.0324948.ref020","doi-asserted-by":"crossref","first-page":"10077","DOI":"10.1038\/s41598-021-89559-9","article-title":"Analysis of STAG3 variants in Chinese non-obstructive azoospermia patients with germ cell maturation arrest","volume":"11","author":"W Liu","year":"2021","journal-title":"Sci Rep"},{"issue":"11","key":"pone.0324948.ref021","first-page":"2112","article-title":"Mutations in the stromal antigen 3 (STAG3) gene cause male infertility due to meiotic arrest","volume":"34","author":"N vanderBijl","year":"2019","journal-title":"Hum Reprod"},{"issue":"1","key":"pone.0324948.ref022","doi-asserted-by":"crossref","first-page":"106","DOI":"10.4103\/aja.aja_28_19","article-title":"The association of stromal antigen 3 (STAG3) sequence variations with spermatogenic impairment in the male Korean population","volume":"22","author":"Y Nam","year":"2020","journal-title":"Asian J Androl"},{"issue":"9","key":"pone.0324948.ref023","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1093\/molehr\/gaaa050","article-title":"STAG3 homozygous missense variant causes primary ovarian insufficiency and male non-obstructive azoospermia","volume":"26","author":"S Jaillard","year":"2020","journal-title":"Mol Hum Reprod"},{"issue":"11","key":"pone.0324948.ref024","doi-asserted-by":"crossref","first-page":"1243","DOI":"10.1002\/embj.201387329","article-title":"STAG3-mediated stabilization of REC8 cohesin complexes promotes chromosome synapsis during meiosis","volume":"33","author":"T Fukuda","year":"2014","journal-title":"EMBO J"},{"issue":"6","key":"pone.0324948.ref025","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1093\/molehr\/7.6.513","article-title":"Teratozoospermia in mice lacking the transition protein 2 (Tnp2)","volume":"7","author":"I Adham","year":"2001","journal-title":"Mol Hum Reprod"},{"issue":"4","key":"pone.0324948.ref026","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1111\/j.1439-0272.2007.00775.x","article-title":"Transition protein 1 mRNA expression is not related to pregnancy rate in azoospermic men undergoing TESE-ICSI","volume":"39","author":"V Mitchell","year":"2007","journal-title":"Andrologia"},{"issue":"21","key":"pone.0324948.ref027","doi-asserted-by":"crossref","first-page":"7243","DOI":"10.1128\/MCB.21.21.7243-7255.2001","article-title":"Targeted disruption of the transition protein 2 gene affects sperm chromatin structure and reduces fertility in mice","volume":"21","author":"M Zhao","year":"2001","journal-title":"Mol Cell Biol"},{"issue":"1","key":"pone.0324948.ref028","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1002\/mrd.22440","article-title":"Quantification of sperm specific mRNA transcripts (PRM1, PRM2, and TNP2) in teratozoospermia and normozoospermia: New correlations between mRNA content and morphology of sperm","volume":"82","author":"E Savadi-Shiraz","year":"2015","journal-title":"Mol Reprod Dev"},{"key":"pone.0324948.ref029","doi-asserted-by":"crossref","first-page":"865967","DOI":"10.3389\/fvets.2022.865967","article-title":"Chronic immune-mediated orchitis is the major cause of acquired non-obstructive azoospermia in dogs","volume":"9","author":"S Goericke-Pesch","year":"2022","journal-title":"Front Vet Sci"},{"issue":"5","key":"pone.0324948.ref030","doi-asserted-by":"crossref","DOI":"10.1111\/aji.13481","article-title":"Evaluation of immune status in testis and macrophage polarization associated with testicular damage in patients with nonobstructive azoospermia","volume":"86","author":"W Zheng","year":"2021","journal-title":"Am J Reprod Immunol"},{"issue":"11","key":"pone.0324948.ref031","doi-asserted-by":"crossref","DOI":"10.1111\/and.13120","article-title":"Exploring the role of antigen presenting cells in male genital tract","volume":"50","author":"V Guazzone","year":"2018","journal-title":"Andrologia"},{"issue":"5","key":"pone.0324948.ref032","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1530\/REP-10-0493","article-title":"A dense network of dendritic cells populates the murine epididymis","volume":"141","author":"N Da Silva","year":"2011","journal-title":"Reprod"},{"issue":"2","key":"pone.0324948.ref033","doi-asserted-by":"crossref","first-page":"175","DOI":"10.5534\/wjmh.180069","article-title":"Traditional Chinese Medicine as a remedy for male infertility: a review","volume":"37","author":"SH Zhou","year":"2019","journal-title":"World J Mens Health"},{"issue":"1","key":"pone.0324948.ref034","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1038\/s41419-021-04435-8","article-title":"Epimedium protects against dyszoospermia in mice with Pex3 knockout by exerting antioxidant effects and regulating the expression level of P16","volume":"13","author":"H Zhao","year":"2022","journal-title":"Cell Death Dis"},{"key":"pone.0324948.ref035","doi-asserted-by":"crossref","first-page":"111514","DOI":"10.1016\/j.biopha.2021.111514","article-title":"Xiaokang liuwei dihuang decoction ameliorates the immune infertility of male rats induced by lipopolysaccharide through regulating the levels of sex hormones, reactive oxygen species, pro-apoptotic and immune factors","volume":"139","author":"X Sun","year":"2021","journal-title":"Biomed Pharmacother"},{"key":"pone.0324948.ref036","doi-asserted-by":"crossref","first-page":"794448","DOI":"10.3389\/fphar.2022.794448","article-title":"An integrative approach to uncover the components, mechanisms, and functions of traditional Chinese medicine prescriptions on male infertility","volume":"13","author":"X Bai","year":"2022","journal-title":"Front Pharmacol"},{"key":"pone.0324948.ref037","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.phrs.2018.09.002","article-title":"Resolving neuroinflammation, the therapeutic potential of the anti-malaria drug family of artemisinin","volume":"136","author":"Z Shi","year":"2018","journal-title":"Pharmacol Res"},{"issue":"18","key":"pone.0324948.ref038","doi-asserted-by":"crossref","DOI":"10.3390\/molecules26185698","article-title":"Molecular basis of artemisinin derivatives inhibition of myeloid differentiation protein 2 by combined in silico and experimental study","volume":"26","author":"S Qiao","year":"2021","journal-title":"Molecules"},{"issue":"4","key":"pone.0324948.ref039","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1007\/s00011-019-01216-0","article-title":"Artemisinin and its derivatives: a potential therapeutic approach for oral lichen planus","volume":"68","author":"R-J Ma","year":"2019","journal-title":"Inflamm Res"},{"key":"pone.0324948.ref040","doi-asserted-by":"crossref","first-page":"107284","DOI":"10.1016\/j.intimp.2020.107284","article-title":"Artemisinin ameliorates intestinal inflammation by skewing macrophages to the M2 phenotype and inhibiting epithelial-mesenchymal transition","volume":"91","author":"M Huai","year":"2021","journal-title":"Int Immunopharmacol"},{"issue":"1","key":"pone.0324948.ref041","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1080\/08923973.2016.1267744","article-title":"Artemisinin inhibits inflammatory response via regulating NF-\u03baB and MAPK signaling pathways","volume":"39","author":"KS Wang","year":"2017","journal-title":"Immunopharmacol Immunotoxicol"},{"issue":"21","key":"pone.0324948.ref042","article-title":"The effect of artemisinin on inflammation-associated lymphangiogenesis in experimental acute colitis","volume":"21","author":"A Lee","year":"2020","journal-title":"Int J Mol Sci"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0324948","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T17:22:08Z","timestamp":1748971328000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0324948"}},"subtitle":[],"editor":[{"given":"Sule","family":"Dogan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,6,3]]},"references-count":42,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6,3]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0324948","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,3]]}}}