{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T11:26:46Z","timestamp":1780918006573,"version":"3.54.1"},"reference-count":45,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T00:00:00Z","timestamp":1772496000000},"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. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Polycystic ovary syndrome (PCOS) is associated with chronic low-grade inflammation and insulin signaling dysregulation. Gui Shen Wan (GSW), a traditional Chinese medicine formula, has been used empirically for ovarian dysfunction, yet its molecular basis remains incompletely defined. This study aimed to delineate the mechanism by which GSW modulates inflammation-linked insulin signaling in a PCOS-relevant granulosa cell model under metabolic stress, with a focus on the TNF-\u03b1\/PI3K\/Akt axis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Network pharmacology based on serum-absorbed constituents identified by UPLC\u2013MS\/MS was integrated with\n                      <jats:italic>in vitro<\/jats:italic>\n                      validation using dexamethasone- and insulin\u2013challenged human KGN cells to model selected PCOS-relevant cellular phenotypes. The effects of GSW-medicated serum on cell viability, apoptosis, hormone-associated readouts, and inflammatory cytokine production were assessed. PI3K\/Akt signaling was examined by Western blotting, and a recombinant TNF-\u03b1 rescue experiment was performed to probe mechanistic dependence. The effects of candidate constituents (embelin and nobiletin) were further evaluated.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Network pharmacology highlighted TNF and PI3K\/Akt signaling as key pathways. In the KGN metabolic stress model, GSW-medicated serum dose-dependently improved cell viability, reduced apoptosis, and attenuated inflammatory cytokine output (TNF-\u03b1 and IL-6), accompanied by increased phosphorylation of PI3K and Akt. Recombinant TNF-\u03b1 markedly diminished the protective and signaling-activating effects of GSW, supporting a TNF-\u03b1\u2013linked mechanism. Embelin and nobiletin reproduced key anti-inflammatory and signaling effects, and their co-application produced an enhanced combined effect at the tested concentrations.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      These findings suggest that GSW mitigates PCOS-like granulosa cell dysfunction under metabolic stress by suppressing TNF-\u03b1\u2013associated inflammatory signaling, thereby relieving inhibition of the PI3K\/Akt pathway. Given the\n                      <jats:italic>in vitro<\/jats:italic>\n                      scope and the medicated-serum approach, the results should be interpreted as mechanistic insight rather than direct evidence of clinical efficacy, and they provide a rationale for subsequent\n                      <jats:italic>in vivo<\/jats:italic>\n                      validation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1739230","type":"journal-article","created":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T05:22:32Z","timestamp":1772515352000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Gui Shen Wan ameliorates PCOS-like cellular phenotypes by suppressing TNF-\u03b1-mediated inflammation and restoring the PI3K\/Akt signaling pathway"],"prefix":"10.3389","volume":"17","author":[{"given":"Yan","family":"Lu","sequence":"first","affiliation":[{"name":"The Affiliated Hospital Of Jiangxi University Of CM","place":["Nanchang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingtong","family":"Li","sequence":"additional","affiliation":[{"name":"The Second School of Clinical Medicine, Guangzhou University of Chinese Medicine","place":["Guangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia","family":"Fang","sequence":"additional","affiliation":[{"name":"The Affiliated Hospital Of Jiangxi University Of CM","place":["Nanchang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjuan","family":"Ju","sequence":"additional","affiliation":[{"name":"The Affiliated Hospital Of Jiangxi University Of CM","place":["Nanchang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanfang","family":"Yan","sequence":"additional","affiliation":[{"name":"The Affiliated Hospital Of Jiangxi University Of CM","place":["Nanchang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feihua","family":"Wu","sequence":"additional","affiliation":[{"name":"The Affiliated Hospital Of Jiangxi University Of CM","place":["Nanchang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,3,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1007\/s00404-020-05951-2","article-title":"Association of metabolic and inflammatory markers with polycystic ovarian syndrome (PCOS): an update","volume":"303","author":"Abraham Gnanadass","year":"2021","journal-title":"Arch Gynecol Obstet"},{"key":"B2","doi-asserted-by":"publisher","first-page":"121069","DOI":"10.1016\/j.lfs.2022.121069","article-title":"Polycystic ovary syndrome (PCOS) increases the risk of subsequent gestational diabetes mellitus (GDM): A novel therapeutic perspective","volume":"310","author":"Choudhury","year":"2022","journal-title":"Life Sci"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2024.1358755","article-title":"PCOS stratification for precision diagnostics and treatment","volume":"12","author":"Joshi","year":"2024","journal-title":"Front Cell Dev Biol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1007\/s10815-022-02625-7","article-title":"A brief insight into the etiology, genetics, and immunology of polycystic ovarian syndrome (PCOS)","volume":"39","author":"Siddiqui","year":"2022","journal-title":"J Assist Reprod Genet"},{"key":"B5","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/j.jsbmb.2018.04.008","article-title":"Polycystic ovary syndrome (PCOS), an inflammatory, systemic, lifestyle endocrinopathy","volume":"182","author":"Patel","year":"2018","journal-title":"J Steroid Biochem Mol Biol"},{"key":"B6","doi-asserted-by":"publisher","first-page":"3789","DOI":"10.3390\/ijms22073789","article-title":"Chronic low grade inflammation in pathogenesis of PCOS","volume":"22","author":"Rudnicka","year":"2021","journal-title":"Int J Mol Sci"},{"key":"B7","doi-asserted-by":"publisher","first-page":"4110","DOI":"10.3390\/ijms23084110","article-title":"Controversies in the pathogenesis, diagnosis and treatment of PCOS: focus on insulin resistance, inflammation, and hyperandrogenism","volume":"23","author":"Armanini","year":"2022","journal-title":"Int J Mol Sci"},{"key":"B8","doi-asserted-by":"publisher","first-page":"2160569","DOI":"10.1080\/13510002.2022.2160569","article-title":"Novel insights into the synergistic effects of selenium nanoparticles and metformin treatment of letrozole - induced polycystic ovarian syndrome: targeting PI3K\/Akt signalling pathway, redox status and mitochondrial dysfunction in ovarian tissue","volume":"28","author":"Rabah","year":"2023","journal-title":"Redox Rep"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.2478\/enr-2018-0026","article-title":"Polycystic Ovary Syndrome as a systemic disease with multiple molecular pathways: a narrative review","volume":"52","author":"Carvalho","year":"2018","journal-title":"Endocr Regul"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1210\/jcem","article-title":"TNF-alpha and hyperandrogenism: a clinical, biochemical, and molecular genetic study","volume":"86","author":"Escobar-Morreale","year":"2001","journal-title":"J Clin Endocrinol Metab"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1089\/gtmb.2014.0151","article-title":"Tumor necrosis factor-alpha and polycystic ovarian syndrome: a clinical, biochemical, and molecular genetic study","volume":"18","author":"Thathapudi","year":"2014","journal-title":"Genet Test Mol Biomarkers"},{"key":"B12","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1186\/s12958-020-00677-x","article-title":"Growth hormone activates PI3K\/Akt signaling and inhibits ROS accumulation and apoptosis in granulosa cells of patients with polycystic ovary syndrome","volume":"18","author":"Gong","year":"2020","journal-title":"Reprod Biol Endocrinol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2022.1091147","article-title":"Insulin resistance, autophagy and apoptosis in patients with polycystic ovary syndrome: Association with PI3K signaling pathway","volume":"13","author":"Tong","year":"2022","journal-title":"Front Endocrinol (Lausanne)"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2023.1191759","article-title":"Signaling pathways and targeted therapeutic strategies for polycystic ovary syndrome","volume":"14","author":"Wang","year":"2023","journal-title":"Front Endocrinol (Lausanne)"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1210\/clinem\/dgad465","article-title":"Metformin and combined oral contraceptive pills in the management of polycystic ovary syndrome: A systematic review and meta-analysis","volume":"109","author":"Melin","year":"2024","journal-title":"J Clin Endocrinol Metab"},{"key":"B16","doi-asserted-by":"publisher","first-page":"CD005552","DOI":"10.1002\/14651858.CD005552.pub3","article-title":"Metformin versus the combined oral contraceptive pill for hirsutism, acne, and menstrual pattern in polycystic ovary syndrome","volume":"8","author":"Fraison","year":"2020","journal-title":"Cochrane Database Syst Rev"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2012.08.026","article-title":"Topical treatment with Tong-Luo-San-Jie gel alleviates bone cancer pain in rats","volume":"143","author":"Wang","year":"2012","journal-title":"J Ethnopharmacol"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2024.1505958","article-title":"The depletion of gut microbiome impairs the beneficial effect of Gui-Shen-Wan in restoring mice ovarian function and associated protein expression of ovarian tissues","volume":"14","author":"Huang","year":"2024","journal-title":"Front Cell Infect Microbiol"},{"key":"B19","doi-asserted-by":"publisher","first-page":"112177","DOI":"10.1016\/j.jep.2019.112177","article-title":"Guizhi Fuling Wan for uterine fibroids: A systematic review of in vivo studies","volume":"245","author":"Li","year":"2019","journal-title":"J Ethnopharmacol"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2020.00575","article-title":"Chinese herbal medicine, ameliorates insulin sensitivity in PCOS model rats with insulin resistance via remodeling intestinal homeostasis","volume":"11","author":"Zhu","year":"2020","journal-title":"Front Endocrinol (Lausanne)"},{"key":"B21","doi-asserted-by":"publisher","first-page":"2210232","DOI":"10.1080\/09513590.2023.2210232","article-title":"Guizhi Fuling Wan inhibits autophagy of granulosa cells in polycystic ovary syndrome mice via H19\/miR-29b-3p","volume":"39","author":"Wu","year":"2023","journal-title":"Gynecol Endocrinol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0308168","article-title":"Unique features of KGN granulosa-like tumour cells in the regulation of steroidogenic and antioxidant genes","volume":"19","author":"Tang","year":"2024","journal-title":"PLoS One"},{"key":"B23","doi-asserted-by":"publisher","first-page":"398","DOI":"10.3390\/cells12030398","article-title":"DEHP Decreases Steroidogenesis through the cAMP and ERK1\/2 Signaling Pathways in FSH-Stimulated Human Granulosa Cells","volume":"12","author":"Tesic","year":"2023","journal-title":"Cells"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jafc.9b07908","article-title":"Nobiletin triggers reactive oxygen species-mediated pyroptosis through regulating autophagy in ovarian cancer cells","volume":"68","author":"Zhang","year":"2020","journal-title":"J Agric Food Chem"},{"key":"B25","doi-asserted-by":"publisher","first-page":"4699","DOI":"10.3390\/ijms22094699","article-title":"Procaspase-activating compound-1 synergizes with TRAIL to induce apoptosis in established granulosa cell tumor cell line (KGN) and explanted patient granulosa cell tumor cells in vitro","volume":"22","author":"Crosley","year":"2021","journal-title":"Int J Mol Sci"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.35629\/4494-1002566574","article-title":"Polycystic ovary syndrome (PCOS): A complex endocrine disorder","volume":"10","author":"Zalwade","year":"2025","journal-title":"Int J Pharm Res Appl"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1080\/07315724.2019.1657515","article-title":"A new approach to polycystic ovary syndrome: the gut microbiota","volume":"39","author":"Yurtdas","year":"2020","journal-title":"J Am Coll Nutr"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.3389\/fpsyt.2022.1001484","article-title":"Depression in polycystic ovary syndrome: Focusing on pathogenesis and treatment","volume":"13","author":"Xing","year":"2022","journal-title":"Front Psychiatry"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1080\/09513590.2021.1958311","article-title":"Progranulin and tumor necrosis factor-alpha in lean polycystic ovary syndrome patients","volume":"37","author":"Uzdogan","year":"2021","journal-title":"Gynecol Endocrinol"},{"key":"B30","doi-asserted-by":"publisher","first-page":"9620423","DOI":"10.1155\/2022\/9620423","article-title":"Plasma diaphanous related formin 1 levels are associated with altered glucose metabolism and insulin resistance in patients with polycystic ovary syndrome: A case control study","volume":"2022","author":"Li","year":"2022","journal-title":"Mediators Inflammation"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.23741","article-title":"CPEB1 induces autophagy and promotes apoptosis in ovarian granulosa cells of polycystic ovary syndrome","volume":"91","author":"Xuan","year":"2024","journal-title":"Mol Reprod Dev"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.2147\/DDDT.S456656","article-title":"Mechanism of YJKL decoction in treating of PCOS infertility by integrative approach of network pharmacology and experimental verification","volume":"18","author":"Zhang","year":"2024","journal-title":"Drug Des Devel Ther"},{"key":"B33","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1186\/s13048-022-00964-8","article-title":"Investigation of the mechanisms and experimental verification of Cuscuta-Salvia in the treatment of polycystic ovary syndrome (PCOS) via network pharmacology","volume":"15","author":"Zhang","year":"2022","journal-title":"J Ovarian Res"},{"key":"B34","doi-asserted-by":"publisher","first-page":"112584","DOI":"10.1016\/j.jep.2020.112584","article-title":"Liu Shen Wan inhibits influenza a virus and excessive virus-induced inflammatory response via suppression of TLR4\/NF-kappaB signaling pathway in vitro and in vivo","volume":"252","author":"Ma","year":"2020","journal-title":"J Ethnopharmacol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2018.08.028","article-title":"Jin Gui Shen Qi Wan, a traditional Chinese medicine, alleviated allergic airway hypersensitivity and inflammatory cell infiltration in a chronic asthma mouse model","volume":"227","author":"Kao","year":"2018","journal-title":"J Ethnopharmacol"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0217495","article-title":"ETA-mediated anti-TNF-alpha therapy ameliorates the phenotype of PCOS model induced by letrozole","volume":"14","author":"Lang","year":"2019","journal-title":"PLoS One"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1007\/s10815-022-02396-1","article-title":"An integrated in silico analysis highlighted angiogenesis regulating miRNA-mRNA network in PCOS pathophysiology","volume":"39","author":"Patil","year":"2022","journal-title":"J Assist Reprod Genet"},{"key":"B38","doi-asserted-by":"publisher","first-page":"119020","DOI":"10.1016\/j.jep.2024.119020","article-title":"Effect of Rhei Radix Et Rhizome on treatment of polycystic ovary syndrome by regulating PI3K\/AKT pathway and targeting EGFR\/ALB in rats","volume":"338","author":"Yang","year":"2025","journal-title":"J Ethnopharmacol"},{"key":"B39","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/s12958-020-00684-y","article-title":"Berberine exerts a protective effect on rats with polycystic ovary syndrome by inhibiting the inflammatory response and cell apoptosis","volume":"19","author":"Shen","year":"2021","journal-title":"Reprod Biol Endocrinol"},{"key":"B40","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1530\/REP-20-0643","article-title":"Melatonin ameliorates ovarian dysfunction by regulating autophagy in PCOS via the PI3K-Akt pathway","volume":"162","author":"Xie","year":"2021","journal-title":"Reproduction"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1152\/ajpendo.00034.2021","article-title":"Luteolin alleviates polycystic ovary syndrome in rats by resolving insulin resistance and oxidative stress","volume":"320","author":"Huang","year":"2021","journal-title":"Am J Physiol Endocrinol Metab"},{"key":"B42","doi-asserted-by":"publisher","first-page":"100490","DOI":"10.1016\/j.obmed.2023.100490","article-title":"Quercetin as a noteworthy treatment for polycystic ovary syndrome: An evidence-based review from molecular basis mechanism to clinical perspective","volume":"39","author":"Habiburrahman","year":"2023","journal-title":"Obes Med"},{"key":"B43","doi-asserted-by":"publisher","first-page":"116418","DOI":"10.1016\/j.biopha.2024.116418","article-title":"Therapeutic effects and molecular mechanisms of quercetin in gynecological disorders","volume":"173","author":"Jian","year":"2024","journal-title":"BioMed Pharmacother"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.202001258","article-title":"Poly-pharmacokinetic strategy represented the synergy effects of bioactive compounds in a traditional Chinese medicine formula, Si Shen Wan and its separated recipes to normal and colitis rats","volume":"44","author":"Liu","year":"2021","journal-title":"J Sep Sci"},{"key":"B45","doi-asserted-by":"publisher","first-page":"154732","DOI":"10.1016\/j.phymed.2023.154732","article-title":"Active post-transcriptional regulation and ACLY-mediated acetyl-CoA synthesis as a pivotal target of Shuang-Huang-Sheng-Bai formula for lung adenocarcinoma treatment","volume":"113","author":"Liu","year":"2023","journal-title":"Phytomedicine"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1739230\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T05:22:33Z","timestamp":1772515353000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1739230\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,3]]},"references-count":45,"alternative-id":["10.3389\/fimmu.2026.1739230"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2026.1739230","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,3]]},"article-number":"1739230"}}