{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T18:59:00Z","timestamp":1773255540984,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T00:00:00Z","timestamp":1715040000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the National Science Foundation of China","award":["82374173"],"award-info":[{"award-number":["82374173"]}]},{"name":"the National Science Foundation of China","award":["81774034"],"award-info":[{"award-number":["81774034"]}]},{"name":"the National Science Foundation of China","award":["81573725"],"award-info":[{"award-number":["81573725"]}]},{"name":"the National Science Foundation of China","award":["202104a07020020"],"award-info":[{"award-number":["202104a07020020"]}]},{"name":"the National Science Foundation of China","award":["KJ2021A0590"],"award-info":[{"award-number":["KJ2021A0590"]}]},{"name":"the Anhui Provincial Key Research and Development Plan","award":["82374173"],"award-info":[{"award-number":["82374173"]}]},{"name":"the Anhui Provincial Key Research and Development Plan","award":["81774034"],"award-info":[{"award-number":["81774034"]}]},{"name":"the Anhui Provincial Key Research and Development Plan","award":["81573725"],"award-info":[{"award-number":["81573725"]}]},{"name":"the Anhui Provincial Key Research and Development Plan","award":["202104a07020020"],"award-info":[{"award-number":["202104a07020020"]}]},{"name":"the Anhui Provincial Key Research and Development Plan","award":["KJ2021A0590"],"award-info":[{"award-number":["KJ2021A0590"]}]},{"name":"the Key Scientific Research Projects of Anhui Provincial Department of Education","award":["82374173"],"award-info":[{"award-number":["82374173"]}]},{"name":"the Key Scientific Research Projects of Anhui Provincial Department of Education","award":["81774034"],"award-info":[{"award-number":["81774034"]}]},{"name":"the Key Scientific Research Projects of Anhui Provincial Department of Education","award":["81573725"],"award-info":[{"award-number":["81573725"]}]},{"name":"the Key Scientific Research Projects of Anhui Provincial Department of Education","award":["202104a07020020"],"award-info":[{"award-number":["202104a07020020"]}]},{"name":"the Key Scientific Research Projects of Anhui Provincial Department of Education","award":["KJ2021A0590"],"award-info":[{"award-number":["KJ2021A0590"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>The Pulsatilla decoction is a well-known herbal remedy used in clinical settings for treating vulvovaginal candidiasis (VVC). However, the specific mechanism that makes it effective is still unclear. Recent studies have shown that in cases of VVC, neutrophils recruited to the vagina, influenced by heparan sulfate (HS), do not successfully engulf Candida albicans (C. albicans). Instead, they release many inflammatory factors that cause damage to the vaginal mucosa. This study aims to understand the molecular mechanism by which the n-butanol extract of Pulsatilla decoction (BEPD) treats VVC through transcriptomics. High-performance liquid chromatography was used to identify the primary active components of BEPD. A VVC mouse model was induced using an estrogen-dependent method and the mice were treated daily with BEPD (20 mg\/kg, 40 mg\/kg, and 80 mg\/kg) for seven days. The vaginal lavage fluid of the mice was analyzed for various experimental indices, including fungal morphology, fungal burden, degree of neutrophil infiltration, and cytokines. Various assessments were then performed on mouse vaginal tissues, including pathological assessment, immunohistochemistry, immunofluorescence, Western blot (WB), quantitative real-time PCR, and transcriptome assays. Our results showed that BEPD reduced vaginal redness and swelling, decreased white discharge, inhibited C. albicans hyphae formation, reduced neutrophil infiltration and fungal burden, and attenuated vaginal tissue damage compared with the VVC model group. The high-dose BEPD group even restored the damaged vaginal tissue to normal levels. The medium- and high-dose groups of BEPD also significantly reduced the levels of IL-1\u03b2, IL-6, TNF-\u03b1, and LDH. Additionally, transcriptomic results showed that BEPD regulated several chemokine (CXCL1, CXCL3, and CXCL5) and S100 alarmin (S100A8 and S100A9) genes, suggesting that BEPD may treat VVC by affecting chemokine- and alarmin-mediated neutrophil chemotaxis. Finally, we verified that BEPD protects the vaginal mucosa of VVC mice by inhibiting neutrophil recruitment and chemotaxis in an animal model of VVC via the TLR4\/MyD88\/NF-\u03baB pathway. This study provides further evidence to elucidate the mechanism of BEPD treatment of VVC.<\/jats:p>","DOI":"10.3390\/ph17050594","type":"journal-article","created":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T09:03:38Z","timestamp":1715072618000},"page":"594","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Transcriptomics Reveals Effect of Pulsatilla Decoction Butanol Extract in Alleviating Vulvovaginal Candidiasis by Inhibiting Neutrophil Chemotaxis and Activation via TLR4 Signaling"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-3375-9052","authenticated-orcid":false,"given":"Hui","family":"Wu","sequence":"first","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Can","family":"Li","sequence":"additional","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yemei","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mengxiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daqiang","family":"Wu","sequence":"additional","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7540-2969","authenticated-orcid":false,"given":"Jing","family":"Shao","sequence":"additional","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tianming","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Changzhong","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Integrated Traditional and Western Medicine, Anhui University of Chinese Medicine, Hefei 230012, China"},{"name":"Institute of Integrated Traditional Chinese and Western Medicine, Anhui Academy of Chinese Medicine, Hefei 230012, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Shen, L., Zhang, W., Yuan, Y., Zhu, W., and Shang, A. (2022). Vaginal microecological characteristics of women in different physiological and pathological period. Front. Cell. Infect. Microbiol., 12.","DOI":"10.3389\/fcimb.2022.959793"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Kenno, S., Speth, C., Rambach, G., Binder, U., Chatterjee, S., Caramalho, R., Haas, H., Lass-Fl\u00f6rl, C., Shaughnessy, J., and Ram, S. (2019). Candida albicans Factor H Binding Molecule Hgt1p\u2013A Low Glucose-Induced Transmembrane Protein Is Trafficked to the Cell Wall and Impairs Phagocytosis and Killing by Human Neutrophils. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.03319"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"905","DOI":"10.3109\/1040841X.2015.1091805","article-title":"Vulvovaginal candidiasis: Epidemiology, microbiology and risk factors","volume":"42","author":"Ferreira","year":"2016","journal-title":"Crit. Rev. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1111\/myc.13248","article-title":"Guideline: Vulvovaginal candidosis (AWMF 015\/072, level S2k)","volume":"64","author":"Farr","year":"2021","journal-title":"Mycoses"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e339","DOI":"10.1016\/S1473-3099(18)30103-8","article-title":"Global burden of recurrent vulvovaginal candidiasis: A systematic review","volume":"18","author":"Denning","year":"2018","journal-title":"Lancet Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1146\/annurev-pathol-052016-100342","article-title":"Immunity to Commensal Fungi: Detente and Disease","volume":"12","author":"Wheeler","year":"2017","journal-title":"Annu. Rev. Pathol.-Mech. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Naglik, J.R., K\u00f6nig, A., Hube, B., and Gaffen, S.L. (2017). Candida albicans-epithelial interactions and induction of mucosal innate immunity. Curr. Opin. Microbiol., 40.","DOI":"10.1016\/j.mib.2017.10.030"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"17877","DOI":"10.1038\/s41598-017-17649-8","article-title":"NLRP3 inflammasome is a key player in human vulvovaginal disease caused by Candida albicans","volume":"7","author":"Roselletti","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Yano, J., Noverr, M.C., and Fidel, P.L. (2017). Vaginal Heparan Sulfate Linked to Neutrophil Dysfunction in the Acute Inflammatory Response Associated with Experimental Vulvovaginal Candidiasis. mBio, 8.","DOI":"10.1128\/mBio.00211-17"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"281","DOI":"10.2174\/1573405616666200621200047","article-title":"Recent Advances in Vaginal Delivery for the Treatment of Vulvovaginal Candidiasis","volume":"14","author":"Kaur","year":"2021","journal-title":"Curr. Mol. Pharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"814883","DOI":"10.3389\/fphar.2022.814883","article-title":"Berberine Inhibits the Adhesion of Candida albicans to Vaginal Epithelial Cells","volume":"13","author":"Zhao","year":"2022","journal-title":"Front. Pharmacol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"155515","DOI":"10.1016\/j.phymed.2024.155515","article-title":"Effect of Pulsatilla decoction on vulvovaginal candidiasis in mice. Evidences for its mechanisms of action","volume":"128","author":"Hu","year":"2024","journal-title":"Phytomedicine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"155302","DOI":"10.1016\/j.phymed.2023.155302","article-title":"Exploration of the immuno-adjuvant effect and mechanism of Anemoside B4 through network pharmacology and experiment verification","volume":"124","author":"Shen","year":"2024","journal-title":"Phytomedicine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2263","DOI":"10.1007\/s10529-020-02938-6","article-title":"Inhibition of berberine hydrochloride on Candida albicans biofilm formation","volume":"42","author":"Huang","year":"2020","journal-title":"Biotechnol. Lett."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Yu, H., Zhang, S., Li, R., Ma, C., Zhang, Q., Xia, F., Zhou, B., Xie, Z., and Liao, Z. (2024). Berberine alleviates inflammation and suppresses PLA2-COX-2-PGE2-EP2 pathway through targeting gut microbiota in DSS-induced ulcerative colitis. Biochem. Biophys. Res. Commun., 695.","DOI":"10.1016\/j.bbrc.2023.149411"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"117849","DOI":"10.1016\/j.jep.2024.117849","article-title":"Fraxini cortex: Progresses in phytochemistry, pharmacology and ethnomedicinal uses","volume":"325","author":"Zheng","year":"2024","journal-title":"J. Ethnopharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5126","DOI":"10.1128\/IAI.00388-10","article-title":"Epithelial cell-derived S100 calcium-binding proteins as key mediators in the hallmark acute neutrophil response during Candida vaginitis","volume":"78","author":"Yano","year":"2010","journal-title":"Infect. Immun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.cellimm.2017.12.012","article-title":"Knockout of autophagy gene, ATG5 in mice vaginal cells abrogates cytokine response and pathogen clearance during vaginal infection of Candida albicans","volume":"324","author":"Shroff","year":"2018","journal-title":"Cell. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"532","DOI":"10.1128\/IAI.01417-13","article-title":"Fungal morphogenetic pathways are required for the hallmark inflammatory response during Candida albicans vaginitis","volume":"82","author":"Peters","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1128\/IAI.00861-13","article-title":"Vaginal epithelial cell-derived S100 alarmins induced by Candida albicans via pattern recognition receptor interactions are sufficient but not necessary for the acute neutrophil response during experimental vaginal candidiasis","volume":"82","author":"Yano","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7669","DOI":"10.3748\/wjg.v27.i44.7669","article-title":"Calycosin attenuates severe acute pancreatitis-associated acute lung injury by curtailing high mobility group box 1-induced inflammation","volume":"27","author":"Zhu","year":"2021","journal-title":"World J. Gastroenterol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/s12931-022-02307-z","article-title":"The main e-cigarette component vegetable glycerin enhances neutrophil migration and fibrosis in endotoxin-induced lung injury via p38 MAPK activation","volume":"24","author":"Su","year":"2023","journal-title":"Respir. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1940","DOI":"10.1128\/IAI.02895-14","article-title":"Induction of caspase-11 by aspartyl proteinases of Candida albicans and implication in promoting inflammatory response","volume":"83","author":"Gabrielli","year":"2015","journal-title":"Infect. Immun."},{"key":"ref_24","first-page":"183","article-title":"Phagocyte-specific S100 proteins are released from affected mucosa and promote immune responses during inflammatory bowel disease","volume":"216","author":"Foell","year":"2008","journal-title":"J. Pathol. A J. Pathol. Soc. Great Br. Irel."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3233","DOI":"10.4049\/jimmunol.170.6.3233","article-title":"Proinflammatory activities of S100: Proteins S100A8, S100A9, and S100A8\/A9 induce neutrophil chemotaxis and adhesion","volume":"170","author":"Ryckman","year":"2003","journal-title":"J. Immunol."},{"key":"ref_26","first-page":"3664","article-title":"Human epithelial cells establish direct antifungal defense through TLR4-mediated signaling","volume":"117","author":"Weindl","year":"2007","journal-title":"J. Clin. Investig."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1177\/0022034510381264","article-title":"TLR-signaling networks: An integration of adaptor molecules, kinases, and cross-talk","volume":"90","author":"Brown","year":"2011","journal-title":"J. Dent. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"959740","DOI":"10.3389\/fimmu.2022.959740","article-title":"Distinct host immune responses in recurrent vulvovaginal candidiasis and vulvovaginal candidiasis","volume":"13","author":"Ge","year":"2022","journal-title":"Front. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"S392","DOI":"10.1086\/314815","article-title":"Human immunodeficiency virus type 1 infction, mucosalimmunity, and pathogenesis and extramural research programs at the national institutesof health","volume":"179","author":"Kresina","year":"1999","journal-title":"J. Infect. Dis."},{"key":"ref_30","first-page":"345","article-title":"Human hemlgiderived ptics ekibi aimiroiai aciya class of host defense peptides","volume":"791","author":"Liepke","year":"2003","journal-title":"J. Chromatogr."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6859","DOI":"10.2741\/3194","article-title":"Hemocidins: A functional and structural context of human antimicrobial activity peptides","volume":"13","author":"Mak","year":"2008","journal-title":"Front. Biosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"91.e1","DOI":"10.1016\/j.ajog.2018.10.023","article-title":"Vulvovaginal candidiasis: Histologic lesions are primarily polymicrobial and invasive and do not contain biofilms","volume":"220","author":"Swidsinski","year":"2019","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2178","DOI":"10.1182\/blood-2018-11-844530","article-title":"Neutrophils and NETS in modulating acute and chronic inflammation","volume":"133","author":"Castanheira","year":"2019","journal-title":"Blood"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"115763","DOI":"10.1016\/j.jep.2022.115763","article-title":"Jinhua Qinggan granules attenuates acute lung injury by promotion of neutrophil apoptosis and inhibition of TLR4\/MyD88\/NF-\u03baB pathway","volume":"301","author":"Zhu","year":"2023","journal-title":"J. Ethnopharmacol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1016\/j.bbrc.2016.04.024","article-title":"NKT cells mediate the recruitment of neutrophils by stimulating epithelial chemokine secretion during colitis","volume":"474","author":"Huang","year":"2016","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1371\/journal.ppat.1003965","article-title":"Candida Vaginitis: When Opportunism Knocks, the Host Responds","volume":"10","author":"Peters","year":"2014","journal-title":"PLoS Pathog."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Junko, Y., Kolls, J.K., Happel, K.I., Wormley, F., and Wozniak, K.L. (2012). The Acute Neutrophil Response Mediated by S100 Alarmins during Vaginal Candida Infections Is Independent of the Th17-Pathway. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0046311"},{"key":"ref_38","first-page":"26","article-title":"Candidalysin drives epithelial signaling, neutrophil recruitment, and immunopathology at the vaginal mucosa","volume":"78","author":"Richardson","year":"2018","journal-title":"Infect. Immun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"116041","DOI":"10.1016\/j.jep.2022.116041","article-title":"Effects of n-butanol extract of pulsatilla decoction on the NLRP3 inflammasome in macrophages infected with Candida albicans","volume":"304","author":"Hu","year":"2023","journal-title":"J. Ethnopharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e03619","DOI":"10.1016\/j.heliyon.2020.e03619","article-title":"Low prevalence of antifungal resistant Candida africana, in the C. albicans complex causing vulvovaginal candidiasis","volume":"6","author":"Farahyar","year":"2020","journal-title":"Heliyon"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"116869","DOI":"10.1016\/j.jep.2023.116869","article-title":"Longdan Xiegan decoction ameliorates vulvovaginal candidiasis by inhibiting the NLRP3 inflammasome via the Toll-like receptor \/MyD88 pathway","volume":"318","author":"Feng","year":"2024","journal-title":"J. Ethnopharmacol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Xu, B., Yao, M., Liu, Z., Zhang, S., Wang, B., Han, Y., Gao, J., Wu, D., and Wang, X. (2023). Metabolomic Analysis of Stephania tetrandra-Astragalus membranaceus Herbal Pair-Improving Nephrotic Syndrome Identifies Activation of IL-13\/STAT6 Signaling Pathway. Pharmaceuticals, 16.","DOI":"10.3390\/ph16010088"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1038\/s41401-022-00933-3","article-title":"Hederacoside C ameliorates colitis via restoring impaired intestinal barrier through moderating S100A9\/MAPK and neutrophil recruitment inactivation","volume":"44","author":"Zha","year":"2023","journal-title":"Acta Pharmacol. Sin."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/17\/5\/594\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:40:51Z","timestamp":1760107251000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/17\/5\/594"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,7]]},"references-count":43,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["ph17050594"],"URL":"https:\/\/doi.org\/10.3390\/ph17050594","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,7]]}}}