{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T20:40:27Z","timestamp":1781901627253,"version":"3.54.5"},"reference-count":48,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,11,8]],"date-time":"2021-11-08T00:00:00Z","timestamp":1636329600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004735","name":"Natural Science Foundation of Hunan Province","doi-asserted-by":"publisher","award":["2020JJ8109"],"award-info":[{"award-number":["2020JJ8109"]}],"id":[{"id":"10.13039\/501100004735","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:p>Chewing areca nut (betel quid) is strongly associated with oral submucous fibrosis (OSF), a pre-cancerous lesion. Among the areca alkaloids, arecoline is the main agent responsible for fibroblast proliferation; however, the specific molecular mechanism of arecoline affecting the OSF remains unclear. The present study revealed that arecoline treatment significantly enhanced Transforming growth factor-\u03b2 (TGF-\u03b2)-induced buccal mucosal fibroblast (BMF) activation and fibrotic changes. Arecoline interacts with phosphodiesterase 4A (PDE4A) to exert its effects through modulating PDE4A activity but not PDE4A expression. PDE4A silence reversed the effects of arecoline on TGF-\u03b2-induced BMFs activation and fibrotic changes. Moreover, the exchange protein directly activated by cAMP 1 (Epac1)-selective Cyclic adenosine 3\u2032,5\u2032-monophosphate (cAMP) analog (8-Me-cAMP) but not the protein kinase A (PKA)-selective cAMP analog (N6-cAMP) remarkably suppressed \u03b1-smooth muscle actin(\u03b1-SMA) and Collagen Type I Alpha 1 Chain (Col1A1) protein levels in response to TGF-\u03b21 and arecoline co-treatment, indicating that cAMP-Epac1 but not cAMP-PKA signaling is involved in arecoline functions on TGF-\u03b21-induced BMFs activation. In conclusion, arecoline promotes TGF-\u03b21-induced BMFs activation through enhancing PDE4A activity and the cAMP-Epac1 signaling pathway during OSF. This novel mechanism might provide more powerful strategies for OSF treatment, requiring further <jats:italic>in vivo<\/jats:italic> and clinical investigation.<\/jats:p>","DOI":"10.3389\/fphar.2021.722040","type":"journal-article","created":{"date-parts":[[2021,11,8]],"date-time":"2021-11-08T05:36:05Z","timestamp":1636349765000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":17,"title":["Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-\u03b2-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway"],"prefix":"10.3389","volume":"12","author":[{"given":"Bo","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lihua","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunsheng","family":"Shao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingsi","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liangjian","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,11,8]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"5882","DOI":"10.1128\/mcb.19.9.5882","article-title":"Protein Kinase A-dependent and -independent Signaling Pathways Contribute to Cyclic AMP-Stimulated Proliferation","volume":"19","author":"Cass","year":"1999","journal-title":"Mol. Cel Biol"},{"key":"B2","doi-asserted-by":"publisher","first-page":"649","DOI":"10.1016\/j.neo.2017.05.002","article-title":"UALCAN: A Portal for Facilitating Tumor Subgroup Gene Expression and Survival Analyses","volume":"19","author":"Chandrashekar","year":"2017","journal-title":"Neoplasia"},{"key":"B3","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1111\/j.1600-0714.1998.tb02096.x","article-title":"Cytotoxic and Non-genotoxic Effects of Arecoline on Human Buccal Fibroblasts In Vitro","volume":"27","author":"Chang","year":"1998","journal-title":"J. Oral Pathol. Med."},{"key":"B4","doi-asserted-by":"publisher","first-page":"698","DOI":"10.1111\/jcmm.12219","article-title":"Arecoline-induced Myofibroblast Transdifferentiation from Human Buccal Mucosal Fibroblasts Is Mediated by ZEB1","volume":"18","author":"Chang","year":"2014","journal-title":"J. Cel Mol Med"},{"key":"B5","doi-asserted-by":"publisher","first-page":"425","DOI":"10.1016\/s1368-8375(01)00083-5","article-title":"Elevated Vimentin Expression in Buccal Mucosal Fibroblasts by Arecoline In Vitro as a Possible Pathogenesis for Oral Submucous Fibrosis","volume":"38","author":"Chang","year":"2002","journal-title":"Oral Oncol."},{"key":"B6","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1152\/physrev.00007.2018","article-title":"Structure, Gating, and Regulation of the Cftr Anion Channel","volume":"99","author":"Csan\u00e1dy","year":"2019","journal-title":"Physiol. Rev."},{"key":"B7","doi-asserted-by":"publisher","first-page":"1846","DOI":"10.1093\/bioinformatics\/btm254","article-title":"GEOquery: a Bridge between the Gene Expression Omnibus (GEO) and BioConductor","volume":"23","author":"Davis","year":"2007","journal-title":"Bioinformatics"},{"key":"B8","doi-asserted-by":"publisher","first-page":"637","DOI":"10.1089\/ars.2017.7041","article-title":"PDE\/cAMP\/Epac\/C\/EBP-\u03b2 Signaling Cascade Regulates Mitochondria Biogenesis of Tubular Epithelial Cells in Renal Fibrosis","volume":"29","author":"Ding","year":"2018","journal-title":"Antioxid. Redox Signal."},{"key":"B9","doi-asserted-by":"publisher","first-page":"6717","DOI":"10.1128\/mcb.17.11.6717","article-title":"Activation of Cyclic AMP-dependent Kinase Is Required but May Not Be Sufficient to Mimic Cyclic AMP-dependent DNA Synthesis and Thyroglobulin Expression in Dog Thyroid Cells","volume":"17","author":"Dremier","year":"1997","journal-title":"Mol. Cel Biol"},{"key":"B10","doi-asserted-by":"publisher","first-page":"192","DOI":"10.1016\/j.oooo.2015.12.018","article-title":"Oral Submucous Fibrosis: Review on Mechanisms of Malignant Transformation","volume":"122","author":"Ekanayaka","year":"2016","journal-title":"Oral Surg. Oral Med. Oral Pathol. Oral Radiol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/j.lfs.2019.03.014","article-title":"Phosphodiestrase-1 and 4 Inhibitors Ameliorate Liver Fibrosis in Rats: Modulation of cAMP\/CREB\/TLR4 Inflammatory and Fibrogenic Pathways","volume":"222","author":"Essam","year":"2019","journal-title":"Life Sci."},{"key":"B12","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1067\/mai.2001.119555","article-title":"Cyclic Nucleotide Phosphodiesterases","volume":"108","author":"Essayan","year":"2001","journal-title":"J. Allergy Clin. Immunol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"1291","DOI":"10.1152\/physrev.00032.2009","article-title":"Regulation of the Pulmonary Circulation in the Fetus and Newborn","volume":"90","author":"Gao","year":"2010","journal-title":"Physiol. Rev."},{"key":"B14","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1054\/bjom.1999.0062","article-title":"Clinical and Functional Staging of Oral Submucous Fibrosis","volume":"38","author":"Haider","year":"2000","journal-title":"Br. J. Oral Maxillofac. Surg."},{"key":"B15","doi-asserted-by":"publisher","first-page":"e51806","DOI":"10.1371\/journal.pone.0051806","article-title":"Activation of TGF-\u03b2 Pathway by Areca Nut Constituents: a Possible Cause of Oral Submucous Fibrosis","volume":"7","author":"Khan","year":"2012","journal-title":"PLoS One"},{"key":"B16","doi-asserted-by":"publisher","first-page":"450","DOI":"10.1111\/j.1600-0714.1995.tb01132.x","article-title":"Betel Quid Chewing, Cigarette Smoking and Alcohol Consumption Related to Oral Cancer in Taiwan","volume":"24","author":"Ko","year":"1995","journal-title":"J. Oral Pathol. Med."},{"key":"B17","doi-asserted-by":"publisher","first-page":"4751","DOI":"10.1091\/mbc.e09-01-0019","article-title":"Expression and Activity of Phosphodiesterase Isoforms during Epithelial Mesenchymal Transition: the Role of Phosphodiesterase 4","volume":"20","author":"Kolosionek","year":"2009","journal-title":"Mol. Biol. Cel"},{"key":"B18","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1016\/s0014-5793(03)00563-5","article-title":"cAMP Effector Mechanisms. Novel Twists for an 'old' Signaling System","volume":"546","author":"Kopperud","year":"2003","journal-title":"FEBS Lett."},{"key":"B19","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1186\/s13046-018-0950-9","article-title":"LncRNA XIST\/miR-34a axis Modulates the Cell Proliferation and Tumor Growth of Thyroid Cancer through MET-Pi3k-AKT Signaling","volume":"37","author":"Liu","year":"2018","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"B20","doi-asserted-by":"publisher","first-page":"21146","DOI":"10.1038\/srep21146","article-title":"BATMAN-TCM: a Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine","volume":"6","author":"Liu","year":"2016","journal-title":"Sci. Rep."},{"key":"B21","doi-asserted-by":"publisher","first-page":"3583","DOI":"10.1002\/jcp.27039","article-title":"RELA\/NEAT1\/miR-302a-3p\/RELA Feedback Loop Modulates Pancreatic Ductal Adenocarcinoma Cell Proliferation and Migration","volume":"234","author":"Luo","year":"2019","journal-title":"J. Cel Physiol"},{"key":"B22","doi-asserted-by":"publisher","first-page":"1133","DOI":"10.1136\/annrheumdis-2016-210189","article-title":"Inhibition of Phosphodiesterase 4 (PDE4) Reduces Dermal Fibrosis by Interfering with the Release of Interleukin-6 from M2 Macrophages","volume":"76","author":"Maier","year":"2017","journal-title":"Ann. Rheum. Dis."},{"key":"B23","doi-asserted-by":"publisher","first-page":"220","DOI":"10.1016\/s0140-6736(05)62388-4","article-title":"Oral Submucous Fibrosis and Copper","volume":"350","author":"Meghji","year":"1997","journal-title":"Lancet"},{"key":"B24","doi-asserted-by":"publisher","first-page":"21740","DOI":"10.1073\/pnas.2007910117","article-title":"Simple Binding of Protein Kinase A Prior to Phosphorylation Allows CFTR Anion Channels to Be Opened by Nucleotides","volume":"117","author":"Mih\u00e1lyi","year":"2020","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B25","doi-asserted-by":"publisher","first-page":"552","DOI":"10.1016\/j.jobcr.2020.08.004","article-title":"Medicinal Management of Oral Submucous Fibrosis in the Past Decade- A Systematic Review","volume":"10","author":"More","year":"2020","journal-title":"J. Oral Biol. Craniofac. Res."},{"key":"B26","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1016\/j.jobcr.2019.06.016","article-title":"Proposed Clinical Definition for Oral Submucous Fibrosis","volume":"9","author":"More","year":"2019","journal-title":"J. Oral Biol. Craniofac. Res."},{"key":"B27","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1002\/path.2786","article-title":"Betel-derived Alkaloid Up-Regulates Keratinocyte Alphavbeta6 Integrin Expression and Promotes Oral Submucous Fibrosis","volume":"223","author":"Moutasim","year":"2011","journal-title":"J. Pathol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"557","DOI":"10.1016\/j.oraloncology.2006.06.007","article-title":"Elevated Expression of NF-kappaB in Oral Submucous Fibrosis-Eevidence for NF-kappaB Induction by Safrole in Human Buccal Mucosal Fibroblasts","volume":"43","author":"Ni","year":"2007","journal-title":"Oral Oncol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.devcel.2006.05.011","article-title":"DeltaEF1 Mediates TGF-Beta Signaling in Vascular Smooth Muscle Cell Differentiation","volume":"11","author":"Nishimura","year":"2006","journal-title":"Dev. Cel"},{"key":"B30","doi-asserted-by":"publisher","first-page":"112045","DOI":"10.1016\/j.jep.2019.112045","article-title":"Anti-osteoporosis Effects of Osteoking via Reducing Reactive Oxygen Species","volume":"244","author":"Qin","year":"2019","journal-title":"J. Ethnopharmacol"},{"key":"B31","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/s40463-020-0399-7","article-title":"Oral Submucous Fibrosis: a Contemporary Narrative Review with a Proposed Inter-professional Approach for an Early Diagnosis and Clinical Management","volume":"49","author":"Rao","year":"2020","journal-title":"J. Otolaryngol. Head Neck Surg."},{"key":"B32","doi-asserted-by":"publisher","first-page":"e47","DOI":"10.1093\/nar\/gkv007","article-title":"Limma powers Differential Expression Analyses for RNA-Sequencing and Microarray Studies","volume":"43","author":"Ritchie","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"B33","doi-asserted-by":"publisher","first-page":"728","DOI":"10.1016\/s1368-8375(03)00101-5","article-title":"Augmented mRNA Expression of Tissue Inhibitor of Metalloproteinase-1 in Buccal Mucosal Fibroblasts by Arecoline and Safrole as a Possible Pathogenesis for Oral Submucous Fibrosis","volume":"39","author":"Shieh","year":"2003","journal-title":"Oral Oncol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"670","DOI":"10.1177\/0271678X17749392","article-title":"Tonic Regulation of Middle Meningeal Artery Diameter by ATP-Sensitive Potassium Channels","volume":"39","author":"Syed","year":"2019","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"B35","doi-asserted-by":"publisher","first-page":"D447","DOI":"10.1093\/nar\/gku1003","article-title":"STRING V10: Protein-Protein Interaction Networks, Integrated over the Tree of Life","volume":"43","author":"Szklarczyk","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"B36","doi-asserted-by":"publisher","first-page":"561","DOI":"10.1016\/j.oraloncology.2005.08.005","article-title":"Oral Submucous Fibrosis: Review on Aetiology and Pathogenesis","volume":"42","author":"Tilakaratne","year":"2006","journal-title":"Oral Oncol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"940","DOI":"10.1016\/j.oraloncology.2005.05.006","article-title":"The Upregulation of Insulin-like Growth Factor-1 in Oral Submucous Fibrosis","volume":"41","author":"Tsai","year":"2005","journal-title":"Oral Oncol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"2104","DOI":"10.1111\/j.1743-6109.2010.01808.x","article-title":"Expression of Cyclic AMP-dependent Protein Kinase Isoforms in Human Cavernous Arteries: Functional Significance and Relation to Phosphodiesterase Type 4","volume":"7","author":"Waldkirch","year":"2010","journal-title":"J. Sex. Med."},{"key":"B39","doi-asserted-by":"publisher","first-page":"682","DOI":"10.1038\/s41419-020-02851-w","article-title":"The circEPSTI1\/mir-942-5p\/LTBP2 axis Regulates the Progression of OSCC in the Background of OSF via EMT and the PI3K\/Akt\/mTOR Pathway","volume":"11","author":"Wang","year":"2020","journal-title":"Cell Death Dis"},{"key":"B40","doi-asserted-by":"publisher","first-page":"4008","DOI":"10.3390\/ijms21114008","article-title":"A Novel, Pan-PDE Inhibitor Exerts Anti-fibrotic Effects in Human Lung Fibroblasts via Inhibition of TGF-\u03b2 Signaling and Activation of cAMP\/PKA Signaling","volume":"21","author":"W\u00f3jcik-Pszczo\u0142a","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B41","doi-asserted-by":"publisher","first-page":"193","DOI":"10.2147\/CCID.S80576","article-title":"Oral Submucous Fibrosis: an Update","volume":"8","author":"Wollina","year":"2015","journal-title":"Clin. Cosmet. Investig. Dermatol."},{"key":"B42","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1016\/s1368-8375(02)00123-9","article-title":"The Upregulation of Type I Plasminogen Activator Inhibitor in Oral Submucous Fibrosis","volume":"39","author":"Yang","year":"2003","journal-title":"Oral Oncol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"6386","DOI":"10.1073\/pnas.0801490105","article-title":"The Cyclic AMP Effector Epac Integrates Pro- and Anti-fibrotic Signals","volume":"105","author":"Yokoyama","year":"2008","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B44","doi-asserted-by":"publisher","first-page":"e55122","DOI":"10.1371\/journal.pone.0055122","article-title":"Elevation of S100A4 Expression in Buccal Mucosal Fibroblasts by Arecoline: Involvement in the Pathogenesis of Oral Submucous Fibrosis","volume":"8","author":"Yu","year":"2013","journal-title":"PLoS One"},{"key":"B45","doi-asserted-by":"publisher","first-page":"3425","DOI":"10.1158\/0008-5472.CAN-12-3840","article-title":"miR145 Targets the SOX9\/ADAM17 axis to Inhibit Tumor-Initiating Cells and IL-6-mediated Paracrine Effects in Head and Neck Cancer","volume":"73","author":"Yu","year":"2013","journal-title":"Cancer Res."},{"key":"B46","doi-asserted-by":"publisher","first-page":"37004","DOI":"10.1038\/srep37004","article-title":"Aberrant SSEA-4 Upregulation Mediates Myofibroblast Activity to Promote Pre-cancerous Oral Submucous Fibrosis","volume":"6","author":"Yu","year":"2016","journal-title":"Sci. Rep."},{"key":"B47","doi-asserted-by":"publisher","first-page":"1688","DOI":"10.2174\/138161209788168092","article-title":"Cyclic AMP-specific Phosphodiesterase-4 as a Target for the Development of Antidepressant Drugs","volume":"15","author":"Zhang","year":"2009","journal-title":"Curr. Pharm. Des."},{"key":"B48","doi-asserted-by":"publisher","first-page":"681","DOI":"10.1002\/ijc.11114","article-title":"Independent and Combined Effects of Tobacco Smoking, Chewing and Alcohol Drinking on the Risk of Oral, Pharyngeal and Esophageal Cancers in Indian Men","volume":"105","author":"Znaor","year":"2003","journal-title":"Int. J. Cancer"}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2021.722040\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,8]],"date-time":"2021-11-08T05:36:09Z","timestamp":1636349769000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2021.722040\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,8]]},"references-count":48,"alternative-id":["10.3389\/fphar.2021.722040"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2021.722040","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,8]]},"article-number":"722040"}}