{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:46:42Z","timestamp":1785422802738,"version":"3.56.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T00:00:00Z","timestamp":1770163200000},"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. Med."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Chronic ethanol exposure increases susceptibility to fibroproliferative maladaptive repair following acute lung injury. Ethanol disrupts molecular circadian rhythms in multiple organs, contributing to liver steatosis and renal fibrosis. Because circadian disruption is linked to TGF\u03b2 activation and tissue fibrosis, we hypothesized that ethanol alters lung circadian signaling and promotes profibrotic responses in lung fibroblasts through modulation of TGF\u03b2 and \u03b1-SMA expression.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Lung slices from PER2-luciferase reporter mice fed with 20% (v\/v) ethanol in drinking water for 8\u202fweeks or only water (control) for 8\u202fweeks were analyzed for real-time bioluminescent PER2 rhythms over 7\u202fdays. Lungs from control and ethanol-fed C57BL\/6J mice were collected every 4\u202fh over 24\u202fh to assess rhythmicity of selected core clock genes and selected profibrotic markers mRNA expression. Primary murine lung fibroblasts (PLF) were treated with ethanol and evaluated for circadian gene and protein expression. ROR\u03b1 function was interrogated using siRNA knockdown and pharmacological agonist\/inverse agonist, followed by analysis of TGF\u03b2, \u03b1-SMA, and fibronectin protein levels.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Chronic ethanol ingestion lengthened the circadian period by ~2\u202fh (\n                      <jats:italic>p<\/jats:italic>\n                      &amp;lt;\u202f0.05) and induced a ~7% phase shift in PER2 rhythms in lung slices. Ethanol altered oscillatory patterns of core clock genes (\n                      <jats:italic>Bmal1, Clock, Ror\u03b1, Rev-erb\u03b1<\/jats:italic>\n                      ) and profibrotic markers (\n                      <jats:italic>Tgf\u03b2, \u03b1-SMA, Fn1<\/jats:italic>\n                      ) in mouse lungs.\n                      <jats:italic>In vitro<\/jats:italic>\n                      , ethanol suppressed BMAL1 and ROR\u03b1 expression in PLF. Activation of ROR\u03b1 with agonist SR1078 reversed ethanol-induced TGF\u03b2 and \u03b1-SMA upregulation, whereas ROR\u03b1 reverse agonist (SR3335) mimicked ethanol\u2019s effects. Lastly, the silencing of ROR\u03b1 gene expression significantly induced TGF\u03b2 and \u03b1-SMA, with a trend toward an increase in Fn1.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Ethanol disrupts circadian signaling and enhances profibrotic gene expression in lung fibroblasts, partly through suppression of ROR\u03b1. ROR\u03b1 activation mitigates these effects, identifying ROR\u03b1 as a potential therapeutic target for ethanol-related maladaptive lung repair.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmed.2026.1719787","type":"journal-article","created":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T06:47:06Z","timestamp":1770187626000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Downregulation of ROR\u03b1 by alcohol promotes TGF\u03b2 and \u03b1-SMA expression in mouse lung fibroblasts"],"prefix":"10.3389","volume":"13","author":[{"given":"Xian","family":"Fan","sequence":"first","affiliation":[{"name":"Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Department of Medicine, Emory University School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Tao","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Department of Medicine, Emory University School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bum-Yong","family":"Kang","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Allergy, Sleep, and Cystic Fibrosis, Department of Pediatrics, Emory University School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Diaz","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, Morehouse School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenkichi","family":"Baba","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, Morehouse School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gianluca","family":"Tosini","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, Morehouse School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin","family":"Guo","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Department of Medicine, Emory University School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samantha M.","family":"Yeligar","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Department of Medicine, Emory University School of Medicine","place":["Atlanta, GA, United States"]},{"name":"Atlanta Veterans Affairs Health Care System","place":["Decatur, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Viranuj","family":"Sueblinvong","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Department of Medicine, Emory University School of Medicine","place":["Atlanta, GA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,2,4]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1038\/s41392-022-00899-y","article-title":"Molecular regulations of circadian rhythm and implications for physiology and diseases","volume":"7","author":"Fagiani","year":"2022","journal-title":"Signal Transduct Target Ther"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1146\/annurev-neuro-060909-153128","article-title":"Central and peripheral circadian clocks in mammals","volume":"35","author":"Mohawk","year":"2012","journal-title":"Annu Rev Neurosci"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"4753","DOI":"10.1038\/ncomms5753","article-title":"Circadian rhythm reprogramming during lung inflammation","volume":"5","author":"Haspel","year":"2014","journal-title":"Nat Commun"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1016\/j.bbrc.2023.05.092","article-title":"Rev-erbalpha agonists suppresses TGFbeta1-induced fibroblast-to-myofibroblast transition and pro-fibrotic phenotype in human lung fibroblasts","volume":"669","author":"Prasad","year":"2023","journal-title":"Biochem Biophys Res Commun"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"1139","DOI":"10.1073\/pnas.1912109117","article-title":"The circadian clock protein REVERBalpha inhibits pulmonary fibrosis development","volume":"117","author":"Cunningham","year":"2020","journal-title":"Proc Natl Acad Sci USA"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"106110","DOI":"10.1016\/j.biocel.2021.106110","article-title":"Circadian clock dysfunction of epithelial cells in pulmonary diseases","volume":"141","author":"Gao","year":"2021","journal-title":"Int J Biochem Cell Biol"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1056\/NEJMra1802337","article-title":"Circadian mechanisms in medicine","volume":"384","author":"Allada","year":"2021","journal-title":"N Engl J Med"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"668","DOI":"10.1111\/acer.13612","article-title":"Circadian mechanisms in alcohol use disorder and tissue injury","volume":"42","author":"Davis","year":"2018","journal-title":"Alcohol Clin Exp Res"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"2504","DOI":"10.3390\/biom5042504","article-title":"The molecular circadian clock and alcohol-induced liver injury","volume":"5","author":"Udoh","year":"2015","journal-title":"Biomolecules"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"869","DOI":"10.1097\/01.CCM.0000055389.64497.11","article-title":"Chronic alcohol abuse is associated with an increased incidence of acute respiratory distress syndrome and severity of multiple organ dysfunction in patients with septic shock","volume":"31","author":"Moss","year":"2003","journal-title":"Crit Care Med"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"1839","DOI":"10.1111\/acer.15174","article-title":"Ethanol-exposed lung fibroblasts cause airway epithelial barrier dysfunction","volume":"47","author":"Sueblinvong","year":"2023","journal-title":"Alcohol Clin Exp Res"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"1952","DOI":"10.1111\/j.1530-0277.2012.01825.x","article-title":"Ethanol (EtOH)-induced TGF-beta1 and reactive oxygen species production are necessary for EtOH-induced alveolar macrophage dysfunction and induction of alternative activation","volume":"36","author":"Brown","year":"2012","journal-title":"Alcohol Clin Exp Res"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"L363","DOI":"10.1152\/ajplung.00141.2005","article-title":"Angiotensin II mediates glutathione depletion, transforming growth factor-beta1 expression, and epithelial barrier dysfunction in the alcoholic rat lung","volume":"289","author":"Bechara","year":"2005","journal-title":"Am J Physiol Lung Cell Mol Physiol"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"L1212","DOI":"10.1152\/ajplung.00042.2014","article-title":"The relative balance of GM-CSF and TGF-beta1 regulates lung epithelial barrier function","volume":"308","author":"Overgaard","year":"2015","journal-title":"Am J Physiol Lung Cell Mol Physiol"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1016\/j.alcohol.2007.04.010","article-title":"Chronic alcohol ingestion alters claudin expression in the alveolar epithelium of rats","volume":"41","author":"Fernandez","year":"2007","journal-title":"Alcohol"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"1866","DOI":"10.1111\/j.1530-0277.2011.01531.x","article-title":"Chronic alcohol ingestion exacerbates lung epithelial barrier dysfunction in HIV-1 transgenic rats","volume":"35","author":"Fan","year":"2011","journal-title":"Alcohol Clin Exp Res"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1165\/rcmb.2012-0334oc","article-title":"Alcohol causes alveolar epithelial oxidative stress by inhibiting the nuclear factor (erythroid-derived 2)-like 2-antioxidant response element signaling pathway","volume":"48","author":"Jensen","year":"2013","journal-title":"Am J Respir Cell Mol Biol"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"4338","DOI":"10.1182\/blood-2009-04-214817","article-title":"microRNA miR-144 modulates oxidative stress tolerance and associates with anemia severity in sickle cell disease","volume":"116","author":"Sangokoya","year":"2010","journal-title":"Blood"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"e003","DOI":"10.1621\/nrs.07003","article-title":"Retinoid-related orphan receptors (RORs): critical roles in development, immunity, circadian rhythm, and cellular metabolism","volume":"7","author":"Jetten","year":"2009","journal-title":"Nucl Recept Signal"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"13959","DOI":"10.1074\/jbc.M117.782391","article-title":"Retinoic acid receptor-related orphan receptor alpha stimulates adipose tissue inflammation by modulating endoplasmic reticulum stress","volume":"292","author":"Liu","year":"2017","journal-title":"J Biol Chem"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"1278","DOI":"10.1038\/s12276-021-00679-8","article-title":"Unraveling the physiological roles of retinoic acid receptor-related orphan receptor alpha","volume":"53","author":"Lee","year":"2021","journal-title":"Exp Mol Med"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"102059","DOI":"10.1016\/j.jbc.2022.102059","article-title":"Retinoid orphan nuclear receptor alpha (RORalpha) suppresses the epithelial-mesenchymal transition (EMT) by directly repressing snail transcription","volume":"298","author":"Xiong","year":"2022","journal-title":"J Biol Chem"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"L975","DOI":"10.1152\/ajplung.00003.2004","article-title":"Ethanol stimulates the expression of fibronectin in lung fibroblasts via kinase-dependent signals that activate CREB","volume":"288","author":"Roman","year":"2005","journal-title":"Am J Physiol Lung Cell Mol Physiol"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"336","DOI":"10.1111\/acer.12232","article-title":"Chronic alcohol ingestion primes the lung for bleomycin-induced fibrosis in mice","volume":"38","author":"Sueblinvong","year":"2014","journal-title":"Alcohol Clin Exp Res"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1165\/rcmb.2017-0289oc","article-title":"Quercetin enhances ligand-induced apoptosis in senescent idiopathic pulmonary fibrosis fibroblasts and reduces lung fibrosis in vivo","volume":"60","author":"Hohmann","year":"2019","journal-title":"Am J Respir Cell Mol Biol"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"3654","DOI":"10.1038\/s41598-023-30844-0","article-title":"Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts","volume":"13","author":"Badaro-Garcia","year":"2023","journal-title":"Sci Rep"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"137","DOI":"10.4137\/BCBCR.S9673","article-title":"Oscillation of clock and clock controlled genes induced by serum shock in human breast epithelial and breast cancer cells: regulation by melatonin","volume":"6","author":"Xiang","year":"2012","journal-title":"Breast Cancer"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"1168","DOI":"10.3748\/wjg.15.1168","article-title":"In vitro and in vivo models of acute alcohol exposure","volume":"15","author":"Dolganiuc","year":"2009","journal-title":"World J Gastroenterol"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"32295","DOI":"10.1038\/s41598-025-15040-6","article-title":"Circular RNA RORbeta regulates TGFbetaR1 in alcohol-induced fibroblast-to-myofibroblast differentiation","volume":"15","author":"Sueblinvong","year":"2025","journal-title":"Sci Rep"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"2686","DOI":"10.1111\/j.1460-9568.2008.06231.x","article-title":"The circadian pacemaker in the cultured suprachiasmatic nucleus from pup mice is highly sensitive to external perturbation","volume":"27","author":"Nishide","year":"2008","journal-title":"Eur J Neurosci"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1097\/MAJ.0b013e318234c132","article-title":"Predisposition for disrepair in the aged lung","volume":"344","author":"Sueblinvong","year":"2012","journal-title":"Am J Med Sci"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"372","DOI":"10.1177\/0748730410379711","article-title":"JTK_CYCLE: an efficient nonparametric algorithm for detecting rhythmic components in genome-scale data sets","volume":"25","author":"Hughes","year":"2010","journal-title":"J Biol Rhythm"},{"key":"ref33","doi-asserted-by":"publisher","first-page":"1029","DOI":"10.1021\/cb100223d","article-title":"Identification of SR1078, a synthetic agonist for the orphan nuclear receptors RORalpha and RORgamma","volume":"5","author":"Wang","year":"2010","journal-title":"ACS Chem Biol"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"2731","DOI":"10.1111\/acer.12563","article-title":"TGFbeta1 mediates alcohol-induced Nrf2 suppression in lung fibroblasts","volume":"38","author":"Sueblinvong","year":"2014","journal-title":"Alcohol Clin Exp Res"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"eabg5174","DOI":"10.1126\/sciadv.abg5174","article-title":"Intercellular coupling between peripheral circadian oscillators by TGF-\u03b2 signaling","volume":"7","author":"Finger","year":"2021","journal-title":"Sci Adv"},{"key":"ref36","doi-asserted-by":"publisher","first-page":"e1008459","DOI":"10.1371\/journal.pcbi.1008459","article-title":"A dual-feedback loop model of the mammalian circadian clock for multi-input control of circadian phase","volume":"16","author":"Brown","year":"2020","journal-title":"PLoS Comput Biol"},{"key":"ref37","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1186\/s13073-019-0704-0","article-title":"Genomics of circadian rhythms in health and disease","volume":"11","author":"Rijo-Ferreira","year":"2019","journal-title":"Genome Med"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1177\/07487304221141464","article-title":"Chronic alcohol consumption disrupts the skeletal muscle circadian clock in female mice","volume":"38","author":"Tice","year":"2023","journal-title":"J Biol Rhythm"},{"key":"ref39","doi-asserted-by":"publisher","first-page":"E606","DOI":"10.1152\/ajpendo.00187.2021","article-title":"Binge alcohol disrupts skeletal muscle core molecular clock independent of glucocorticoids","volume":"321","author":"Tice","year":"2021","journal-title":"Am J Physiol Endocrinol Metab"},{"key":"ref40","doi-asserted-by":"publisher","first-page":"833","DOI":"10.1016\/j.cmet.2012.11.004","article-title":"Circadian oscillations of protein-coding and regulatory RNAs in a highly dynamic mammalian liver epigenome","volume":"16","author":"Vollmers","year":"2012","journal-title":"Cell Metab"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"1295","DOI":"10.1038\/s41467-023-36896-0","article-title":"Circadian clock molecule REV-ERBalpha regulates lung fibrotic progression through collagen stabilization","volume":"14","author":"Wang","year":"2023","journal-title":"Nat Commun"}],"container-title":["Frontiers in Medicine"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmed.2026.1719787\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T09:31:22Z","timestamp":1771925482000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmed.2026.1719787\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,4]]},"references-count":41,"alternative-id":["10.3389\/fmed.2026.1719787"],"URL":"https:\/\/doi.org\/10.3389\/fmed.2026.1719787","relation":{},"ISSN":["2296-858X"],"issn-type":[{"value":"2296-858X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,4]]},"article-number":"1719787"}}