{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T01:08:38Z","timestamp":1770685718284,"version":"3.49.0"},"reference-count":39,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2019,1,18]],"date-time":"2019-01-18T00:00:00Z","timestamp":1547769600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["NRF-2017M3A9E8033225"],"award-info":[{"award-number":["NRF-2017M3A9E8033225"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["NRF 2017R1C1B5017879"],"award-info":[{"award-number":["NRF 2017R1C1B5017879"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase [NOX] enzymes serve several hemostatic and host defense functions in various lung diseases, but the role of NOX4 signaling in tuberculous pleurisy is not well understood. The role of NOX4 signaling in tuberculous pleural fibrosis was studied using invitro pleural mesothelial cell (PMC) experiments and a murine model of Mycobacterium bovis bacillus Calmette\u2013Gu\u00e9rin (BCG) pleural infection. The production of NOX4 reactive oxygen species (NOX4\u2013ROS) and the epithelial mesenchymal transition (EMT) in PMCs were both induced by heat-killed mycobacterium tuberculosis (HKMT). In cultured PMCs, HKMT-induced collagen-1 synthesis and EMT were blocked by pretreatment with small interfering RNA (siRNA) NOX4. Moreover, NOX4\u2013ROS production and subsequent fibrosis were reduced by treatment with losartan and the toll-like receptor 4 (TLR4) inhibitor TAK-242. The HKMT-induced EMT and intracellular ROS production were mediated by NOX4 via the activation of extracellular signal-regulated kinase (ERK) signaling. Finally, in a BCG-induced pleurisy model, recruitment of inflammatory pleural cells, release of inflammatory cytokines, and thickened mesothelial fibrosis were attenuated by SiNOX4 compared to SiCon. Our study identified that HKMT-induced pleural fibrosis is mediated by NOX4\u2013ERK\u2013ROS via TLR4 and Angiotensin II receptor type1 (AT1R). There results suggest that NOX4 may be a novel therapeutic target for intervention in tuberculous pleural fibrosis.<\/jats:p>","DOI":"10.3390\/jcm8010116","type":"journal-article","created":{"date-parts":[[2019,1,18]],"date-time":"2019-01-18T05:41:08Z","timestamp":1547790068000},"page":"116","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Inhibition of NADPH Oxidase 4 (NOX4) Signaling Attenuates Tuberculous Pleural Fibrosis"],"prefix":"10.3390","volume":"8","author":[{"given":"Youngmi","family":"Kim","sequence":"first","affiliation":[{"name":"Institute of New frontier Research, Hallym University College of Medicine, Chuncheon 24253, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"So Yeong","family":"Park","sequence":"additional","affiliation":[{"name":"Institute of New frontier Research, Hallym University College of Medicine, Chuncheon 24253, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Harry","family":"Jung","sequence":"additional","affiliation":[{"name":"Institute of New frontier Research, Hallym University College of Medicine, Chuncheon 24253, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"You Sun","family":"Noh","sequence":"additional","affiliation":[{"name":"Institute of New frontier Research, Hallym University College of Medicine, Chuncheon 24253, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jae Jun","family":"Lee","sequence":"additional","affiliation":[{"name":"Institute of New frontier Research, Hallym University College of Medicine, Chuncheon 24253, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3132-7706","authenticated-orcid":false,"given":"Ji Young","family":"Hong","sequence":"additional","affiliation":[{"name":"Institute of New frontier Research, Hallym University College of Medicine, Chuncheon 24253, Korea"},{"name":"Division of Pulmonary, Allergy and Critical Care Medicine, Department of Internal Medicine, Chuncheon Sacred Heart Hospital, Hallym University Medical Center, Chuncheon 24235, Korea"},{"name":"Lung Research Institute of Hallym University College of Medicine, Chuncheon 24253, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,1,18]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2017). Global Tuberculosis Report 2017, World Health Organization."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1378\/chest.99.4.883","article-title":"Tuberculous pleural effusion. Twenty-year experience","volume":"99","author":"Seibert","year":"1991","journal-title":"Chest"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1378\/chest.112.5.1293","article-title":"Are pleural fluid parameters related to the development of residual pleural thickening in tuberculosis?","volume":"112","author":"Villena","year":"1997","journal-title":"Chest"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"L145","DOI":"10.1152\/ajplung.00348.2015","article-title":"Activation of calpain by renin-angiotensin system in pleural mesothelial cells mediates tuberculous pleural fibrosis","volume":"311","author":"Yang","year":"2016","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bocchino, M., Agnese, S., Fagone, E., Svegliati, S., Grieco, D., Vancheri, C., Gabrielli, A., Sanduzzi, A., and Avvedimento, E.V. (2010). Reactive oxygen species are required for maintenance and differentiation of primary lung fibroblasts in idiopathic pulmonary fibrosis. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0014003"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.1038\/nm.2005","article-title":"NADPH oxidase-4 mediates myofibroblast activation and fibrogenic responses to lung injury","volume":"15","author":"Hecker","year":"2009","journal-title":"Nat. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1086","DOI":"10.1016\/j.biocel.2011.04.003","article-title":"Signalling pathways from NADPH oxidase-4 to idiopathic pulmonary fibrosis","volume":"43","author":"Crestani","year":"2011","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1136\/thx.2009.113456","article-title":"NOX4\/NADPH oxidase expression is increased in pulmonary fibroblasts from patients with idiopathic pulmonary fibrosis and mediates TGFbeta1-induced fibroblast differentiation into myofibroblasts","volume":"65","author":"Amara","year":"2010","journal-title":"Thorax"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4392","DOI":"10.18632\/oncotarget.2025","article-title":"NOX4 promotes non-small cell lung cancer cell proliferation and metastasis through positive feedback regulation of PI3K\/Akt signaling","volume":"5","author":"Zhang","year":"2014","journal-title":"Oncotarget"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1164\/rccm.201107-1281OC","article-title":"Increased nicotinamide adenine dinucleotide phosphate oxidase 4 expression mediates intrinsic airway smooth muscle hypercontractility in asthma","volume":"185","author":"Sutcliffe","year":"2012","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1089\/ars.2008.2203","article-title":"Role of Nox4 and Nox2 in hyperoxia-induced reactive oxygen species generation and migration of human lung endothelial cells","volume":"11","author":"Pendyala","year":"2009","journal-title":"Antioxid. Redox Signal."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.tiv.2012.09.002","article-title":"Comparative cytological responses of lung epithelial and pleural mesothelial cells following in vitro exposure to nanoscale SiO2","volume":"27","author":"Berg","year":"2013","journal-title":"Toxicol. In Vitro"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.cyto.2017.02.021","article-title":"TLR2 contributes to trigger immune response of pleural mesothelial cells against Mycobacterium bovis BCG and M. tuberculosis infection","volume":"95","author":"Hwanga","year":"2017","journal-title":"Cytokine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2364","DOI":"10.1016\/j.ajpath.2016.05.015","article-title":"Tumor Necrosis Factor and Its Receptors Are Crucial to Control Mycobacterium bovis Bacillus Calmette-Guerin Pleural Infection in a Murine Model","volume":"186","author":"Vesin","year":"2016","journal-title":"Am. J. Pathol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Segueni, N., Vigne, S., Palmer, G., Bourigault, M.L., Olleros, M.L., Vesin, D., Garcia, I., Ryffel, B., Quesniaux, V.F., and Gabay, C. (2015). Limited Contribution of IL-36 versus IL-1 and TNF Pathways in Host Response to Mycobacterial Infection. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0126058"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1136\/jcp.41.4.467","article-title":"Simple method of estimating severity of pulmonary fibrosis on a numerical scale","volume":"41","author":"Ashcroft","year":"1988","journal-title":"J. Clin. Pathol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"331","DOI":"10.2353\/ajpath.2006.050435","article-title":"EphrinA1 inhibits vascular endothelial growth factor-induced intracellular signaling and suppresses retinal neovascularization and blood-retinal barrier breakdown","volume":"168","author":"Ojima","year":"2006","journal-title":"Am. J. Pathol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"507","DOI":"10.7326\/0003-4819-77-4-507","article-title":"Pleural effusions: The diagnostic separation of transudates and exudates","volume":"77","author":"Light","year":"1972","journal-title":"Ann. Intern. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"12126","DOI":"10.1074\/jbc.M113.521245","article-title":"Histone deacetylase-3 mediates positive feedback relationship between anaphylaxis and tumor metastasis","volume":"289","author":"Eom","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"51","DOI":"10.3349\/ymj.2011.52.1.51","article-title":"Epithelial to mesenchymal transition of mesothelial cells in tuberculous pleurisy","volume":"52","author":"Kim","year":"2011","journal-title":"Yonsei Med. J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"916310","DOI":"10.4061\/2011\/916310","article-title":"Oxidative Stress and Vascular Damage in Hypertension: Role of Angiotensin II","volume":"2011","author":"Virdis","year":"2011","journal-title":"Int. J. Hypertens."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"De Batista, P.R., Palacios, R., Martin, A., Hernanz, R., Medici, C.T., Silva, M.A., Rossi, E.M., Aguado, A., Vassallo, D.V., and Salaices, M. (2014). Toll-like receptor 4 upregulation by angiotensin II contributes to hypertension and vascular dysfunction through reactive oxygen species production. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0104020"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1038\/hr.2015.55","article-title":"TLR4 is a critical regulator of angiotensin II-induced vascular remodeling: The roles of extracellular SOD and NADPH oxidase","volume":"38","author":"Nakashima","year":"2015","journal-title":"Hypertens. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2838","DOI":"10.1089\/ars.2013.5608","article-title":"NADPH oxidases in lung health and disease","volume":"20","author":"Bernard","year":"2014","journal-title":"Antioxid. Redox Signal."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1016\/j.freeradbiomed.2009.07.023","article-title":"Nox proteins in signal transduction","volume":"47","author":"Brown","year":"2009","journal-title":"Free Radic. Biol. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1007\/s00281-008-0120-9","article-title":"NOX enzymes and Toll-like receptor signaling","volume":"30","author":"Mkaddem","year":"2008","journal-title":"Semin. Immunopathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1165\/rcmb.2012-0242OC","article-title":"Role of nicotinamide adenine dinucleotide phosphate-reduced oxidase proteins in Pseudomonas aeruginosa-induced lung inflammation and permeability","volume":"48","author":"Fu","year":"2013","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1007\/s00408-013-9469-1","article-title":"Angiotensin-converting enzyme (ACE) gene polymorphisms are associated with idiopathic pulmonary fibrosis","volume":"191","author":"Uh","year":"2013","journal-title":"Lung"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1766","DOI":"10.1097\/HJH.0b013e328356766f","article-title":"Nebivolol reduces cardiac angiotensin II, associated oxidative stress and fibrosis but not arterial pressure in salt-loaded spontaneously hypertensive rats","volume":"30","author":"Varagic","year":"2012","journal-title":"J. Hypertens."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4079","DOI":"10.1093\/ndt\/gfr217","article-title":"Human peritoneal mesothelial cells respond to bacterial ligands through a specific subset of Toll-like receptors","volume":"26","author":"Colmont","year":"2011","journal-title":"Nephrol. Dial. Transplant."},{"key":"ref_31","first-page":"1289","article-title":"Endotoxin-induced chemokine expression in murine peritoneal mesothelial cells: The role of toll-like receptor 4","volume":"15","author":"Kato","year":"2004","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1007\/s10753-016-0464-6","article-title":"Angiotensin II-Induced Early and Late Inflammatory Responses Through NOXs and MAPK Pathways","volume":"40","author":"Zhang","year":"2017","journal-title":"Inflammation"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"E132","DOI":"10.1152\/ajpendo.00629.2010","article-title":"Angiotensin II plays a critical role in diabetic pulmonary fibrosis most likely via activation of NADPH oxidase-mediated nitrosative damage","volume":"301","author":"Yang","year":"2011","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1089\/ars.2013.5818","article-title":"The angiotensin-converting enzyme 2\/angiotensin (1-7) \/Mas axis protects against lung fibroblast migration and lung fibrosis by inhibiting the NOX4-derived ROS-mediated RhoA\/Rho kinase pathway","volume":"22","author":"Meng","year":"2015","journal-title":"Antioxid. Redox Signal."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1152\/physrev.00044.2005","article-title":"The NOX family of ROS-generating NADPH oxidases: Physiology and pathophysiology","volume":"87","author":"Bedard","year":"2007","journal-title":"Physiol. Rev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"L433","DOI":"10.1152\/ajplung.00128.2016","article-title":"Differential regulation of autophagy and mitophagy in pulmonary diseases","volume":"311","author":"Aggarwal","year":"2016","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"L218","DOI":"10.1152\/ajplung.00162.2017","article-title":"Role of apoptosis and autophagy in tuberculosis","volume":"313","author":"Lam","year":"2017","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.yexcr.2017.03.039","article-title":"MiR-23a-5p modulates mycobacterial survival and autophagy during mycobacterium tuberculosis infection through TLR2\/MyD88\/NF-kappaB pathway by targeting TLR2","volume":"354","author":"Gu","year":"2017","journal-title":"Exp. Cell Res."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Pan, M., Zheng, Z., Chen, Y., Sun, N., Zheng, B., Yang, Q., Zhang, Y., Li, X., and Meng, Y. (2018). Angiotensin- (1-7) Attenuated Cigarette Smoking-related Pulmonary Fibrosis via Improving the Impaired Autophagy Caused by NOX4-dependent ROS. Am. J. Respir. Cell Mol. Biol.","DOI":"10.1165\/rcmb.2017-0284OC"}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/8\/1\/116\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:27:05Z","timestamp":1760185625000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/8\/1\/116"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,1,18]]},"references-count":39,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,1]]}},"alternative-id":["jcm8010116"],"URL":"https:\/\/doi.org\/10.3390\/jcm8010116","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,1,18]]}}}