{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T20:12:32Z","timestamp":1778271152654,"version":"3.51.4"},"reference-count":129,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T00:00:00Z","timestamp":1777507200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Arachidonic acid (AA) is an emerging regulator of fibroblast activity in pulmonary fibrosis; however, the underlying intracellular mechanisms remain unclear. This study investigated the effects of AA on the free intracellular calcium concentration ([Ca2+]i), nitric oxide (NO), and reactive oxygen species (ROS) in human WI-38 lung fibroblasts. Using fluorescent imaging and pharmacological tools, we demonstrate that AA evokes a robust, concentration-dependent increase in [Ca2+]i. This response is initiated by G protein-coupled receptor 40 (GPR40), which leads to endoplasmic reticulum Ca2+ release through inositol 1,4,5-trisphosphate receptors (IP3Rs) and lysosomal Ca2+ mobilisation through two-pore channels (TPCs). Sustained Ca2+ elevation is primarily mediated by influx through transient receptor potential vanilloid 4 (TRPV4) channels, with a minor contribution from store-operated Ca2+ entry. The AA-induced Ca2+ signal stimulates endothelial NO synthase-dependent NO production, which in turn triggers ROS generation, revealing a tightly coupled Ca2+\u2013NO\u2013ROS signalling network. Our findings identify AA as a potent modulator of Ca2+ and redox signalling in lung fibroblasts, and highlight GPR40, TRPV4, IP3Rs and lysosomal TPCs as potential therapeutic targets for intervening in pulmonary fibrosis.<\/jats:p>","DOI":"10.3390\/ijms27094016","type":"journal-article","created":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T10:47:28Z","timestamp":1777546048000},"page":"4016","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Ca2+\u2013NO\u2013ROS Crosstalk Induced by Arachidonic Acid in Human Lung Fibroblasts: Implications for Pulmonary Fibrosis"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-7097-2456","authenticated-orcid":false,"given":"Karen","family":"Esmeralda S\u00e1nchez-Pluma","sequence":"first","affiliation":[{"name":"Department of Biomedicine, School of Medicine, Benem\u00e9rita Universidad Aut\u00f3noma de Puebla, Puebla 72410, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9734-315X","authenticated-orcid":false,"given":"Edgar","family":"Mart\u00ednez-Romero","sequence":"additional","affiliation":[{"name":"Department of Biomedicine, School of Medicine, Benem\u00e9rita Universidad Aut\u00f3noma de Puebla, Puebla 72410, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9685-4710","authenticated-orcid":false,"given":"Jos\u00e9 Everardo","family":"Avelino-Cruz","sequence":"additional","affiliation":[{"name":"Laboratorio de Cardiolog\u00eda Molecular, Instituto de Fisiolog\u00eda, Benem\u00e9rita Universidad Aut\u00f3noma de Puebla, Puebla 72560, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1673-6146","authenticated-orcid":false,"given":"Giorgia","family":"Scarpellino","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology \u201cL. Spallanzani\u201d, University of Pavia, 27100 Pavia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valentina","family":"Brunetti","sequence":"additional","affiliation":[{"name":"Department of Biology and Biotechnology \u201cL. Spallanzani\u201d, University of Pavia, 27100 Pavia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monica","family":"Savio","sequence":"additional","affiliation":[{"name":"Immunology and General Pathology Unit, Department of Molecular Medicine, University of Pavia, 27100 Pavia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8219-2135","authenticated-orcid":false,"given":"Luis G.","family":"V\u00e1zquez-de-Lara Cisneros","sequence":"additional","affiliation":[{"name":"Laboratorio de Medicina Experimental, Facultad de Medicina, Benem\u00e9rita Universidad Aut\u00f3noma de Puebla, Puebla 72420, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0010-0098","authenticated-orcid":false,"given":"Francesco","family":"Moccia","sequence":"additional","affiliation":[{"name":"Department of Medicine and Health Sciences \u201cV. Tiberio\u201d, University of Molise, 86100 Campobasso, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4884-0925","authenticated-orcid":false,"given":"Roberto","family":"Berra-Romani","sequence":"additional","affiliation":[{"name":"Department of Biomedicine, School of Medicine, Benem\u00e9rita Universidad Aut\u00f3noma de Puebla, Puebla 72410, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"104166","DOI":"10.1016\/j.lpm.2023.104166","article-title":"Idiopathic Pulmonary Fibrosis","volume":"52","author":"Koudstaal","year":"2023","journal-title":"Presse Med."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Effendi, W.I., and Nagano, T. (2022). Connective Tissue Growth Factor in Idiopathic Pulmonary Fibrosis: Breaking the Bridge. Int. J. Mol. 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