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Biosci. (Landmark Ed)"],"abstract":"<jats:p>Background: Although mechanical ventilation (MV)-associated excessive stretch directly damages airway epithelial cells (AECs) and contributes to ventilator-induced lung injury (VILI), the contribution of paracrine factors from airway cells, particularly airway smooth muscle cells (ASMCs) and AECs, remains poorly understood. As stretch-sensitive cells, ASMCs and AECs may respond to excessive stretch by secreting paracrine factors such as transforming growth factor \u03b21 (TGF-\u03b21), thereby disrupting airway epithelial integrity. This study compared the sensitivity of cultured ASMCs and AECs to MV-associated excessive stretch and their ability to induce paracrine factors, particularly TGF-\u03b21, that inhibit AEC proliferation in vitro. Methods: Human ASMCs and AECs (16HBE14o-) were cultured under static, physiological stretch (5% strain), or pathological excessive stretch (13% strain) conditions for 72 h. Conditioned media were collected and used to culture 16HBE14o- cells for 48 hours. Active and total TGF-\u03b21 levels in the conditioned media were measured using an enzyme-linked immunosorbent assay. Cell viability, proliferation, migration, stress fiber formation, cell cycle distribution, and Smad2 signaling were assessed using the Cell Counting Kit-8 assay, cell counting, wound healing assay, immunofluorescence, flow cytometry, quantitative reverse transcription polymerase chain reaction, and Western blotting, respectively. Results: Conditioned media derived from ASMCs, but not 16HBE14o- cells, cultured under 13% excessive stretch inhibited 16HBE14o- cell proliferation. In addition, excessive stretch enhanced TGF-\u03b21 activation in ASMCs in an integrin \u03b1V-dependent manner. Experiments using a TGF-\u03b21-neutralizing antibody and transforming growth factor \u03b2 receptor I inhibitors demonstrated that inhibition of 16HBE14o- cell proliferation by conditioned media derived from excessively stretched ASMCs was dependent on TGF-\u03b21 activation and Smad2 signaling. Conclusion: Conditioned media derived from ASMCs, but not AECs, exposed to MV-associated excessive stretch induced G1 cell cycle arrest in AECs via TGF-\u03b21\/Smad2 signaling. These findings suggest that ASMCs are more sensitive than AECs to excessive stretch-induced TGF-\u03b21 activation and may thereby contribute to disruption of airway epithelial integrity. Targeting this paracrine pathway may warrant further investigation as a potential strategy for preventing or treating VILI.<\/jats:p>","DOI":"10.31083\/fbl55462","type":"journal-article","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T16:21:51Z","timestamp":1790698911000},"source":"Crossref","is-referenced-by-count":0,"title":["Excessive Stretch of Airway Smooth Muscle (but Not Epithelial) Cells Activates Paracrine TGF-\u03b21 to Induce Smad2-Mediated G1 Arrest in Airway Epithelial Cells"],"prefix":"10.31083","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0600-6836","authenticated-orcid":true,"given":"Mingzhi","family":"Luo","sequence":"first","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Zhang","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-1261-8534","authenticated-orcid":true,"given":"Kai","family":"Ni","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuanyu","family":"Shi","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-7516-266X","authenticated-orcid":true,"given":"Hang","family":"Li","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-2657-8483","authenticated-orcid":true,"given":"Lei","family":"Liu","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-2061-9534","authenticated-orcid":true,"given":"Yan","family":"Pan","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-1568-2499","authenticated-orcid":true,"given":"Jingjing","family":"Li","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1970-8239","authenticated-orcid":true,"given":"Linhong","family":"Deng","sequence":"additional","affiliation":[{"name":"Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, School of Medical and Health Engineering, School of Pharmacy, Changzhou University, 213164 Changzhou, Jiangsu, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,9,14]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","unstructured":"Madahar P, Beitler JR. 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