Ethanol-exposed lung fibroblasts cause airway epithelial barrier dysfunction.
Alcohol Clin Exp Res (Hoboken)
; 47(10): 1839-1849, 2023 Oct.
Article
en En
| MEDLINE
| ID: mdl-37864530
ABSTRACT
BACKGROUND:
Chronic alcohol ingestion predisposes to lung injury and disrepair during sepsis. Our previous studies outlined roles for transforming growth factor-beta 1 (TGFß1) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in epithelial barrier homeostasis and how alcohol perturbs their expression and signaling. Here we hypothesize that ethanol-exposed lung fibroblasts (LF) are a source of dysregulated TGFß1 and GM-CSF and thereby alter airway epithelial barrier function.METHODS:
Human or rat LF were cultured ± ethanol for 2 weeks and then co-cultured with human or rat airway epithelial cells (AEC) seeded on Transwell permeable supports. In selected groups, a TGFß1 receptor type 1 (TGFßR1) inhibitor (SB431542) or a TGFß1 neutralizing antibody was applied. Transepithelial electrical resistance (TER) was measured prior to co-culture and on day 5 of co-culture. AEC were then analyzed for the expression of selected tight junction and mesenchymal proteins, and transwell membranes were analyzed by immunofluorescence microscopy for ZO-1 expression and localization. TGFß1 and GM-CSF levels in conditioned media from the co-cultures were quantified by ELISA.RESULTS:
AEC co-cultured with ethanol-exposed LF (ELF) showed a significant reduction in TER and corresponding decreases in ZO-1 expression, whereas collagen type 1A1 and α-smooth muscle actin protein expression were increased. In parallel, in conditioned media from the ELF + AEC co-cultures, activated TGFß1 levels increased and GM-CSF levels decreased. Notably, all the effects of ELF on the AEC were prevented by blocking TGFß1 activity.CONCLUSIONS:
Prior ethanol exposure to LF induces barrier dysfunction in naive AEC in a paracrine fashion through activation of TGFß1 signaling and suppression of GM-CSF. These experimental findings provide a potential mechanism by which chronic alcohol ingestion impairs airway epithelial integrity and renders individuals susceptible to lung injury.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Idioma:
En
Revista:
Alcohol Clin Exp Res (Hoboken)
Año:
2023
Tipo del documento:
Article
País de afiliación:
Estados Unidos