Reactive oxygen and nitrogen intermediates increase transforming growth factor-beta1 release from human epithelial alveolar cells through two different mechanisms.
Transforming growth factor (TGF)-beta1 is a growth factor involved in the mechanisms of lung repair and fibrosis that follow inflammatory processes. We sought to examine the link between the generation of reactive oxygen intermediates (ROI) or reactive nitrogen intermediates (RNI) by inflammatory cells and the expression of TGF-beta1 by alveolar epithelial cells. Exposure of the A549 lung epithelial cell line to either an ROI generating system (xanthine and xanthine oxidase) or an RNI donor (S-nitroso-N-acetyl-penicillamine [SNAP]) promoted a time- and dose-dependent increase in TGF-beta1 release, as measured by a specific enzyme-linked immunosorbent assay. At the peak, the levels of TGF-beta1 were twice the control values. The induction of TGF-beta1 release by ROI was blunted by catalase and unaffected by superoxide dismutase, indicating the involvement of hydrogen peroxide. The response was also blunted by 5, 6-dichloro-1-beta-D-ribofuranosyl benzimidazole (DRB), a specific RNA polymerase II inhibitor, and accompanied by a corresponding increase in TGF-beta1 messenger RNA, as measured by quantitative/competitive reverse transcription polymerase chain reaction, suggesting the involvement of transcriptional mechanisms and possibly other downstream mechanisms. In contrast, RNI-induced TGF-beta1 release was unaffected by DRB and blunted by the protein synthesis inhibitor cycloheximide, suggesting the involvement of translational and post-translational mechanisms. This response required cyclic guanosine monophosphate (cGMP)- mediated processes because (1) immunoreactive cGMP accumulated in the culture medium of SNAP-treated cells; (2) SNAP-induced TGF-beta1 release was blunted by KT 5823, an inhibitor of cGMP-dependent protein kinase; and (3) similar increase in TGF-beta1 release was obtained by cell exposure to membrane-permeable dibutyryl-cGMP or to atrial natriuretic factor, a known agonist of particulate guanylate cyclase. These data suggest that in vitro exposure of human alveolar epithelial cells to ROI and RNI enhances TGF-beta1 release through different mechanisms. In vivo, this control may constitute a molecular link between inflammatory and fibrotic processes.
Bellocq
A
A
Service d'Explorations Fonctionnelles and Unité INSERM 489, Service de Biostatistique et Informatique, and Service de Pneumologie and UPRES-A 1531, Hôpital Tenon, UPRES-A 8068, Hôpital Cochin, Paris, France.
Azoulay
E
E
Marullo
S
S
Flahault
A
A
Fouqueray
B
B
Philippe
C
C
Cadranel
J
J
Baud
L
L
D001120:Arginine / Q000494:pharmacology; D009320:Atrial Natriuretic Factor / Q000494:pharmacology; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000378:metabolism; D016166:Free Radical Scavengers / Q000494:pharmacology; D006157:Guanosine Monophosphate / Q000502:physiology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009584:Nitrogen / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011650:Pulmonary Alveoli / Q000187:drug effects* / Q000502:physiology*; D012323:RNA Processing, Post-Transcriptional; D017382:Reactive Oxygen Species; D013482:Superoxide Dismutase / Q000494:pharmacology; D013997:Time Factors; D014158:Transcription, Genetic; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000494:pharmacology
D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D011500:Protein Synthesis Inhibitors; D017382:Reactive Oxygen Species; D016212:Transforming Growth Factor beta; D004004:Dichlororibofuranosylbenzimidazole; D006157:Guanosine Monophosphate; D009320:Atrial Natriuretic Factor; D001120:Arginine; D003513:Cycloheximide; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D009584:Nitrogen; D019331:NG-Nitroarginine Methyl Ester