title: Up-regulation of inducible nitric oxide synthase in fibroblasts parallels the onset and progression of fibrosis in an experimental model of post-transplant obliterative airway disease.
abstract: The main cause of mortality following lung transplantation is chronic rejection, manifesting morphologically as obliterative bronchiolitis (OB). It has been suggested that damage to the respiratory epithelium initiates proliferation of mesenchymal cells, leading to dense collagenous scarring in small airways. Inducible nitric oxide synthase (iNOS) is strongly expressed in the damaged epithelium in human OB, along with high levels of peroxynitrite, suggesting that endogenous NO mediates the epithelial destruction. To examine further the role of iNOS in this process, heterotopic airway implants were studied in rats, an acknowledged disease model. Specimens of iso- or allografted trachea, collected 3-60 days after implantation, were processed for histology and immunocytochemistry for iNOS and, as a marker of peroxynitrite formation, nitrotyrosine. In both iso- and allografts at the earliest stage (day 3), ischaemia was associated with severe epithelial damage or loss. These changes progressed until day 7 and were accompanied by strong expression of iNOS and nitrotyrosine in epithelial cells. In isografts, epithelial recovery was seen, with abundant iNOS immunoreactivity but little nitrotyrosine. In contrast, the epithelium in allografts did not regenerate and progressive inflammation and fibroproliferation occurred until complete obliteration of the tracheal lumen at day 60. The fibroproliferation was associated with changes in morphology of fibroblasts that were accompanied by alterations in their iNOS expression. iNOS immunoreactivity was dense in the plump fibroblasts of early lesions, in some cases as early as post-operative day 5, but very weak in elongated fibroblasts in totally occluded grafts. The intensity of immunoreactivity for nitrotyrosine corresponded to that of iNOS. These results indicate a dual role for NO in the airway obliteration that follows transplantation, through destruction of epithelium and stimulation of fibroblast activity.
authors.affiliation: Department of Histochemistry, ICSM, Hammersmith Hospital, London, UK.
authors.forename: H M
authors.initials: HM
authors.lastname: Romanska
authors.forename: T S
authors.initials: TS
authors.lastname: Ikonen
authors.forename: A E
authors.initials: AE
authors.lastname: Bishop
authors.forename: R E
authors.initials: RE
authors.lastname: Morris
authors.forename: J M
authors.initials: JM
authors.lastname: Polak
mesh_terms: D000818:Animals; D001989:Bronchiolitis Obliterans / Q000201:enzymology* / Q000473:pathology; D018450:Disease Progression; D005347:Fibroblasts / Q000201:enzymology; D006084:Graft Rejection / Q000201:enzymology* / Q000473:pathology; D016040:Lung Transplantation / Q000473:pathology*; D008297:Male; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D011658:Pulmonary Fibrosis / Q000201:enzymology / Q000473:pathology; D051381:Rats; D011914:Rats, Inbred BN; D011917:Rats, Inbred Lew; D014132:Trachea / Q000201:enzymology; D015854:Up-Regulation
chemical_list: D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat