ABSTRACT:Bronchial wall remodeling is a major morbidity component in oxidant injury in bronchopulmonary dysplasia (BPD) and asthma. Hypothesis: IGF-1 enhances alpha smooth muscle expression and collagen synthesis in developing lung fibroblasts leading to fibrosis through nuclear NF-k B -dependent transcription. We studied NF-k B dependent transcription by transfecting HFLF with a NF-k B responsive promoter driving the luciferase gene and treating with IGF-1 (100 ng/mL) and measuring luciferase activity. We exposed cells to the PI-3 kinase inhibitor or the Erk1/2 inhibitor one hr before stimulating with IGF-1. We also used IGF-1 receptor antibody to inhibit the action of IGF-1 and studied its effect on alpha-sma and type I collagen. IGF-1 treatment significantly increased luciferase activity. This was attenuated by PI-3 kinase and MAP-Kinase inhibitors. Western blot analysis showed PI-3 kinase mediates IGF-1 activation of NF-k B independent of I K B phosphorylation. We found an up-regulation of phospho NF-kB in the nuclear extract compared with total NFKB showing that IGF-1 regulates NF-k B transcriptional activity downstream of NF-k B nuclear translocation. IGF-1-induced increase in alpha-sma expression and type-I collagen was significantly inhibited by pretreatment with LY294002 and IGF-1 receptor antibody. IGF-1 cell signaling leading to collagen synthesis in fetal lung fibroblasts is mediated by PI3 Kinase acting through NF-k B in HFLF. (Pediatr Res 60: 389-394, 2006) I n asthma, chronic inflammation leads to pathologic changes in the airways with marked remodeling (1). An increased number of myofibroblasts beneath the bronchial epithelial basement membrane has been described in asthma. Bronchial biopsies of asthmatic patients show thickening of the subepithelial layer due to the deposition of fibrillar collagen (2). The production of mediators by epithelial cells in close proximity to myofibroblasts during epithelial repair after repeated damage is one possible mechanism for airway remodeling (3). IGF-1 and IGF binding proteins (IGFBPs) are active in the pulmonary cellular environment to modulate cell proliferation and function (4). IGF-1 and its receptor, IGF-1R, are also known to modulate repair processes after lung injury (5). Others and we have observed that IGF-1R expression is more intense in areas of increased cell proliferation in developing lung (6,7). These studies prompted us to investigate the hypothesis that IGF-1 mediated signaling in developing lung cells may induce some of the phenotypic changes that are observed in asthma. We examined the signaling pathway activated by IGF-1 in fetal lung fibroblasts leading to an increase in myofibroblast proliferation and collagen synthesis, and whether this involves nuclear NF-kappaB (NF-k B)-dependent transcription. MATERIALS AND METHODSReagents were obtained as follows: Recombinant IGF-1 and IGF-1 receptor antibody were purchased from R&D Systems (Minneapolis, MN). Goat antirabbit IgG was purchased from Upstate Biotechnology (Charlottesville, VA). Penic...
We examined the cytoprotective effect of interleukin-6 (IL-6) and interleukin-11 (IL-11) during oxidant injury in neonatal lung and the regulators of cell death in vitro and in vivo after oxidant exposure. Type II cells from day 21 fetal neonatal rat lungs were treated with varying concentrations of either IL-6 or IL-11 for 24 hr prior to exposure to H(2)O(2). Three-day-old transgenic lung-specific IL-11 and IL-6 overexpressing and wild type (WT) mouse pups were exposed to hyperoxia or room air for 3 days. Type II cells exposed to either IL-6 or IL-11 prior to oxidant injury exhibited improved survival compared to controls, 67% +/- 2.6 survivals in IL-6 pretreated cells compared to 48% +/- 1.6 in control; 63% +/- 3 survivals in IL-11 pretreated cells compared to 49% +/- 2.6 in control. The number of TUNEL positive cells in hyperoxia-exposed lungs was increased compared to room air animals (27 +/- 0.9 vs. 4 +/- 0.4; mean +/- SEM; P < 0.05). In contrast, the number of TUNEL positive cells was reduced in hyperoxia-exposed lungs from IL-11 (+) mice (15.2 +/- 2.2; mean +/- SEM; P < 0.05). There was an enhanced accumulation of Bcl-2 and reduction of Bax protein in hyperoxia-exposed IL-11 (+) compared to room air-exposed mice. This was not seen in hyperoxia-exposed IL-6 (+) pups. An increase in caspase-3 was seen in hyperoxia-exposed lungs of WT pups compared to IL-11 (+) pups. IL-11 and IL-6 provide protective effects against oxidant-mediated injury in fetal type II cells and IL-11 provides protection in vivo by down-regulation of caspase-mediated cell death.
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