Bleomycin-induced
lung injury is characterized in the neonatal rat by
inflammation, arrested lung growth, and
pulmonary hypertension (PHT), as observed in human infants with severe
bronchopulmonary dysplasia. Inhalation of CO(2) (therapeutic
hypercapnia) has been described to limit
cytokine production and to have anti-inflammatory effects on the injured lung; we therefore hypothesized that therapeutic
hypercapnia would prevent
bleomycin-induced
lung injury. Spontaneously breathing rat pups were treated with
bleomycin (1 mg/kg/d ip) or saline vehicle from postnatal days 1-14 while being continuously exposed to 5% CO(2) (Pa(CO(2)) elevated by 15-20 mmHg), 7% CO(2) (Pa(CO(2)) elevated by 35 mmHg), or normocapnia.
Bleomycin-treated animals exposed to 7%, but not 5%, CO(2), had significantly attenuated lung tissue macrophage influx and PHT, as evidenced by normalized pulmonary vascular resistance and right ventricular systolic function, decreased
right ventricular hypertrophy, and attenuated remodeling of pulmonary resistance arteries. The level of CO(2) neither prevented increased tissue neutrophil influx nor led to improvements in decreased lung weight, septal thinning, impaired alveolarization, or decreased numbers of peripheral arteries.
Bleomycin led to increased expression and content of lung
tumor necrosis factor (TNF)-α, which was found to colocalize with tissue macrophages and to be attenuated by exposure to 7% CO(2). Inhibition of TNF-α signaling with the soluble TNF-2 receptor
etanercept (0.4 mg/kg ip from days 1-14 on alternate days) prevented
bleomycin-induced PHT without decreasing tissue macrophages and, similar to CO(2), had no effect on arrested alveolar development. Our findings are consistent with a preventive effect of therapeutic
hypercapnia with 7% CO(2) on
bleomycin-induced PHT via attenuation of macrophage-derived TNF-α. Neither tissue macrophages nor TNF-α appeared to contribute to arrested lung development induced by
bleomycin. That 7% CO(2) normalized pulmonary vascular resistance and right ventricular function without improving inhibited airway and vascular development suggests that vascular hypoplasia does not contribute significantly to functional changes of PHT in this model.