Nitric oxide-dependent activation of p53 suppresses bleomycin-induced apoptosis in the lung

Chronic inflammation leading to pulmonary fibrosis develops in response to environmental pollutants, radiotherapy, or certain cancer chemotherapeutic agents. We speculated that lung injury might be mediated by p53, a proapopt otic transcription factor widely implicated in the response of cells to DNA damage. Intratracheal administration of bleomycin led to caspase-mediated DNA fragmentation characteristic of apoptosis. The effects of bleomycin wer e associated with translocation of p53 from the cytosol to the nucleus only in alveolar macrophages that had been exposed to the drug in vivo, suggest ing that the lung microenvironment regulated p53 activation. Experiments wi th a thiol antioxidant (N-acetylcysteine) in vivo and nitric oxide (NO) don ors in vitro confirmed that reactive oxygen species were required for p53 a ctivation. A specific role for NO was demonstrated in experiments with indu cible nitric oxide synthase (iNOS)(-/-) macrophages, which failed to demons trate nuclear p53 localization after in vivo bleomycin exposure. Strikingly , rates of bleomycin-induced apoptosis were at least twofold higher in p53( -/-) C57BL/6 mice compared with heterozygous or wild-type littermates. Simi larly, levels of apoptosis were also twofold higher in the lungs of iNOS(-/ -) mice than were observed in wild-type controls. Consistent with a role fo r apoptosis in chronic lung injury, levels of bleomycin-induced inflammatio n were substantially higher in iNOS(-/-) and p53(-/-) mice compared with wi ld-type controls. Together, our results demonstrate that iNOS and p53 media te a novel apoptosis-suppressing pathway in the lung.