The Acute Respiratory Distress Syndrome (ARDS) is a common form of acute lung injury with no effective therapy and mortality of 50%. Investigators in this Center application demonstrated that chronic alcohol abuse was the first co-morbid variable to be identified that significantly increases the incidence and severity of ARDS. In this Center application, the mechanisms by which chronic ethanol ingestion predisposes to sepsis-induced acute lung injury will be addressed in alveolar endothelial cells, alveolar epithelial cells, pulmonary fibroblasts, and alveolar macrophages isolated from a rat model of chronic ethanol ingestion. When the role of glutathione availability in ethanol-induced toxicity is the study question, the rats will be fed the glutathione precursors N-acetylcysteine or Procysteine (L-2-oxo-4-thiazolidine-carboxylic acid) either during chronic ethanol ingestion. The purpose of the cell culture core will be to isolate and culture these cells after the appropriate dietary regimen and then distribute the cells to the different projects. For Projects 1 and 2 and Pilot Project 2, the cell culture core will provide primary alveolar type II cells. For Project 1, alveolar macrophages will also be provided. For Project 3 and Pilot Project 2, the core will provide pulmonary fibroblasts and alveolar type II cells from control or ethanol-exposed rats. For Projects 4 and 5 and Pilot Project 2, the core will be responsible for the isolation and culture of pulmonary microvascular endothelial cells. Peripheral neutrophils will also be provided for Project 4. This core will provide cells with maximum reproducibility, efficiency and cost effectiveness. These primary cells will be isolated and cultured in strict endotoxin-free conditions by experienced cell culture technicians. The purity of the cells obtained will routinely be verified using immunohistochemical techniques and functional assays. Cultured cells will be distributed to all investigators according to their requests for studies outlined in their proposals.
Showing the most recent 10 out of 104 publications
