Previous studies suggested an association between changes in extracellular matrix (ECM) and pulmonary epithelial cell growth and differentiation in lungs of adrenalectomized rats. The role of adrenal hormones to modulate synthesis of cell and ECM proteins by type II epithelial cells was thus investigated. In cells isolated from lungs of adrenalectomized animals, synthesis of ECM proteins was stimulated to a greater extent than was synthesis of cell proteins. This resulted in increased relative synthesis of matrix components. Conversely, dexamethasone (200 nM) treatment in vitro reduced the effects of adrenalectomy and, in control cells, decreased relative ECM synthesis. The magnitude of these effects depended on time in culture and serum conditions. For example, culture of normal cells in serum-free medium mimicked the effects of adrenalectomy, whereas there were no effects of dexamethasone in the presence of serum. These results indicate a role for adrenal steroid hormones to modulate ECM metabolism by type II pulmonary epithelial cells and suggest that consequent effects on ECM biological activity may modify epithelial cell differentiation or growth.