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Am J Physiol Lung Cell Mol Physiol 260: L83-L89, 1991;
1040-0605/91 $5.00
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AJP - Lung Cellular and Molecular Physiology, Vol 260, Issue 2 83-L89, Copyright © 1991 by American Physiological Society

ARTICLES

Glucocorticoids induce neutral endopeptidase in transformed human tracheal epithelial cells

D. B. Borson and D. C. Gruenert
Department of Physiology, University of California, San Francisco 94143.

Neutral endopeptidase (NEP, also known as enkephalinase, CALLA, or EC 3.4.24.11) is a membrane-bound peptidase present in many different cell types. Previous studies have shown that it modulates the actions of a variety of biologically active peptides on several airway responses. More recent studies have demonstrated that reductions in neutral endopeptidase activity in animal airways is associated with increased responses to exogenously applied and endogenously released peptides. To study the regulation of NEP expression, we used human airway epithelial cells transformed in vitro with an origin-defective SV40 plasmid. Enzymatic activity, measured using [3H-Tyr,D-Ala2]leucine enkephalin, increased with cell density (1.4 ng/10(6) cells at 530 cells/cm2 and 21 ng/10(6) cells at confluence, 400 X 10(3) cells/cm2). In both confluent and nonconfluent cultures, the glucocorticoid budesonide increased neutral endopeptidase activity in time- and concentration-dependent fashions. Maximal increases of 10 ng/10(6) cells greater than control were observed after 6 days of incubation at 10(-7) M budesonide. Dexamethasone also increased NEP, suggesting that the effect is due to glucocorticoid receptor effects. Transcription, as assessed by Northern blot analysis of total cellular RNA, showed that NEP-specific RNAs also increased with increasing concentration of glucocorticoid. We conclude that neutral endopeptidase can be increased by cell growth or density and by glucocorticoids and that the effects of glucocorticoids are mediated by increased NEP gene expression.


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