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The peptidase inhibitor CGS‐26303 increases endothelin converting enzyme‐1 expression in endothelial cells through accumulation of big endothelin‐1
Author(s) -
Raoch V,
MartinezMiguel P,
ArribasGomez I,
RodriguezPuyol M,
RodriguezPuyol D,
LopezOngil S
Publication year - 2007
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1038/sj.bjp.0707398
Subject(s) - phosphoramidon , thiorphan , bosentan , endothelin 1 , neprilysin , endothelin receptor , cycloheximide , endothelins , medicine , endothelin 3 , endocrinology , chemistry , biology , microbiology and biotechnology , receptor , enzyme , biochemistry , protein biosynthesis
Background and purpose: CGS‐26303 inhibits endothelin converting enzyme (ECE)‐1 more specifically than phosphoramidon. We have studied the effect of CGS‐26303 on ECE‐1 expression in bovine aortic endothelial cells. Methods: ECE‐1 activity and big endothelin (ET)‐1 levels were measured by ELISA, ECE‐1 expression using western and northern blot and promoter activity using transfection assays. Key results: ECE‐1 activity was completely inhibited by CGS‐26303 25 μ M and phosphoramidon 100 μ M . CGS‐26303 and phosphoramidon, though not thiorphan, a neutral endopeptidase (NEP) inhibitor, stimulated ECE‐1 expression in cells (maximal effect at 16 h, 25 μ M ). Cycloheximide abolished that effect. CGS‐26303 induced ECE‐1 mRNA expression and ECE‐1 promoter activity. CGS‐35066, a selective ECE‐1 inhibitor, mimicked the effects of CGS‐26303, suggesting that the effect was specific to ECE‐1 inhibition. Big ET‐1 accumulated in the cells and in the supernatants after CGS‐26303 treatment. Neither exogenously added ET‐1 nor the blockade of their receptors with bosentan modified ECE‐1 protein. When big ET‐1 was added to cells, significant increases in ECE‐1 protein content and ECE‐1 promoter activity were found. Bosentan did not block those effects. CGS‐26303 did not modify prepro‐ET‐1 expression. CGS‐26303 and big ET‐1 induced the same effects in human endothelial cells, at lower doses. Conclusions: These results suggest that the accumulation of big ET‐1 is responsible for the effects of CGS‐26303 on ECE‐1 and they did not depend on NEP blockade. Changes in ECE‐1 protein after the administration of CGS‐26303 could lead to a decreased response in long‐term treatments. British Journal of Pharmacology (2007) 152 , 313–322; doi: 10.1038/sj.bjp.0707398 ; published online 23 July 2007