Neutral/Alkaline and Acid Ceramidase Activities Are Actively Released by Murine Endothelial Cells

• DOI: 10.1006/bbrc.2000.3370
• Publication Date: 2002-09-16T14:49:19Z
• Authors: Elena Romiti; Elisabetta Meacci; Motohiro Tani; Francesca Nuti; Marta Farnararo; Makoto Ito; Paola Bruni
• Keywords: 0301 basic medicine; 0303 health sciences; Brefeldin A; Time Factors; Acid Ceramidase; Macrophages; Fibroblasts; Hydrogen-Ion Concentration; Bradykinin; Amidohydrolases; Cell Line; Mice; 03 medical and health sciences; Culture Media, Conditioned; Ceramidases; Dactinomycin; Animals; Humans; Tetradecanoylphorbol Acetate; Endothelium; Cycloheximide; Interleukin-1

Ceramidases (CDase(s)) play a key role in sphingolipid metabolism by hydrolyzing ceramide into sphingosine. Here we report that murine endothelial cells, macrophages, and human fibroblasts are all able to release acid as well as neutral/alkaline CDase activities in the culture medium. Endothelial cells were characterized by the highest specific activity of cellular as well as secreted CDases. The release of both enzymatic activities was reduced by protein synthesis inhibitor cycloheximide but was unaffected by the blocking of RNA transcription with actinomycin D. The discharge of acid and neutral/alkaline CDases was also diminished by brefeldin A, a fungal metabolite which disrupts Golgi apparatus. Remarkably, treatment of endothelial cells with bradykinin resulted in a significant increase of neutral/alkaline but not acid CDase release. This report represents the first evidence for the existence of constitutive and regulated release of CDase activities by endothelial cells. In view of the known ability of these cells to secrete sphingomyelinase, this finding suggests that CDase may participate in extracellular sphingomyelin metabolism which is presently known to have a role in atherogenesis and could be involved in other physiological or pathological events.