| 19115040 |
Albrecht EA, Sarma JV, Ward PA: Activation by C5a of endothelial cell caspase 8 and cFLIP. Inflamm Res. 2009 Jan;58(1):30-7.
OBJECTIVES AND DESIGN: In this study, we examine the relationship between C5a and activation of cysteine aspartic acid protease 8 (caspase 8) in human umbilical vein endothelial cells (HUVEC). MATERIALS OR SUBJECTS: Primary cultures of HUVEC were used. TREATMENTS: Recombinant human C5a (50 ng/ml) was used in the presence or absence of 10 microg/ml cycloheximide (CHX). METHODS: HUVEC were treated with C5a alone and in the presence of CHX, then monitored for cell viability, poly- ADP-ribose 1 (PARP-1) and caspase 8 activities. Gene and protein expressions were assessed for caspase 8 and the caspase 8 homologue, FLICE -inhibitory protein (cFLIP). RESULTS: We found a 43.1 +/- 6.9 percent reduction in viability of HUVEC stimulated for 18 h with 50 ng/ml C5a in the presence of 10 microg/ml CHX (p < 0.05). In contrast, the cell viability of cells stimulated for 18 h with 50 ng/ml C5a or 10 microg/ml CHX alone was not significantly different compared to the non-stimulated control. Treatment of HUVEC with C5a induced an increase in caspase 8 activity but did not significantly affect cFLIP levels. CONCLUSIONS: These data suggest caspase 8 activation induced by C5a leads to cell death if protein synthesis of antiapoptotic protein (s) is blocked. |
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