A calpain inhibitor protects against fractalkine production in lipopolysaccharide-treated endothelial cells

Abstract

Background: Fractalkine (CX3CL1) is a chemokine with a unique CX3C motif and is produced by endothelial cells stimulated with lipopolysaccharide (LPS), tumor necrosis factor (TNF)-alpha, interleukin (IL)-1, and interferon-gamma. There have been several reports that the caspase/calpain system is activated in endotoxemia, which leads to cellular apoptosis and acute inflammatory processes. We aimed to determine the role of the caspase/calpain system in cell viability and regulation of fractalkine production in LPS-treated endothelial cells. Methods: Human umbilical vein endothelial cells (HUVECs) were stimulated with 0.01-100 mu g/mL of LPS to determine cell viability. The changes of CX3CL1 expression were compared in control, LPS (1 mu g/mL)-, IL-1 alpha (1 mu g/ mL)-, and IL-1 beta (1 mu g/mL)-treated HUVECs. Cell viability and CX3CL1 production were compared with 50 mu M of inhibitors of caspase-1, caspase-3, caspase-9, and calpain in LPS-treated HUVECs. Results: Cell viability was significantly decreased from 1 to 100 mu g/mL of LPS. Cell viability was significantly restored with inhibitors of caspase-1, caspase-3, caspase-9, and calpain in LPS-treated HUVECs. The expression of CX3CL1 was highest in IL-1 beta-treated HUVECs. CX3CL1 production was highly inhibited with a calpain inhibitor and significantly decreased with the individual inhibitors of caspase-1, caspase-3, and caspase-9. Conclusion: The caspase/calpain system is an important modulator of cell viability and CX3CL1 production in LPS-treated endothelial cells.