Kidney Res Clin Pract.
2017 Sep;36(3):224-231. 10.23876/j.krcp.2017.36.3.224.
A calpain inhibitor protects against fractalkine production in lipopolysaccharide-treated endothelial cells
- Affiliations
-
- 1Department of Microbiology, Chung-Ang University College of Medicine, Seoul, Korea.
- 2Department of Internal Medicine, Myongji Hospital, Seonam University College of Medicine, Goyang, Korea. intmdoh@hanmail.net
- KMID: 2416091
- DOI: http://doi.org/10.23876/j.krcp.2017.36.3.224
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)-α, interleukin (IL)-1, and interferon-γ. 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 μg/mL of LPS to determine cell viability. The changes of CX3CL1 expression were compared in control, LPS (1 μg/mL)-, IL-1α (1 μg/mL)-, and IL-1β (1 μg/mL)-treated HUVECs. Cell viability and CX3CL1 production were compared with 50 μM 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 μg/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-1β-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.
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