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Updated: Jul 22, 2026
Analysis of Physiologic E-Selectin-Mediated Leukocyte Rolling on Microvascular Endothelium
Published on: February 11, 2009
Delayed treatment with diethyl maleate prevents E-selectin expression in human endothelial cells
Background:
Polymorphonuclear leukocyte (PMN) infiltration is a significant contributor to tissue damage in many disease states and is known to occur through an orderly set of events. The endothelial cell adhesion molecule E-selectin is involved in the initial rolling of PMNs on the endothelium at sites of inflammation. We have previously shown that the glutathione depleting agent diethyl maleate (DEM) attenuates lung injury in a rodent model of intratracheal LPS stimulation. We hypothesized that DEM might attenuate E-selectin in LPS-treated human umbilical vein endothelial cells as a mechanism underlying this effect. Further, we investigated the role of delayed treatment with DEM on E-selectin expression.
Methods:
Human umbilical vein endothelial cells were treated with DEM (100 to 400 mumol/L) before or after LPS stimulation (1 microgram/mL). Surface expression of E-selectin was examined using a cellular enzyme-linked immunosorbent assay. E-selectin mRNA transcripts were detected by Northern blot analysis. Nuclear factor-kappa B (NF-kappa B) activity was detected with gel shift assays.
Results:
DEM significantly inhibited LPS-induced E-selectin surface expression and mRNA levels in a dose-dependent fashion, with complete inhibition at 250 mumol/L, without affecting cell viability. This inhibitory effect was seen even if DEM was added up to 60 minutes after LPS. DEM inhibited NF-kappa B nuclear translocation in a manner that mirrored protein and mRNA levels.
Conclusions:
Delayed treatment with DEM attenuates NF-kappa B nuclear translocation and E-selectin expression in human umbilical vein endothelial cells up to 60 minutes after the onset of LPS stimulation. Thus, DEM may represent an effective intervention for PMN-mediated organ injury even when given after an inflammatory insult.
Insights
Diethyl maleate (DEM) reduces inflammation by inhibiting E-selectin and NF-kappa B activation in endothelial cells. This suggests DEM can treat polymorphonuclear leukocyte-mediated organ injury even after an inflammatory insult.
Area of Science:
- Endothelial cell biology
- Inflammation research
- Molecular medicine
Background:
- Polymorphonuclear leukocyte (PMN) infiltration contributes to tissue damage.
- E-selectin mediates initial PMN rolling on endothelium during inflammation.
- Diethyl maleate (DEM) previously showed potential in attenuating lung injury.
Purpose of the Study:
- To investigate if DEM attenuates E-selectin in LPS-treated endothelial cells.
- To explore the mechanism of DEM's anti-inflammatory effects.
- To determine the efficacy of delayed DEM treatment on E-selectin expression.
Main Methods:
- Human umbilical vein endothelial cells treated with DEM before or after LPS stimulation.
- E-selectin surface expression measured by ELISA.
- E-selectin mRNA and NF-kappa B activity analyzed via Northern blot and gel shift assays.
Main Results:
- DEM dose-dependently inhibited LPS-induced E-selectin surface expression and mRNA.
- Complete inhibition of E-selectin achieved at 250 mumol/L DEM without affecting cell viability.
- DEM inhibited NF-kappa B nuclear translocation, mirroring effects on E-selectin levels.
Conclusions:
- Delayed DEM treatment effectively reduces NF-kappa B and E-selectin expression post-LPS.
- DEM shows promise as an intervention for PMN-mediated organ injury, even when administered after inflammation onset.
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