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Platelet-activating Factor (PAF) Increases NO Production in Human Endothelial Cells-Real-Time Monitoring by DAR-4M AM

Platelet-activating Factor (PAF) Increases NO Production in Human Endothelial Cells-Real-Time Monitoring by DAR-4M AM

M Kikuchi, H Shirasaki, T Himi

Prostaglandins Leukot Essent Fatty Acids. Apr-May 2008;78(4-5):305-9.

PMID: 18502110

Abstract:

Background:
Platelet-activating factor (PAF) is a potent inflammatory lipid mediator that increases vascular permeability and vasodilation. Several studies have addressed the effect of PAF on nitric oxide (NO) production from microvessels in vivo.
Objective:
The aim of present study was to evaluate the effect of PAF on NO production in primary cultured human vascular endothelial cells.
Methods:
Human umbilical vein endothelial cells (HUVECs) were loaded with diaminorhodamine-4M acetoxymethyl ester (DAR-4MAM), and the cells were stimulated with PAF. Intracellular NO production was monitored as increase in fluorescence intensity. Also, NO production was visualized at cellular levels using DAR-4M AM and fluorescence imaging.
Results:
Significant increases in NO production in HUVECs were soon after the PAF stimulation, reaching a plateau after 10 min of the stimulation. The increase of NO production at 10 min after the stimulation was statistically significant (p<0.05) for 0.01-10 microM PAF. PAF-induced NO production was abolished by pretreatment of HUVECs with a NOS inhibitor N(G)-monomethyl-L-arginine (L-NMMA) or PAF receptor antagonist BN 52021. LysoPAF, the inactive metabolite of PAF, did not exert a significant effect on intracellular NO levels.
Conclusions:
These results provide direct evidence that PAF cause intracellular NO production via activation of PAF receptors in human vascular endothelial cells.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42418052	DAR-4M AM			Price

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