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Protein, not adenosine or adenine nucleotides, mediates platelet decrease in endothelial permeability

S Patil¹, J E Kaplan, F L Minnear

¹Department of Physiology and Cell Biology, Albany Medical College, Union University, New York 12208, USA.

The American Journal of Physiology

|December 31, 1997

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Platelets reduce blood vessel leakiness via a protein, not adenosine. This study investigated platelet-conditioned medium (PCM) and found a protein, not adenosine, is responsible for decreasing endothelial permeability.

Area of Science:

Biochemistry
Cell Biology
Physiology

Background:

Platelets and platelet-conditioned medium (PCM) are known to decrease endothelial protein permeability in vitro.
Adenosine and a large protein (>100 kDa) were previously suggested as soluble factors responsible for this effect.

Purpose of the Study:

To investigate the specific role of adenosine in the platelet-mediated decrease of endothelial permeability.
To identify the soluble factors within PCM responsible for modulating endothelial barrier function.

Main Methods:

High-performance liquid chromatography (HPLC) was used to measure adenosine and adenine nucleotides (AMP, ADP, ATP) in PCM.
Endothelial permeability was assessed using 125I-labeled albumin clearance and electrical resistance across bovine pulmonary artery endothelial cell monolayers.

Related Experiment Videos

Experiments involved using adenosine deaminase and an adenosine-receptor antagonist to block adenosine's effects.

Fractionation of PCM based on molecular weight (<3 kDa and >3 kDa) and treatment with trypsin were employed.

Main Results:

PCM contained significant levels of AMP, ADP, and ATP, but adenosine was undetectable (<0.1 microM).
Neither adenosine deaminase nor an adenosine-receptor antagonist affected the PCM-induced decrease in endothelial permeability.
A protein fraction (>3 kDa) of PCM significantly decreased endothelial permeability, while a smaller fraction (<3 kDa) did not.
The permeability-reducing activity of PCM was sensitive to trypsin, indicating a proteinaceous nature.

Conclusions:

Adenosine and adenine nucleotides are not the primary factors mediating the platelet-induced decrease in endothelial permeability.
The results strongly suggest that a protein component within platelet-conditioned medium is responsible for reducing endothelial protein permeability.