Sodium azide, a bacteriostatic preservative contained in commercially available laboratory reagents, influences the responses of human platelets via the cGMP/PKG/VASP pathway.
OBJECTIVE
The bacteriostatic preservative sodium azide (NaN(3)) activates soluble guanylate cyclase (sGC) in vascular tissues, thus elevating cellular 3',5'-cyclic guanosine monophosphate (cGMP). Because the sGC/cGMP pathway is involved in the control of platelet aggregation, we investigated whether in human platelets NaN(3) influences the responses to agonists, cGMP levels and cGMP-regulated pathways.


METHODS
Concentration- and time-dependent effects of NaN(3) (1-100 micromol/L; 5-60 min incubation) on ADP- and collagen-induced aggregation, NO synthase (NOS) activity, cGMP synthesis and vasodilator-stimulated phosphoprotein (VASP) phosphorylation at Ser239 were investigated in platelets from 21 healthy individuals.


RESULTS
NaN(3) exerted concentration- and time-dependent antiaggregatory effects starting from 1 micromol/L (IC(50) with 5-min incubation: 2.77+/-0.35 micromol/L with ADP and 4.64+/-0.48 micromol/L with collagen) and significantly increased intraplatelet cGMP levels and phosphorylation of VASP at Ser239 at 1-100 micromol/L; these effects were prevented by sGC inhibition, but not by NOS inhibition.


CONCLUSIONS
NaN(3) exerts antiaggregatory effects in human platelets via activation of the sGC/cGMP/VASP pathway. This biological effect must be considered when azide-containing reagents are used for in vitro studies on platelet function.
Russo
Isabella
I
Diabetes Unit, Department of Clinical and Biological Sciences of The University of Turin, San Luigi Gonzaga Hospital, Orbassano (Turin), Italy.
Del Mese
Paola
P
Viretto
Michela
M
Doronzo
Gabriella
G
Mattiello
Luigi
L
Trovati
Mariella
M
Anfossi
Giovanni
G
D000328:Adult; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D006152:Cyclic GMP / Q000378:metabolism*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism*; D005260:Female; D006801:Humans; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D019810:Sodium Azide / Q000494:pharmacology*; D000098662:Vasodilator-Stimulated Phosphoprotein
D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D010750:Phosphoproteins; D000098662:Vasodilator-Stimulated Phosphoprotein; D009569:Nitric Oxide; D019810:Sodium Azide; D017869:Cyclic GMP-Dependent Protein Kinases; D006152:Cyclic GMP