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. 2013 Sep;6(3):136-40.
doi: 10.2478/intox-2013-0021.

Hemolytic activity and platelet aggregation inhibitory effect of vipoxin's basic sPLA2 subunit

Affiliations

Hemolytic activity and platelet aggregation inhibitory effect of vipoxin's basic sPLA2 subunit

Silviya Stoykova et al. Interdiscip Toxicol. 2013 Sep.

Abstract

In the present study we evaluated the effect of secreted phospholipase A2 (sPLA2) (the toxic subunit of the heterodimeric neurotoxin vipoxin, isolated from the Bulgarian long-nosed viper Vipera ammodytes meridionalis) on hemolysis, erythrocyte morphology and platelet aggregation. Hemolytic activity of sPLA2 was examined in the presence of saturated (palmitic) and unsaturated (oleic) fatty acids and it was found that oleic acid increased the hemolytic activity of sPLA2 in a concentration-dependent manner, compared to the effect of palmitic acid and controls. The addition of heparin to red blood cells (RBC) suspension containing sPLA2 or mixture of sPLA2 and the corresponding fatty acid led to an inhibition of hemolytic activity. The effect of sPLA2 on RBC morphology resulted in formation of echinocytes (spherocyte subtype), suggesting that RBC could be the possible targets attacked by sPLA2. Vipoxin sPLA2 inhibited (in a dose-dependent manner) platelet aggregation when arachidonic acid and collagen were used as inducers, while in the case of ADP its inhibitory effect was inappreciable.

Keywords: hemolytic activity; platelet aggregation; secreted phospholipase A2; vipoxin.

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Figures

Figure 1
Figure 1
Effect of palmitic (a) and oleic (b) acids on RBC hemolysis: columns 1 – fatty acid only; columns 2 – fatty acid + heparin (17.5 IU/mL); columns 3 – fatty acid + sPLA2 (5 μg/mL); columns 4 – fatty acid + sPLA2 (5 μg/mL) + heparin (17.5 IU/mL). Results are reported as means ± SD (n = 5). The hydrolysis achieved by sPLA2 (5 μg/mL) is 1.12±0.34%; the hydrolysis in the presence of sPLA2 (5 μg/mL) and heparin (17.5 IU/mL) is 0.84 ± 0.37%.
Figure 2
Figure 2
Blood smears: normal erythrocytes (a); RBC treated with sPLA2 at concentrations of 0.2 μg/mL (b), 1 μg/mL (c), 5 μg/mL (d).
Figure 3
Figure 3
Effect of sPLA2 on inhibition of platelet aggregation induced by collagen (□しろいしかく, 87±9% aggregation), arachidonic acid (○しろまる, 80±8% aggregation) and ADP (Δ, 97±9% aggregation). Results are reported as means ± SD (n = 5).

References

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