Triflavin inhibits platelet-induced vasoconstriction in de-endothelialized aorta

Triflavin, a 7.5-kD cysteine-rich polypeptide purified from Trimeresurus favoviridis snake venom, belongs to a family of Arg-Gly-Asp-(RGD)-containing peptides, termed disintegrins. In this study, aggregating human platelets dose-dependently induced vasoconstriction in de-endothelialized isolated rat thoracic aortas. At 5x10(7) cells per milliliter, platelets induced a peak tension averaging 65+/-7.2% of the tension induced by phenylephrine (10 mu mol/L). The relative effectiveness of RGD-containing peptides (including venom peptides triflavin and trigramin, small RGD synthetic peptides Gly-Arg-Gly-Asp-Ser [GRCDS], Gly-Arg-Gly-Asp-Phe [GRGDF], and Gly-Arg-Gly-Asp-Ser-Pro-Lys [GRGDSPK]) was examined by testing the inhibitory effect on aggregating platelet-induced vasoconstriction in deendothelialized aorta. Triflavin (1 mu mol/L) significantly inhibited the platelet-induced vasoconstriction, whereas neither trigramin (10 mu mol/L) nor small RGD peptides (2 mmol/L) (ie, GRGDS, GRGDF, and GRGDSPK) showed any significant effect, The release of serotonin and the formation of thromboxane A(2) from aggregating platelets were both significantly inhibited by triflavin (2 mu mol/L), whereas trigramin and small RGD-containing peptides showed no significant. effect. On scanning electron micrographs of de-endothelialized aorta, aggregating platelets adhered to the subendothelium, with loss of their discoid shape. to form irregular spheres with pseudopod extensions. Triflavin (2 mu mol/L) markedly reduced the adhesion of platelets to the subendothelium in the same aorta. Furthermore, RGD-containing peptides (including triflavin, trigramin, and small RGD-containing peptides) inhibited the adhesion of 10 mu g/mL collagen-activated platelets to extracellular matrices (iel fibronectin, vitronectin, and von Willebrand factor). It is concluded that the marked ability of triflavin to inhibit aggregating platelet-induced vasoconstriction in de-endothelialized aorta compared with other RGD-containing peptides (including trigramin), may be due at least, partly to triflavin's efficiently preventing the activation of platelets subsequent to inhibition of serotonin release and thromboxane A(2) formation. However. the different abilities of triflavin compared with other RGD-containing peptides was not related to the ability to inhibit adhesion of placelets to extracellular matrices. Therefore, from the results of this study, it appears that triflavin may be a useful therapeutic agent for the treatment of thromboembolism and its associated angiospasm.