Selective Inhibition of P2Y1 and P2Y12 Receptor Signal Pathways in Platelet Aggregation in Transgenic Cell Lines and Rats by Potassium 2-(1-Hydroxypentyl)-Benzoate, Puerarin and Salvianolic Acid B

Abstract

Aim

Potassium 2-(1-hydroxypentyl)-benzoate (dl-PHPB), puerarin and salvianolic acid B are three natural products or derivatives that can inhibit platelet aggregation. However, the mechanisms of dl-PHPB, puerarin and salvianolic acid B to inhibit platelet aggregation are still not clear.

Method

Here, 2-methylthioadenosine diphosphate (2-MeSADP) was used as an inducer to confirm the effects of three drugs on platelet aggregation and illustrate the corresponding mechanisms.

Result

The results indicated that dl-PHPB, puerarin and salvianolic acid B significantly inhibited platelet aggregation both in vivo and in vitro. In addition, the content of IP₃, cAMP and intracellular [Ca²⁺]_i were measured in HEK293 cell lines overexpressing P2Y₁ and P2Y₁₂. Dl-PHPB and puerarin could obviously reduce 2-MeSADP-induced IP₃ increase, but salvianolic acid B showed no effects. Unlike dl-PHPB and puerarin, which had no effects on 2-MeSADP-induced cAMP decrease, salvianolic acid B significantly reversed the reduction of cAMP. Both dl-PHPB and puerarin could decrease the enhanced intracellular [Ca²⁺]_i induced by 2-MeSADP; however, salvianolic acid B showed no effect on intracellular [Ca²⁺]_i elevation.

Conclusion

These results suggested that dl-PHPB and puerarin inhibited platelet aggregation via targeting at P2Y₁ receptor and P2Y₁-Gq-IP₃-Ca²⁺ signal pathway. Differently, salvianolic acid B inhibited platelet aggregation via targeting at P2Y₁₂ receptor and via Gi-AC-cAMP signal pathway.