The inhibitory effect of cinchonine on human platelet aggregation due to blockade of calcium influx

Abstract

The Cinchona bark contains alkaloids like quinine, quinidine, cinchonine and cinchonidine. These agents are effective antimalarial drugs and have been used clinically in malaria caused by Plasmodium falciparum. Previous studies show that quinine and quinidine exert effects on cardiovascular system. This study was conducted to examine the effect of cinchonine on human platelet aggregation. The results show that cinchonine inhibited platelet aggregation mediated by platelet agonists, epinephrine (200 μM), ADP (4.3 μ M), platelet activating factor (PAF; 800 nM) and collagen (638 nM) but had no effect on arachidonic acid (AA; 0.75 mM). Cinchonine was most effective in inhibiting aggregation induced by platelet activating factor and epinephrine with ic 50 values of 125 and 180 μM respectively, however, higher concentrations of cinchonine were required to inhibit aggregation mediated by ADP or collagen ( ic 50; 300 μM). Pretreatment of platelets with cinchonine inhibited aggregation caused by Ca 2+ ionophore, A-23187 (6 μM), in a dose-dependent manner ( ic 50; 300 μM) indicating an inhibitory effect on Ca 2+-signaling cascade. This was supported by measuring [Ca 2+]i in platelets loaded with Fura-2AM where cinchonine inhibited the rise in cytosolic Ca 2+ mediated by A-23187 (6 μM) or collagen (638 nM). Results show that cinchonine (20 μM) also inhibited aggregation when platelets were pretreated with protein kinase C (PKC) activator, phorbol myristate acetate (PMA; 0.1 μM) in combination with low doses of platelet activating factor (80 nM). Cinchonine, however, had no effect on AA-induced platelet aggregation and thromboxane A 2 (TXA 2) synthesis in platelets. These results suggest that antiplatelet effects of cinchonine are mediated mainly through inhibition of Ca 2+-influx and protein kinase C pathways in platelets.