Calcium mobilization by the plant estrogen ferutinin does not induce blood platelet aggregation

Abstract

Platelet activation is closely associated with an increase in intracellular Ca(2+) concentration. Various compounds including Ca(2+) ionophores are able to trigger platelet aggregation by increasing intracellular Ca(2+) concentration in platelets. In the present study, we monitored the effect of the phytoestrogen ferutinin, which acts as a Ca(2+) ionophore in human blood platelets; its ionophore-like properties include upregulation of [Ca(2+)](in), activation of fibrinogen receptors and increased fibrinogen binding. Using spectrofluorometry and triple-color flow cytometry, we demonstrate that ferutinin increases [Ca(2+)](in) in both isolated platelets and platelets in whole blood from humans. This effect was almost completely blocked by the Ca(2+) chelator EGTA and was not sensitive to either Gd(3+) or econazole, which inhibit VOC and SOC channels, respectively. Nor was the effect sensitive to thapsigargin, an inhibitor of endoplasmic reticulum Ca(2+) ATPases. Ferutinin stimulated the expression of the active form of the GPIIb-IIIa complex and whole blood platelet aggregation only weakly and had no statistically significant effect on the binding of fibrinogen. These results demonstrate apparently inconsistent effects of ferutinin, which raises intraplatelet Ca(2+) concentration but fails to have an effect on spontaneous blood platelet aggregation. This pattern of responses may be caused by the combination of ferutinin's Ca(2+) ionophoric and estrogenic properties.

Anti-thrombotic effects of selective estrogen receptor modulator tamoxifen

“…In agreement with this, earlier studies have demonstrated enhanced [Ca 2+ i ] mobilisation by tamoxifen in platelets stimulated with thrombin (30), Chinese hamster ovary cells (60) and cancer cell lines (61). Although inhibitory action of tamoxifen on platelet aggregation contradicts rise in [Ca 2+ i ], similar observation has also been documented with plant estrogen feruti-nin, which mobilised intracellular calcium in platelets without inducing platelet aggregation (62). Contrarily, a recent report by Chang et al (18) observed suppression of platelet cytosolic Ca 2+ by tamoxifen, which could be attributed to collagen, a different agonist, used by these authors.…”

Ferutinin Induces Membrane Depolarization, Permeability Transition Pore Formation, and Respiration Uncoupling in Isolated Rat Liver Mitochondria by Stimulation of Ca2+-Permeability

“…On the other hand, ferutinin acts as an electrogenic Ca 2+ -ionophore that has been shown to use planar lipid bilayers membranes and liposomes (Zamaraeva et al 1997;Abramov et al 2001;Dubis et al 2015). Similarly, earlier works showed that ferutinin at concentrations of 1-50 µM considerably increased Ca 2+ -permeability of thymocytes, platelets, Jurkat T-cell, erythrocytes, mitochondria, and sarcoplasmic reticulum (Zamaraeva et al 1997(Zamaraeva et al , 1999(Zamaraeva et al , 2010Macho et al 2004;Gao et al 2013). Ferutinin interacts strongly with Ca 2+ -ions via the hydroxyl group of ferutinol and the carboxyl oxygen of the complex ether bond (Dubis et al 2015).…”

“…Ferutinin was isolated from the roots of Ferula tenuisecta by the method of Saidkhodjaev and Nikonov (Saidkhodjaev and Nikonov 1973). According to the GC-MS analysis, the purity of ferutinin was 96 ± 2% (Zamaraeva et al 2010). We used ferutinin as 1 mM stock solution in DMSO.…”

“…In contrast, the stoichiometry of ferutinin-Ca 2+ complexes in the membrane was 1:1. It was demonstrated using Jurkat T-cells and erythrocytes, but not platelets, that an increase in cytosolic Ca 2+ at lower doses of ferutinin was connected with activation of P-and L-types of membranous Ca 2+ channels (Macho et al 2004;Zamaraeva et al 2010;Gao et al 2013). The study of Ca 2+ -ionophoric effects of ferutinin on neurons and hepatocytes demonstrated that increase in cytosolic Ca 2+ was accompanied by accumulation of Ca 2+ in mitochondria and, as a result, this was followed by depolarization of mitochondrial membranes, which could be prevented by cyclosporine A (CsA), an inhibitor of mitochondrial permeability transition (MPT) pore formation (Abramov and Duchen 2003).…”

Exploration of the Protective Effects of Some Natural Compounds against Neurodegeneration Exploiting Glycine Receptors in vivo Model

“…Several species of Ferula genus has been used in folk medicine in digestive disorders, rheumatism, headache, arthritis and as, antispasmodic and aphrodisiac [8]. Ferutinin (Jaeschkeanadiol p-hydroxy benzoate) ( Figure 1A) is mainly found in Ferula hermonis L. "Zallouh", and other Ferula species (like, Ferula tenuisecta) [9]. It belongs to a broad group of sesquiterpene ducane esters [10] and has a wide range of biological activities.…”