Soy Isoflavones Inhibit Both GPIb-IX Signaling and αIIbβ3 Outside-In Signaling via 14-3-3ζ in Platelet.

Ming Liu,Ren Lai,Chuanbin Shen,Heyu Ni,Runjia Xu,Gan Wang

doi:10.3390/molecules26164911

Abstract

Soy diet is thought to help prevent cardiovascular diseases in humans. Isoflavone, which is abundant in soybean and other legumes, has been reported to possess antiplatelet activity and potential antithrombotic effect. Our study aims to elucidate the potential target of soy isoflavone in platelet. The anti-thrombosis formation effect of genistein and daidzein was evaluated in ex vivo perfusion chamber model under low (300 s−1) and high (1800 s−1) shear forces. The effect of genistein and daidzein on platelet aggregation and spreading was evaluated with platelets from both wildtype and GPIbα deficient mice. The interaction of these soy isoflavone with 14-3-3ζ was detected by surface plasmon resonance (SPR) and co-immunoprecipitation, and the effect of αIIbβ3-mediated outside-in signaling transduction was evaluated by western blot. We found both genistein and daidzein showed inhibitory effect on thrombosis formation in perfusion chamber, especially under high shear force (1800 s−1). These soy isoflavone interact with 14-3-3ζ and inhibited both GPIb-IX and αIIbβ3-mediated platelet aggregation, integrin-mediated platelet spreading and outside-in signaling transduction. Our findings indicate that 14-3-3ζ is a novel target of genistein and daidzein. 14-3-3ζ, an adaptor protein that regulates both GPIb-IX and αIIbβ3-mediated platelet activation is involved in soy isoflavone mediated platelet inhibition.

Highlights

A systematic analysis for the Global Burden of Disease Study revealed that cardiovascular disease (CVD) has the highest mortality among all the non-communicable diseases [1].Thrombus is responsible for high-mortality CVD such as myocardial infarction and stroke, and platelets play important roles in the pathogenesis of these diseases [2,3,4]
(1800 s−1 ) in collagen-coated perfusion chamber. Both genistein (Figure 1A, CAS registry numbers are 446-72-0) and daidzein (Figure 1B, CAS registry numbers are 486-66-8) inhibited thrombi formation under the shear rate of 300 s−1 (Figure 1C,D) and 1800 s−1 (Figure 1E,F), while genistein showed higher potency. These isoflavones were preferred to inhibit the platelet aggregation and thrombosis at higher shear rate (1800 s−1 ), suggesting these compounds may have an inhibitory effect on platelet mechanosensor
To exclude integrin-mediated platelet aggregation, we examined the effect of the soy isoflavones on ristocetin-induced platelet agglutination in the presence of eptifibatide (Figure 2C) or EDTA

Summary

Introduction

A systematic analysis for the Global Burden of Disease Study revealed that cardiovascular disease (CVD) has the highest mortality among all the non-communicable diseases [1].Thrombus is responsible for high-mortality CVD such as myocardial infarction and stroke, and platelets play important roles in the pathogenesis of these diseases [2,3,4]. The inhibition of platelet function has been used for a long time to prevent and treat CVD [5]. The well-known correlation between diet and health provides strong evidence that functional foods may maintain or improve health and prevent CVD [6,7,8]. Epidemiological studies suggested that soy consumption is associated with a lower incidence of CVD [10,11,12]. Based on clinical trials and epidemiological data, the US Food and Drugs Administration approved a health claim for soy [13]. Dietary soy isoflavones rather than soy proteins exert antiplatelet functions in agonist-induced platelet activation [14,15]

Methods

Results

Conclusion