Salidroside inhibits platelet function and thrombus formation through AKT/GSK3β signaling pathway

Abstract

Salidroside is the main bioactive component in Rhodiola rosea and possesses multiple biological and pharmacological properties. However, whether salidroside affects platelet function remains unclear. Our study aims to investigate salidroside's effect on platelet function. Human or mouse platelets were treated with salidroside (0-20 μM) for 1 hour at 37°C. Platelet aggregation, granule secretion, and receptors expression were measured together with detection of platelet spreading and clot retraction. In addition, salidroside (20 mg/kg) was intraperitoneally injected into mice followed by measuring tail bleeding time, arterial and venous thrombosis. Salidroside inhibited thrombin-or CRP-induced platelet aggregation and ATP release and did not affect the expression of P-selectin, glycoprotein (GP) Ibα, GPVI and αIIbβ3. Salidroside-treated platelets presented decreased spreading on fibrinogen or collagen and reduced clot retraction with decreased phosphorylation of c-Src, Syk and PLCγ2. Additionally, salidroside significantly impaired hemostasis, arterial and venous thrombus formation in mice. Moreover, in thrombin-stimulated platelets, salidroside inhibited phosphorylation of AKT (T308/S473) and GSK3β (Ser9). Further, addition of GSK3β inhibitor reversed the inhibitory effect of salidroside on platelet aggregation and clot retraction. In conclusion, salidroside inhibits platelet function and thrombosis via AKT/GSK3β signaling, suggesting that salidroside may be a novel therapeutic drug for treating thrombotic or cardiovascular diseases.

Matrine Impairs Platelet Function and Thrombosis and Inhibits ROS Production

“…Platelet aggregation was conducted in the presence of fibrinogen (0.5 mg/ml) as described previously (Luo et al, 2019;Wei G. et al, 2020). After matrine treatment, platelet aggregation induced by CRP (0.1 μg/ml) or thrombin (0.04 U/ml) was analyzed in a Lumi-Aggregometer Model 700 (Chrono-log Corporation, Havertown, PA, United States) at 37°C with stirring (1,000 rpm).…”

“…Platelet α-granule secretion was assessed via measuring P-selectin expression and integrin αIIbβ3 activation was evaluated via detecting the activation-dependent binding of PAC-1 to platelet αIIbβ3 by flow cytometry as described previously (Luo et al, 2019;Wei G. et al, 2020). Briefly, after matrine treatment, platelets were stimulated with CRP (0.1 or 2 μg/ml) or thrombin (0.04 or 0.5 U/ml) in the presence of PE-conjugated anti-P-selectin antibody or FITC-conjugated PAC-1 antibody followed by measuring P-selectin expression or PAC-1 binding by flow cytometry using FITC-conjugated mouse anti-human CD41a to set the platelet gate in the Forward Scatter and Side Scatter.…”

“…Thrombin-mediated clot retraction was initiated after addition of thrombin (1 U/ml) to washed platelets in the presence of 2 mM Ca 2+ and 0.5 mg/ml fibrinogen as described previously (Luo et al, 2019;Wei G. et al, 2020).…”

“…Mice received intraperitoneal injection of matrine (10 mg/kg). After 30 min, tail bleeding time was evaluated as described previously (Luo et al, 2019;Wei G. et al, 2020).…”

“…Deep vein thrombosis model was established through ligation of the inferior vena cava (IVC) with a 2-0 nonabsorbable suture as described previously (Wei G. et al, 2020;Wang et al, 2020). After 24 h of ligation, thrombi were excised to measure the weight and length.…”