Rutaecarpine, an Alkaloid from Evodia rutaecarpa, Can Prevent Platelet Activation in Humans and Reduce Microvascular Thrombosis in Mice: Crucial Role of the PI3K/Akt/GSK3β Signal Axis through a Cyclic Nucleotides/VASP-Independent Mechanism.

Chun-Jen Huang,Periyakali Saravana Bhavan,Chih-Wei Hsia,Cheng-Ying Hsieh,Oanh-Thi Tran,Lan-Hsin Shu,Wei-Chieh Huang,Thanasekaran Jayakumar,Chao-Chien Chang,Wei-Ting Lin,Joen-Rong Sheu

doi:10.3390/ijms222011109

Chun-Jen Huang, Periyakali Saravana Bhavan + Show 9 more

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https://doi.org/10.3390/ijms222011109

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Abstract

The role of activated platelets in acute and chronic cardiovascular diseases (CVDs) is well established. Therefore, antiplatelet drugs significantly reduce the risk of severe CVDs. Evodia rutaecarpa (Wu-Chu-Yu) is a well-known Chinese medicine, and rutaecarpine (Rut) is a main bioactive component with substantial beneficial properties including vasodilation. To address a research gap, we investigated the inhibitory mechanisms of Rut in washed human platelets and experimental mice. At low concentrations (1–5 μM), Rut strongly inhibited collagen-induced platelet aggregation, whereas it exerted only a slight or no effect on platelets stimulated with other agonists (e.g., thrombin). Rut markedly inhibited P-selectin expression; adenosine triphosphate release; [Ca2+]i mobilization; hydroxyl radical formation; and phospholipase C (PLC)γ2/protein kinase C (PKC), mitogen-activated protein kinase, and phosphoinositide 3-kinase (PI3K)/Akt/glycogen synthase kinase-3β (GSK3β) phosphorylation stimulated by collagen. SQ22536 (an adenylate cyclase inhibitor) or ODQ (a guanylate cyclase inhibitor) did not reverse Rut-mediated antiplatelet aggregation. Rut was not directly responding to vasodilator-stimulated phosphoprotein phosphorylation. Rut significantly increased the occlusion time of fluorescence irradiated thrombotic platelet plug formation. The findings demonstrated that Rut exerts a strong effect against platelet activation through the PLCγ2/PKC and PI3K/Akt/GSK3β pathways. Thus, Rut can be a potential therapeutic agent for thromboembolic disorders.

Highlights

Traditional Chinese herbs have long been used as vital remedies worldwide
The results of the current study revealed that Rut (1–5 μM) most strongly inhibited platelet aggregation stimulated by collagen (1 μg/mL) in washed human platelets (Figure 1A,E); it exerted only a slight or no effect on platelets stimulated with either thrombin (0.02 U/mL), arachidonic acid (AA; 60 μM), or U46619 (1 μM), a prostaglandin endoperoxide, or ADP (20 μM), even at concentrations up to 100 μM (Figure 1B–E)
At concentrations of 2.5 and 5 μM, Rut markedly inhibited the phosphorylation of the phosphoinositide 3-kinase (PI3K)/Akt/glycogen synthase kinase-3β (GSK3β) pathway under collagen stimulation (Figure 5). These results indicate that the inhibition of Mitogen-activated protein kinase (MAPK) and the PI3K/Akt/GSK3β pathway may contribute critically to antiplatelet effects of Rut

Summary

Introduction

Traditional Chinese herbs have long been used as vital remedies worldwide. Rutaecarpine (Rut) aceous plants, Evodia rutaecarpa (the dried fruit of this plant is known as “Wu-Chu-Yu” in China), have long been in traditional Chinese medication for the treatment of gastrointestinal disorders, headache, amenorrhea, and postpartum hemorrhage [1]. Several alkaloids—including three major alkaloids, namely dehydroevodiamine, evodiamine, and Rut—have been identified in E. rutaecarpa and exhibit potent pharmacological activities. Yamahara et al [4] reported that Rut exerted a potent antianoxic effect on a potassium cyanide-induced cerebral anoxia model. These findings indicate that Rut can have beneficial effects on cardiovascular activities

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