Colchicine inhibits ROS generation in response to glycoprotein VI stimulation

G J Pennings,V Chen,C J Reddel,M Traini,L Kritharides,H Campbell

doi:10.1038/s41598-021-91409-7

Abstract

Colchicine inhibits coronary and cerebrovascular events in patients with coronary artery disease (CAD), and although known to have anti-inflammatory properties, its mechanisms of action are incompletely understood. In this study, we investigated the effects of colchicine on platelet activation with a particular focus on its effects on activation via the collagen glycoprotein (GP)VI receptor, P2Y12 receptor, and procoagulant platelet formation. Therapeutic concentrations of colchicine in vitro (equivalent to plasma levels) significantly decreased platelet aggregation in whole blood and in platelet rich plasma in response to collagen (multiplate aggregometry) and reduced reactive oxygen species (ROS) generation (H2DCF-DA, flow cytometry) in response to GPVI stimulation with collagen related peptide-XL (CRP-XL, GPVI specific agonist). Other platelet activation pathways including P-selectin expression, GPIIb/IIIa conformational change and procoagulant platelet formation (GSAO+/CD62P+) (flow cytometry) were inhibited with higher concentrations of colchicine known to inhibit microtubule depolymerization. Pathway specific mechanisms of action of colchicine on platelets, including modulation of the GPVI receptor pathway at low concentrations, may contribute to its protective role in CAD.

Highlights

Colchicine is well known as an anti-inflammatory drug used in the treatment of acute gouty arthritis, familial Mediterranean fever and Behçet’s disease[1]
We investigated the effect of colchicine, at therapeutic plasma concentration and at microtubule depolymerisation concentration, on platelet aggregation in response to collagen and adenosine diphosphate (ADP), and the downstream effects of colchicine after GPVI and P 2Y12 stimulation on reactive oxygen species (ROS) generation and calcium flux, respectively
In this study we investigated the effects of circulating therapeutic concentrations (20 nM) and microtubule depolymerisation-inhibitory concentrations (2 mM) of colchicine on platelet activation

Summary

Introduction

Colchicine is well known as an anti-inflammatory drug used in the treatment of acute gouty arthritis, familial Mediterranean fever and Behçet’s disease[1] It has been used for the treatment of pericarditis[2], atrial fibrillation post-coronary bypass, stroke and for secondary prevention of cardiac e vents[3,4,5,6,7]. Studies demonstrated a colchicine concentration-dependent decrease in aggregation in response to ADP, epinephrine and collagen in vitro[21,22,23,24,25,26]. Recent studies have identified that anti-GPVI therapy can lead to disaggregation of thrombus in the absence of t hrombin[41]

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Results

Conclusion