Ginsenoside Rg5 allosterically interacts with P2RY12 and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response

Abstract

BackgroundDeep venous thrombosis (DVT) highly occurs in patients with severe COVID-19 and probably accounted for their high mortality. DVT formation is a time-dependent inflammatory process in which NETosis plays an important role. However, whether ginsenoside Rg5 from species of Panax genus could alleviate DVT and its underlying mechanism has not been elucidated.MethodsThe interaction between Rg5 and P2RY12 was studied by molecular docking, molecular dynamics, surface plasmon resonance (SPR), and molecular biology assays. The preventive effect of Rg5 on DVT was evaluated in inferior vena cava stasis–induced mice, and immunocytochemistry, Western blot, and calcium flux assay were performed in neutrophils from bone marrow to explore the mechanism of Rg5 in NETosis via P2RY12.ResultsRg5 allosterically interacted with P2RY12, formed stable complex, and antagonized its activity via residue E188 and R265. Rg5 ameliorated the formation of thrombus in DVT mice; accompanied by decreased release of Interleukin (IL)-6, IL-1β, and tumor necrosis factor-α in plasma; and suppressed neutrophil infiltration and neutrophil extracellular trap (NET) release. In lipopolysaccharide- and platelet-activating factor–induced neutrophils, Rg5 reduced inflammatory responses via inhibiting the activation of ERK/NF-κB signaling pathway while decreasing cellular Ca2+ concentration, thus reducing the activity and expression of peptidyl arginine deiminase 4 to prevent NETosis. The inhibitory effect on neutrophil activity was dependent on P2RY12.ConclusionsRg5 could attenuate experimental DVT by counteracting NETosis and inflammatory response in neutrophils via P2RY12, which may pave the road for its clinical application in the prevention of DVT-related disorders.

NET-targeted nanoparticles for antithrombotic therapy in pregnancy

“…Controlled sustained release and targeted release of drugs in drug delivery systems can be triggered by many physical and chemical stimuli in the microenvironment, such as light, heat, magnetism, and pH. 28 , 29 Under laser irradiation, the in vitro release rate of PB NPs in PB-NEBP liposomes was significantly higher than that in the nonirradiated group at same time point ( Figure 2 D). This may be because after laser irradiation, the PB-NEBP liposomes generate heat, which accelerates the degradation rate of the polymer and accelerates the release of PB NPs.…”

“…One of the main ways that NETs promote thrombosis is to provide a scaffold for the adhesion of platelets and red blood cells. 29 , 30 The formation of this stent is not only the structural basis for thrombosis, but many components on the stent can also actively activate the coagulation process. Neutrophils release NETs after activation.…”

Identification of potential immunotherapeutic targets and prognostic biomarkers in Graves' disease using weighted gene co-expression network analysis

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Neutrophil extracellular traps mediate deep vein thrombosis: from mechanism to therapy

“…Ginsenoside Rg5, a natural saponin in P. ginseng and P. notoginseng, interacts with the P2RY12 variant to decrease the inflammatory response by inhibiting the ERK/NF-κb signaling pathway, suppressing neutrophil infiltration and NET release, and interfering with DVT formation in mice. This was accompanied by a reduction in plasma IL-6, IL-1β, and TNF-α release, which may have clinical implications for the prevention of DVT-related clinical disorders ( 224 ). These findings provide new research directions for clinical antithrombotic therapy.…”