Inhibition of Shear-Induced Platelet Aggregation by Xueshuantong via Targeting Piezo1 Channel-Mediated Ca2+ Signaling Pathway

Abstract

XueShuanTong (XST) comprising therapeutically active ginsenosides, a lyophilized extract of Panax notoginseng roots, is extensively used in traditional Chinese medicine to treat ischemic heart and cerebrovascular diseases. Our recent study shows that treatment with XST inhibits shear-induced thrombosis formation but the underlying mechanism remained unclear. This study aimed to investigate the hypothesis that XST inhibited shear-induced platelet aggregation via targeting the mechanosensitive Ca2+-permeable Piezo1 channel by performing platelet aggregation assay, Ca2+ imaging and Western blotting analysis. Exposure to shear at physiologically (1,000–2000 s−1) and pathologically related rates (4,000–6,000 s−1) induced platelet aggregation that was inhibited by treatment with GsMTx-4. Exposure to shear evoked robust Ca2+ responses in platelets that were inhibited by treatment with GsMTx-4 and conversely enhanced by treatment with Yoda1. Treatment with XST at a clinically relevant concentration (0.15 g L−1) potently inhibited shear-induced Ca2+ responses and platelet aggregation, without altering vWF-mediated platelet adhesion and rolling. Exposure to shear, while resulting in no effect on the calpain-2 expression in platelets, induced calpain-2-mediated cleavage of talin1 protein, which is known to be critical for platelet activation. Shear-induced activation of calpain-2 and cleavage of talin1 were attenuated by treatment with XST. Taken together, our results suggest that XST inhibits shear-induced platelet aggregation via targeting the Piezo1 channel to prevent Piezo1-mediated Ca2+ signaling and downstream calpain-2 and talin1 signal pathway, thus providing novel insights into the mechanism of the therapeutic action of XST on platelet aggregation and thrombosis formation.

Ginseng in vascular dysfunction: A review of therapeutic potentials and molecular mechanisms

“…Ginsenoside Rk1 inhibited platelet‐mediated thrombus formation by downregulation of granule release and αβ activation (J. H. Shin, Kwon, Irfan, Rhee, & Lee, 2021). Xueshuantong is composed of ginsenosides to treat ischemic heart and cerebrovascular diseases, and it was found that Xueshuantong suppressed shear‐induced platelet aggregation by targeting piezo1 channel‐mediated Ca 2+ signaling (L. Liu et al, 2021). Meanwhile, another article reported that red GE was embedded in nanomaterials or antithrombotic materials to improve the oral availability of red ginseng and its antithrombotic activity (E. S. Kim, Lee, & Lee, 2021).…”

Piezo Channels: Awesome Mechanosensitive Structures in Cellular Mechanotransduction and Their Role in Bone

“…When Piezo channels are opened/activated, a cellular signaling cascade is triggered by the Ca 2+ influx through the Piezo channels [ 24 , 50 , 51 , 52 , 53 ]. Zhao et al considered that the extracellular CED has a lot of negative charge, so extracellular cations such as Ca 2+ will be aggregated and will flow through the pore of Piezo channels to cross the cell membrane [ 24 ].…”

“…By experimentally stretching epithelial cells in vitro, Gudipaty et al found that mechanical stretch, through the mechanosensitive Piezo1, triggers these cells to pause in early G2 to activate calcium-dependent ERK1/2 phosphorylation and cyclin B transcription, thus prompting these cells to enter mitosis [ 52 ]. Exposure to blood flow shear force in vitro activates the Piezo1 channel, with ensuing Piezo1-mediated Ca 2+ influx in platelets [ 53 ]. An increase in intracellular Ca 2+ level in turn promotes activation of calpain-2 and cleavage of talin1, leading to platelet aggregation [ 53 ].…”

“…Exposure to blood flow shear force in vitro activates the Piezo1 channel, with ensuing Piezo1-mediated Ca 2+ influx in platelets [ 53 ]. An increase in intracellular Ca 2+ level in turn promotes activation of calpain-2 and cleavage of talin1, leading to platelet aggregation [ 53 ]. The signaling cascade induced by Ca 2+ flowing through Piezo channels also takes place in bone cells and chondrocytes [ 7 , 26 , 32 ].…”

Platelet Mechanobiology Inspired Microdevices: From Hematological Function Tests to Disease and Drug Screening

“…At the molecular scale, increasing evidences suggest that platelets can undergo mechanosensing upon receiving these hemodynamic stimuli. Key players in such mechanosensing processes include von Willebrand factors (VWF) ( Savage et al, 1996 ; Fu et al, 2017 ), fibrinogen ( Butera and Hogg, 2020 ; Peshkova et al, 2020 ), mechanoreceptors including glycoprotein Ib (GPIb) ( Chen et al, 2016 ; Ju et al, 2016 ) and glycoprotein IIb/IIIa (GPIIb/IIIa or integrin α IIb β 3 ) ( Nesbitt et al, 2009 ; Chen Y et al, 2019 ), and mechanosensitive ion channels ( Abbonante et al, 2017 ; Ilkan et al, 2017 ; Liu et al, 2021 ). Further, activated platelets generate contractile forces to stabilize and consolidate the thrombus ( Osdoit and Rosa, 2001 ; Ono et al, 2008 ; Hansen et al, 2018 ).…”