Cargando…

Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response

BACKGROUND: Deep 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...

Descripción completa

Detalles Bibliográficos
Autores principales: Chen, Ziyu, Wang, Gaorui, Xie, Xueqing, Liu, Heng, Liao, Jun, Shi, Hailian, Chen, Min, Lai, Shusheng, Wang, Zhengtao, Wu, Xiaojun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9411522/
https://www.ncbi.nlm.nih.gov/pubmed/36032109
http://dx.doi.org/10.3389/fimmu.2022.918476
_version_ 1784775287542644736
author Chen, Ziyu
Wang, Gaorui
Xie, Xueqing
Liu, Heng
Liao, Jun
Shi, Hailian
Chen, Min
Lai, Shusheng
Wang, Zhengtao
Wu, Xiaojun
author_facet Chen, Ziyu
Wang, Gaorui
Xie, Xueqing
Liu, Heng
Liao, Jun
Shi, Hailian
Chen, Min
Lai, Shusheng
Wang, Zhengtao
Wu, Xiaojun
author_sort Chen, Ziyu
collection PubMed
description BACKGROUND: Deep 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. METHODS: The interaction between Rg5 and P2RY(12) 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 P2RY(12). RESULTS: Rg5 allosterically interacted with P2RY(12), 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 Ca(2+) concentration, thus reducing the activity and expression of peptidyl arginine deiminase 4 to prevent NETosis. The inhibitory effect on neutrophil activity was dependent on P2RY(12). CONCLUSIONS: Rg5 could attenuate experimental DVT by counteracting NETosis and inflammatory response in neutrophils via P2RY(12), which may pave the road for its clinical application in the prevention of DVT-related disorders.
format Online
Article
Text
id pubmed-9411522
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-94115222022-08-27 Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response Chen, Ziyu Wang, Gaorui Xie, Xueqing Liu, Heng Liao, Jun Shi, Hailian Chen, Min Lai, Shusheng Wang, Zhengtao Wu, Xiaojun Front Immunol Immunology BACKGROUND: Deep 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. METHODS: The interaction between Rg5 and P2RY(12) 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 P2RY(12). RESULTS: Rg5 allosterically interacted with P2RY(12), 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 Ca(2+) concentration, thus reducing the activity and expression of peptidyl arginine deiminase 4 to prevent NETosis. The inhibitory effect on neutrophil activity was dependent on P2RY(12). CONCLUSIONS: Rg5 could attenuate experimental DVT by counteracting NETosis and inflammatory response in neutrophils via P2RY(12), which may pave the road for its clinical application in the prevention of DVT-related disorders. Frontiers Media S.A. 2022-08-12 /pmc/articles/PMC9411522/ /pubmed/36032109 http://dx.doi.org/10.3389/fimmu.2022.918476 Text en Copyright © 2022 Chen, Wang, Xie, Liu, Liao, Shi, Chen, Lai, Wang and Wu https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Chen, Ziyu
Wang, Gaorui
Xie, Xueqing
Liu, Heng
Liao, Jun
Shi, Hailian
Chen, Min
Lai, Shusheng
Wang, Zhengtao
Wu, Xiaojun
Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response
title Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response
title_full Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response
title_fullStr Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response
title_full_unstemmed Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response
title_short Ginsenoside Rg5 allosterically interacts with P2RY(12) and ameliorates deep venous thrombosis by counteracting neutrophil NETosis and inflammatory response
title_sort ginsenoside rg5 allosterically interacts with p2ry(12) and ameliorates deep venous thrombosis by counteracting neutrophil netosis and inflammatory response
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9411522/
https://www.ncbi.nlm.nih.gov/pubmed/36032109
http://dx.doi.org/10.3389/fimmu.2022.918476
work_keys_str_mv AT chenziyu ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT wanggaorui ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT xiexueqing ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT liuheng ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT liaojun ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT shihailian ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT chenmin ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT laishusheng ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT wangzhengtao ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse
AT wuxiaojun ginsenosiderg5allostericallyinteractswithp2ry12andamelioratesdeepvenousthrombosisbycounteractingneutrophilnetosisandinflammatoryresponse