Platelet hyperactivity is essential for thrombus formation in coronary artery diseases(CAD).Dysfunction of the cystic fibrosis transmembrane conductance regulator(CFTR)in patients with cystic fibrosis elevates intracellular Cl-levels([Cl-]i)and enhanced platelet hyperactivity.In this study,we explored whether alteration of[Cl-]i has a pathological role in regulating platelet hyperactivity and arterial thrombosis formation.CFTR expression was significantly decreased,while[Cl-]i was increased in platelets from CAD patients.In a FeCl3-induced mouse mesenteric arteriole thrombosis model,platelet-specific Cftr-knockout and/or pre-administration of ion channel inhibitor CFTRinh-172 increased platelet[Cl-]i,which accelerated thrombus formation,enhanced platelet aggregation and ATP release,and increased P2Y12 and PAR4 expression in platelets.Conversely,Cftr-overexpressing platelets resulted in subnormal[Cl-]i,thereby decreasing thrombosis formation.Our results showed that clamping[Cl-]i at high levels or Cftr deficiency-induced[Cl-]i increasement dramatically augmented phosphorylation(Ser422)of serum and glucocorticoid-regulated kinase(SGK1),subsequently upregulated P2Y12 and PAR4 expression via NF-KB signaling.Constitutively active mutant S422D SGK1 markedly increased P2Y12 and PAR4 expression.The specific SGK1 inhibitor GSK-650394 decreased platelet aggregation in wildtype and platelet-specific Cftr knockout mice,and platelet SGK1 phosphorylation was observed in line with increased[Cl-]i and decreased CFTR expression in CAD patients.Co-transfection of S422D SGK1 and adenovirus-induced CFTR overexpression in MEG-01 cells restored platelet activation signaling cascade.Our results suggest that[Cl-]i is a novel positive regulator of platelet activation and arterial thrombus formation via the activation of a[Cl-]i-sensitive SGK1 signaling pathway.Therefore,[Cl-]i in platelets is a novel potential biomarker for platelet hyperactivity,and CFTR may be a potential therapeutic target for platelet activation in CAD.

Han-yan Yang;Chao Zhang;Liang Hu;Chang Liu;Ni Pan;Mei Li;Hui Han;Yi Zhou;Jie Li;Li-yan Zhao;Yao-sheng Liu;Bing-zheng Luo;Xiong-qing Huang;Xiao-fei Lv;Zi-cheng Li;Jun Li;Zhi-hong Li;Ruo-mei Wang;Li Wang;Yong-yuan Guan;Can-zhao Liu;Bin Zhang;Guan-lei Wang

Department of Pharmacology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510080,China;Academy of Integrative Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;institute of Pediatrics,Guangzhou Women and Children's Medical Center affiliated to Guangzhou Medical College,Guangzhou 510623,China;VIP Healthcare Center,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China;Guangdong Cardiovascular Institute,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China;Department of Anesthesiology,The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,China;Department of Pharmacy,The First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China;Department of Anesthesiology,the First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China;School of Computer Science and Engineering,Sun Yat-sen University,Guangzhou 510006,China;Division of Biological Sciences,University of California,San Diego,La Jolla,CA,USA;Department of Cardiovascular Medicine,Translational Medicine Research Center,Zhujiang Hospital,Southern Medical University,Guangzhou 510280,China

中国药理学报（英文版）

[1]Han-yan Yang;Chao Zhang;Liang Hu;Chang Liu;Ni Pan;Mei Li;Hui Han;Yi Zhou;Jie Li;Li-yan Zhao;Yao-sheng Liu;Bing-zheng Luo;Xiong-qing Huang;Xiao-fei Lv;Zi-cheng Li;Jun Li;Zhi-hong Li;Ruo-mei Wang;Li Wang;Yong-yuan Guan;Can-zhao Liu;Bin Zhang;Guan-lei Wang-.Platelet CFTR inhibition enhances arterial thrombosis via increasing intracellular Cl-concentration and activation of SGK1 signaling pathway)[J].中国药理学报（英文版）,2022(10):2596-2608

Cftr,CFTRinh,subnormal,Ser422,Constitutively,S422D

Platelet,inhibition,enhances,arterial,thrombosis,via,increasing,intracellular,concentration,activation,SGK1,signaling,pathway,hyperactivity,essential,thrombus,formation,coronary,artery,diseases,CAD,Dysfunction,cystic,fibrosis,transmembrane,conductance,regulator,patients,elevates,levels,enhanced,In,this,study,explored,whether,alteration,has,pathological,role,regulating,was,significantly,decreased,while,increased,platelets,from,FeCl3,induced,mouse,mesenteric,arteriole,model,specific,knockout,administration,channel,inhibitor,which,accelerated,aggregation,ATP,release,P2Y12,PAR4,Conversely,overexpressing,resulted,thereby,decreasing,Our,results,showed,that,clamping,high,deficiency,increasement,dramatically,augmented,phosphorylation,serum,glucocorticoid,kinase,subsequently,upregulated,KB,active,mutant,markedly,GSK,wildtype,mice,observed,line,transfection,adenovirus,overexpression,MEG,cells,restored,cascade,suggest,novel,positive,sensitive,Therefore,potential,biomarker,may,be,therapeutic,target

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