{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T22:25:42Z","timestamp":1791584742658,"version":"4.3.4"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,9,28]],"date-time":"2023-09-28T00:00:00Z","timestamp":1695859200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Excessive neutrophil extracellular traps (NETs) is involved in the progression of acute pancreatitis (AP) but the mechanisms controlling NETs formation in AP are not fully understood. Therefore, our study sought to investigate the mechanism of the highly expressed P-selectin stimulating the formation of NETs in AP.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>NETs formation was detected by flow cytometry, immunofluorescence staining, and cf-DNA and MPO-DNA complexes were measured as biomarkers of NETs formation. Neutrophils treated with P-selectin and pharmacological inhibitors were examined by western blot, immunofluorescence staining and flow cytometry. Mouse model of AP was established by caerulein and the effect of inhibiting P-selectin by PSI-697 on the level of NETs and PAD4 in pancreatic tissue was observed. The severity of AP was evaluated by histopathological score and the detection of serum amylase and lipase.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Patients with AP had elevated levels of NETs and P-selectin compared with healthy volunteers. Stimulation of P-selectin up-regulated the expression of PSGL-1, increased the phosphorylation of Syk, mediated intracellular calcium signal and led to the activation and expression of PAD4, which modulated NETs formation in neutrophils. Pretreament with PSI-697 blunted NETs formation and PAD4 expression in the pancreatic tissue, and ameliorated the severity of AP in mice.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Taken together, these results suggest that P-selectin induces NETs through PSGL-1 and its downstream Syk\/Ca<jats:sup>2+<\/jats:sup>\/PAD4 signaling pathway, and that targeting this pathway might be a promising strategy for the treatment of AP.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2023.1265344","type":"journal-article","created":{"date-parts":[[2023,9,29]],"date-time":"2023-09-29T03:59:10Z","timestamp":1695959950000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":31,"title":["High expression of P-selectin induces neutrophil extracellular traps via the PSGL-1\/Syk\/Ca2+\/PAD4 pathway to exacerbate acute pancreatitis"],"prefix":"10.3389","volume":"14","author":[{"given":"Qi","family":"Xu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ming","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Xia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang","family":"Luo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaofang","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoying","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David Y. B.","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,9,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"e1","DOI":"10.1016\/j.pan.2013.07.063","article-title":"IAP\/APA evidence-based guidelines for the management of acute pancreatitis","volume":"13","author":"Besselink","year":"2013","journal-title":"Pancreatology"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(20)31310-6","article-title":"Acute pancreatitis","volume":"396","author":"Boxhoorn","year":"2020","journal-title":"Lancet (London England)"},{"key":"B3","article-title":"Pathophysiology of acute pancreatitis","volume":"40","author":"Glasbrenner","year":"1993","journal-title":"Hepato-gastroenterology"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/j.pan.2013.01.010","article-title":"Patients with acute pancreatitis complicated by organ dysfunction show abnormal peripheral blood polymorphonuclear leukocyte signaling","volume":"13","author":"Oiva","year":"2013","journal-title":"Pancreatology"},{"key":"B5","doi-asserted-by":"publisher","first-page":"320803519","DOI":"10.1177\/1756284820974913","article-title":"Targeting neutrophil extracellular traps in severe acute pancreatitis treatment","volume":"13","author":"Hu","year":"2020","journal-title":"Therap Adv Gastroenterol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1126\/science.1092385","article-title":"Neutrophil extracellular traps kill bacteria","volume":"303","author":"Brinkmann","year":"2004","journal-title":"Sci (New York N.Y.)"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2015.08.026","article-title":"Neutrophil extracellular traps induce trypsin activation, inflammation, and tissue damage in mice with severe acute pancreatitis","volume":"149","author":"Merza","year":"2015","journal-title":"Gastroenterology"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1038\/s41374-020-0469-5","article-title":"Extracellular cold-inducible RNA-binding protein regulates neutrophil extracellular trap formation and tissue damage in acute pancreatitis","volume":"100","author":"Linders","year":"2020","journal-title":"Lab investigation; J Tech Methods Pathol"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms10973","article-title":"Externalized decondensed neutrophil chromatin occludes pancreatic ducts and drives pancreatitis","volume":"7","author":"Leppkes","year":"2016","journal-title":"Nat Commun"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1002\/bjs.7775","article-title":"P-selectin mediates neutrophil rolling and recruitment in acute pancreatitis","volume":"99","author":"Hartman","year":"2012","journal-title":"Br J Surg"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1007\/s002100000268","article-title":"Critical role of P-selectin-dependent rolling in tumor necrosis factor-alpha-induced leukocyte adhesion and extravascular recruitment in vivo","volume":"362","author":"M\u00e5nsson","year":"2000","journal-title":"Naunyn-Schmiedeberg's Arch Pharmacol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00435.2004","article-title":"Treatment with bindarit, a blocker of MCP-1 synthesis, protects mice against acute pancreatitis","volume":"288","author":"Bhatia","year":"2005","journal-title":"Am J Physiol Gastrointestinal liver Physiol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2012.12.042","article-title":"Role of immune cells and immune-based therapies in pancreatitis and pancreatic ductal adenocarcinoma","volume":"144","author":"Zheng","year":"2013","journal-title":"Gastroenterology"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1111\/jcmm.12639","article-title":"Central role of neutrophil in the pathogenesis of severe acute pancreatitis","volume":"19","author":"Yang","year":"2015","journal-title":"J Cell Mol Med"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1016\/j.thromres.2019.11.028","article-title":"Microparticles from aged packed red blood cell units stimulate pulmonary microthrombus formation via P-selectin","volume":"185","author":"Kim","year":"2020","journal-title":"Thromb Res"},{"key":"B16","doi-asserted-by":"publisher","first-page":"899","DOI":"10.1517\/14728222.2010.504717","article-title":"Targeting P-selectin in acute pancreatitis","volume":"14","author":"Hackert","year":"2010","journal-title":"Expert Opin Ther Targets"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1097\/00000658-200002000-00010","article-title":"Quantitative measurement of P- and E-selectin adhesion molecules in acute pancreatitis: correlation with distant organ injury","volume":"231","author":"Lundberg","year":"2000","journal-title":"Ann Surg"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1021\/acsbiomaterials.0c00596","article-title":"P-selectin-based dual-model nanoprobe used for the specific and rapid visualization of early detection toward severe acute pancreatitis in vivo","volume":"6","author":"Long","year":"2020","journal-title":"ACS Biomater Sci Eng"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1097\/MPA.0000000000001009","article-title":"E and P selectins as potential markers in the assessment of the severity of acute pancreatitis","volume":"47","author":"Tsaroucha","year":"2018","journal-title":"Pancreas"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2015-01-624023","article-title":"P-selectin promotes neutrophil extracellular trap formation in mice","volume":"126","author":"Etulain","year":"2015","journal-title":"Blood"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.24185","article-title":"Flow cytometric assay for direct quantification of neutrophil extracellular traps in blood samples","volume":"90","author":"Gavillet","year":"2015","journal-title":"Am J Hematol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0148142","article-title":"Neutrophil extracellular traps induce organ damage during experimental and clinical sepsis","volume":"11","author":"Czaikoski","year":"2016","journal-title":"PloS One"},{"key":"B23","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1097\/00000658-199201000-00007","article-title":"A better model of acute pancreatitis for evaluating therapy","volume":"215","author":"Schmidt","year":"1992","journal-title":"Ann Surg"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2016-313595","article-title":"Acute pancreatitis: recent advances through randomised trials","volume":"66","author":"van Dijk","year":"2017","journal-title":"Gut"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.974821","article-title":"Roles, detection, and visualization of neutrophil extracellular traps in acute pancreatitis","volume":"13","author":"Li","year":"2022","journal-title":"Front Immunol"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1002\/JLB.3A0322-151R","article-title":"Epigallocatechin-3-gallate reduces neutrophil extracellular trap formation and tissue injury in severe acute pancreatitis","volume":"112","author":"Li","year":"2022","journal-title":"J Leukoc Biol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2022.927193","article-title":"Damage associated molecular patterns and neutrophil extracellular traps in acute pancreatitis","volume":"12","author":"Zhou","year":"2022","journal-title":"Front Cell Infect Microbiol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1002\/jlb.59.2.229","article-title":"Rapid killing of human neutrophils by the potent activator phorbol 12-myristate 13-acetate (PMA) accompanied by changes different from typical apoptosis or necrosis","volume":"59","author":"Takei","year":"1996","journal-title":"J Leukoc Biol"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1038\/nm1565","article-title":"Data from: Platelet TLR4 activates neutrophil extracellular traps to ensnare bacteria in septic blood","volume":"13","author":"Clark","year":"2007","journal-title":"Nat Med"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2014-308043","article-title":"Peripancreatic fat necrosis worsens acute pancreatitis independent of pancreatic necrosis via unsaturated fatty acids increased in human pancreatic necrosis collections","volume":"65","author":"Noel","year":"2016","journal-title":"Gut"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1038\/nri.2017.105","article-title":"Neutrophil extracellular traps in immunity and disease","volume":"18","author":"Papayannopoulos","year":"2018","journal-title":"Nat Rev Immunol"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2847","article-title":"Infection-induced NETosis is a dynamic process involving neutrophil multitasking in vivo","volume":"18","author":"Yipp","year":"2012","journal-title":"Nat Med"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.24437","article-title":"Diverse stimuli engage different neutrophil extracellular trap pathways","volume":"6","author":"Kenny","year":"2017","journal-title":"Elife"},{"key":"B34","doi-asserted-by":"publisher","first-page":"3409","DOI":"10.1038\/s41598-017-03257-z","article-title":"JNK activation turns on LPS- and gram-negative bacteria-induced NADPH oxidase-dependent suicidal NETosis","volume":"7","author":"Khan","year":"2017","journal-title":"Sci Rep"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2012.00307","article-title":"PAD4 mediated histone hypercitrullination induces heterochromatin decondensation and chromatin unfolding to form neutrophil extracellular trap-like structures","volume":"3","author":"Leshner","year":"2012","journal-title":"Front Immunol"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1200419109","article-title":"Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-associated thrombosis","volume":"109","author":"Demers","year":"2012","journal-title":"Proc Natl Acad Sci U S"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20100239","article-title":"PAD4 is essential for antibacterial innate immunity mediated by neutrophil extracellular traps","volume":"207","author":"Li","year":"2010","journal-title":"J Exp Med"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-019-03237-8","article-title":"An interplay of structure and intrinsic disorder in the functionality of peptidylarginine deiminases, a family of key autoimmunity-related enzymes","volume":"76","author":"Alghamdi","year":"2019","journal-title":"Cell Mol Life Sci CMLS"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1016\/0009-9120(90)80030-M","article-title":"The calcium signal and neutrophil activation","volume":"23","author":"Krause","year":"1990","journal-title":"Clin Biochem"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2004.11.152","article-title":"Comparison of enzymatic properties between hPADI2 and hPADI4","volume":"327","author":"Nakayama-Hamada","year":"2005","journal-title":"Biochem Biophys Res Commun"},{"key":"B41","doi-asserted-by":"publisher","first-page":"2429","DOI":"10.1038\/s41598-017-02677-1","article-title":"Probing the roles of calcium-binding sites during the folding of human peptidylarginine deiminase 4","volume":"7","author":"Liu","year":"2017","journal-title":"Sci Rep"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1007\/s13238-016-0364-4","article-title":"Force-dependent calcium signaling and its pathway of human neutrophils on P-selectin in flow","volume":"8","author":"Huang","year":"2017","journal-title":"Protein Cell"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(02)00420-X","article-title":"ITAM-based interaction of ERM proteins with Syk mediates signaling by the leukocyte adhesion receptor PSGL-1","volume":"17","author":"Urzainqui","year":"2002","journal-title":"Immunity"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1111\/eci.12964","article-title":"Calcium signalling and related ion channels in neutrophil recruitment and function","author":"Immler","year":"2018","journal-title":"Eur J Clin Invest"},{"key":"B45","doi-asserted-by":"publisher","first-page":"1127","DOI":"10.3390\/biomedicines9091127","article-title":"The fc\u03b3RIII engagement augments PMA-stimulated neutrophil extracellular traps (NETs) formation by granulocytes partially via cross-talk between syk-ERK-NF-\u03baB and PKC-ROS signaling pathways","volume":"9","author":"Lu","year":"2021","journal-title":"Biomedicines"},{"key":"B46","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1159\/000365249","article-title":"Src family kinases and Syk are required for neutrophil extracellular trap formation in response to \u03b2-glucan particles","volume":"7","author":"Nan\u00ec","year":"2015","journal-title":"J Innate Immun"},{"key":"B47","doi-asserted-by":"publisher","first-page":"e1008096","DOI":"10.1371\/journal.ppat.1008096","article-title":"Candida albicans triggers NADPH oxidase-independent neutrophil extracellular traps through dectin-2","volume":"15","author":"Wu","year":"2019","journal-title":"PloS Pathog"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1124\/jpet.107.128124","article-title":"Characterization of the novel P-selectin inhibitor PSI-697 [2-(4-chlorobenzyl)-3-hydroxy-7,8,9,10-tetrahydrobenzo[h] quinoline-4-carboxylic acid] in vitro and in rodent models of vascular inflammation and thrombosis","volume":"324","author":"Bedard","year":"2008","journal-title":"J Pharmacol Exp Ther"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1160\/TH06-11-0658","article-title":"Resolution of venous thrombosis using a novel oral small-molecule inhibitor of P-selectin (PSI-697) without anticoagulation","volume":"97","author":"Myers","year":"2007","journal-title":"Thromb Haemost"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2009.03587.x","article-title":"P-selectin antagonism reduces thrombus formation in humans","volume":"7","author":"Chelliah","year":"2009","journal-title":"J Thromb haemostasis JTH"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1161\/JAHA.112.006007","article-title":"Effect of PSI-697, a novel P-selectin inhibitor, on platelet-monocyte aggregate formation in humans","volume":"2","author":"Japp","year":"2013","journal-title":"J Am Heart Assoc"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1265344\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,29]],"date-time":"2023-09-29T03:59:14Z","timestamp":1695959954000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1265344\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,28]]},"references-count":51,"alternative-id":["10.3389\/fimmu.2023.1265344"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2023.1265344","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,28]]},"article-number":"1265344"}}