{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T18:33:09Z","timestamp":1771007589128,"version":"3.50.1"},"reference-count":48,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2012,12,18]],"date-time":"2012-12-18T00:00:00Z","timestamp":1355788800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["British J Pharmacology"],"published-print":{"date-parts":[[2013,1]]},"abstract":"<jats:sec><jats:title>Background and Purpose<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">NADPH<\/jats:styled-content> oxidases (<jats:styled-content style=\"fixed-case\">NOXs<\/jats:styled-content>) contribute to platelet activation by a largely unknown mechanism. Here, we studied the effect of the novel <jats:styled-content style=\"fixed-case\">NOX<\/jats:styled-content> inhibitor 2\u2010acetylphenothiazine (2\u2010<jats:styled-content style=\"fixed-case\">APT<\/jats:styled-content>) on human platelet functional responses and intracellular signaling pathways.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental Approach<\/jats:title><jats:p>The generation of superoxide ions was assessed by single cell imaging on adhering platelets using dihydroethidium (<jats:styled-content style=\"fixed-case\">DHE<\/jats:styled-content>), while other reactive oxygen species (<jats:styled-content style=\"fixed-case\">ROS<\/jats:styled-content>) were detected with 5\u2010(and\u20106)\u2010carboxy\u20102\u2032,7\u2032\u2010dichlorodihydrofluorescein diacetate (<jats:styled-content style=\"fixed-case\">CM<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">H<\/jats:styled-content><jats:sub>2<\/jats:sub>\u2010<jats:styled-content style=\"fixed-case\">DCFDA<\/jats:styled-content>). Whole blood thrombus formation, washed platelet aggregation, integrin \u03b1<jats:styled-content style=\"fixed-case\">II<\/jats:styled-content>b\u03b23 inside\u2010out signalling, <jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>yk phosphorylation and <jats:styled-content style=\"fixed-case\">PKC<\/jats:styled-content> activation were analysed to understand the functional consequences of <jats:styled-content style=\"fixed-case\">NOX<\/jats:styled-content> inhibition by 2\u2010<jats:styled-content style=\"fixed-case\">APT<\/jats:styled-content> in platelets.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>Superoxide ion generation stimulated by platelet adhesion on collagen and fibrinogen was significantly inhibited by 2\u2010<jats:styled-content style=\"fixed-case\">APT<\/jats:styled-content> in concentration\u2010dependent manner (<jats:styled-content style=\"fixed-case\">IC<\/jats:styled-content><jats:sub>50<\/jats:sub> = 306\u2009n<jats:styled-content style=\"fixed-case\">M<\/jats:styled-content> and 227\u2009n<jats:styled-content style=\"fixed-case\">M<\/jats:styled-content>, respectively), whereas cumulative <jats:styled-content style=\"fixed-case\">ROS<\/jats:styled-content> accumulation was not affected by this pharmacological agent. 2\u2010<jats:styled-content style=\"fixed-case\">APT<\/jats:styled-content> also abolished collagen\u2010dependent whole blood thrombus formation and washed platelet aggregation in response to collagen but not thrombin. The activation of integrin \u03b1<jats:styled-content style=\"fixed-case\">II<\/jats:styled-content>b\u03b23 and <jats:styled-content style=\"fixed-case\">PKC<\/jats:styled-content> in response to the <jats:styled-content style=\"fixed-case\">GPVI<\/jats:styled-content>\u2010specific agonist collagen\u2010related peptide (<jats:styled-content style=\"fixed-case\">CRP<\/jats:styled-content>) was significantly reduced, whereas the same responses to thrombin were not significantly affected by 2\u2010<jats:styled-content style=\"fixed-case\">APT<\/jats:styled-content>. Finally, <jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>yk activation in response to collagen but not thrombin was inhibited by 2\u2010<jats:styled-content style=\"fixed-case\">APT<\/jats:styled-content>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Implications<\/jats:title><jats:p>Taken together, our results suggest that 2\u2010<jats:styled-content style=\"fixed-case\">APT<\/jats:styled-content> attenuates <jats:styled-content style=\"fixed-case\">GPVI<\/jats:styled-content>\u2010specific signalling and is a novel inhibitor of collagen\u2010induced platelet responses. Therefore, <jats:styled-content style=\"fixed-case\">NOXs<\/jats:styled-content> could represent a novel target for the discovery of anti\u2010thrombotic drugs.<\/jats:p><\/jats:sec>","DOI":"10.1111\/j.1476-5381.2012.02130.x","type":"journal-article","created":{"date-parts":[[2012,8,6]],"date-time":"2012-08-06T14:05:47Z","timestamp":1344261947000},"page":"212-224","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":65,"title":["The novel <scp>NOX<\/scp> inhibitor 2\u2010acetylphenothiazine impairs collagen\u2010dependent thrombus formation in a <scp>GPVI<\/scp>\u2010dependent manner"],"prefix":"10.1111","volume":"168","author":[{"given":"D","family":"Vara","sequence":"first","affiliation":[{"name":"Department of Pharmacy and Pharmacology University of Bath  Bath Spa UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M","family":"Campanella","sequence":"additional","affiliation":[{"name":"Department of Veterinary Basic Sciences, Royal Veterinary College University of London  London UK"},{"name":"Consortium for Mitochondrial Research (CfMR) University College London  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G","family":"Pula","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Pharmacology University of Bath  Bath Spa UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2012,12,18]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M701330200"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1080\/09537100701817224"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2008.03.021"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2005-03-1047"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcmd.2005.12.028"},{"key":"e_1_2_8_7_1","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1055\/s-0037-1613841","article-title":"Superoxide anion and hydroxyl radical release by collagen\u2010induced platelet aggregation \u2013 role of arachidonic acid metabolism","volume":"83","author":"Caccese D","year":"2000","journal-title":"Thromb Haemost"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0165-6147(03)00233-5"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1089\/1523086041361640"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2008-02-139733"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1161\/01.ATV.0000105889.29687.CC"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00210-010-0552-3"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrd3403"},{"key":"e_1_2_8_14_1","first-page":"21","article-title":"NADPH oxidase\u2010derived reactive oxygen species in the regulation of endothelial phenotype","volume":"60","author":"Dworakowski R","year":"2008","journal-title":"Pharmacol Rep"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.107.159178"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.00-0360fje"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2011.04192.x"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/cb100219n"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1039\/b804795b"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI108718"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2009.03722.x"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1161\/HYPERTENSIONAHA.107.100214"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M602174200"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1161\/01.ATV.18.11.1698"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0891-5849(96)00488-1"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.107.691279"},{"key":"e_1_2_8_27_1","first-page":"399","article-title":"PKCalpha regulates platelet granule secretion and thrombus formation in mice","volume":"119","author":"Konopatskaya O","year":"2009","journal-title":"J Clin Invest"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V100.3.917"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1161\/01.ATV.0000145574.90840.7d"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2002-06-1646"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.95.4.885"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1124\/jpet.111.189050"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.107.158915"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2011.04361.x"},{"key":"e_1_2_8_35_1","first-page":"3532","article-title":"Platelets and wound healing","volume":"13","author":"Nurden AT","year":"2008","journal-title":"Front Biosci"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.atherosclerosis.2004.03.021"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.0000134963.77201.55"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.05-5269com"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1080\/09537100701777303"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M409212200"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2006-05-023739"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.110.215855"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0601945103"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2008.56"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80983-6"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0049-3848(01)00341-3"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00109-010-0691-5"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2007.02.027"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1080\/09533710022149395"}],"container-title":["British Journal of Pharmacology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fj.1476-5381.2012.02130.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/j.1476-5381.2012.02130.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,30]],"date-time":"2023-10-30T04:26:56Z","timestamp":1698640016000},"score":1,"resource":{"primary":{"URL":"https:\/\/bpspubs.onlinelibrary.wiley.com\/doi\/10.1111\/j.1476-5381.2012.02130.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,12,18]]},"references-count":48,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2013,1]]}},"alternative-id":["10.1111\/j.1476-5381.2012.02130.x"],"URL":"https:\/\/doi.org\/10.1111\/j.1476-5381.2012.02130.x","archive":["Portico"],"relation":{},"ISSN":["0007-1188","1476-5381"],"issn-type":[{"value":"0007-1188","type":"print"},{"value":"1476-5381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,12,18]]},"assertion":[{"value":"2012-02-22","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2012-07-20","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2012-12-18","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}