Endocannabinoids Control Platelet Activation and Limit Aggregate Formation under Flow
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{"title"=>"Endocannabinoids control platelet activation and limit aggregate formation under flow", "type"=>"journal", "authors"=>[{"first_name"=>"Valentina", "last_name"=>"De Angelis", "scopus_author_id"=>"56374449800"}, {"first_name"=>"Arnold C.", "last_name"=>"Koekman", "scopus_author_id"=>"56373647800"}, {"first_name"=>"Cees", "last_name"=>"Weeterings", "scopus_author_id"=>"23480670100"}, {"first_name"=>"Mark", "last_name"=>"Roest", "scopus_author_id"=>"7004200335"}, {"first_name"=>"Philip G.", "last_name"=>"De Groot", "scopus_author_id"=>"35379095100"}, {"first_name"=>"Eszter", "last_name"=>"Herczenik", "scopus_author_id"=>"16645424200"}, {"first_name"=>"Coen", "last_name"=>"Maas", "scopus_author_id"=>"16199905200"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"doi"=>"10.1371/journal.pone.0108282", "issn"=>"19326203", "sgr"=>"84907495402", "scopus"=>"2-s2.0-84907495402", "pui"=>"600074464", "pmid"=>"25264625", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"8dce5dee-0fb1-3a12-9944-c1bdfa3e5426", "abstract"=>"BACKGROUND:The endocannabinoid system has previously been implicated in the regulation of neurons and inflammatory cells. Additionally, it has been reported that endocannabinoid receptors are present on circulating platelets, but there has been conflicting evidence on their contribution to platelet function.\\n\\nOBJECTIVES:Our aim was to examine the role of endocannabinoids in platelet function in vitro and in vivo.\\n\\nMETHODS AND RESULTS:We studied the effects of the well-characterized endogenous endocannabinoid anandamide on platelet aggregation in suspension, α-granule release, calcium mobilization, Syk phosphorylation, as well as platelet spreading and aggregate formation under flow. Anandamide inhibits platelet aggregation and α-granule release by collagen, collagen-derived peptide CRP-XL, ADP, arachidonic acid and thromboxane A2 analogue U46619. However, activation via thrombin receptor PAR-1 stays largely unaffected. Calcium mobilization is significantly impaired when platelets are stimulated with collagen or CRP-XL, but remains normal in the presence of the other agonists. In line with this finding, we found that anandamide prevents collagen-induced Syk phosphorylation. Furthermore, anandamide-treated platelets exhibit reduced spreading on immobilized fibrinogen, have a decreased capacity for binding fibrinogen in solution and show perturbed platelet aggregate formation under flow over collagen. Finally, we investigated the influence of Cannabis sativa consumption by human volunteers on platelet activation. Similar to our in vitro findings with anandamide, ex vivo collagen-induced platelet aggregation and aggregate formation on immobilized collagen under flow were impaired in whole blood of donors that had consumed Cannabis sativa.\\n\\nCONCLUSIONS:Endocannabinoid receptor agonists reduce platelet activation and aggregate formation both in vitro and ex vivo after Cannabis sativa consumption. Further elucidation of this novel regulatory mechanism for platelet function may prove beneficial in the search for new antithrombotic therapies.", "link"=>"http://www.mendeley.com/research/endocannabinoids-control-platelet-activation-limit-aggregate-formation-under-flow", "reader_count"=>16, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>5, "Student > Postgraduate"=>1, "Other"=>4, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>5, "Student > Postgraduate"=>1, "Other"=>4, "Student > Bachelor"=>3, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>2, "Medicine and Dentistry"=>7, "Neuroscience"=>1, "Social Sciences"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>7}, "Neuroscience"=>{"Neuroscience"=>1}, "Social Sciences"=>{"Social Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "United States"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1695810"], "description"=>"<p>Washed platelets were exposed to a concentration series of anandamide for 40 minutes at 37°C. Subsequently, they were stimulated with 0.5 or 5 µg/mL collagen (A; representative aggregation curves of 0.5 µg/mL collagen stimulation are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108282#pone.0108282.s001\" target=\"_blank\">Figure S1A</a>); 0.1 or 1 µg/mL CRP-XL (B; representative aggregation curves of 0.1 µg/mL CRP-XL are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108282#pone.0108282.s001\" target=\"_blank\">Figure S1B</a>); 10 µM Arachidonic Acid, (AA) (C; representative aggregation curves are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108282#pone.0108282.s001\" target=\"_blank\">Figure S1C</a>); 10 or 100 µM ADP (D; representative curves of 10 µM are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108282#pone.0108282.s001\" target=\"_blank\">Figure S1D</a>); 1 or 10 µM U46619 (E; representative aggregation curves of 1 µM U46619 are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108282#pone.0108282.s001\" target=\"_blank\">Figure S1E</a>); 1, 2.5, 5 or 10 µM TRAP (F; representative aggregation curves of 2.5 µM are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108282#pone.0108282.s001\" target=\"_blank\">Figure S1F</a>), 0.1 U/mL thrombin or anandamide only (G; representative aggregation curves are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108282#pone.0108282.s001\" target=\"_blank\">Figure S1G</a>). Data were normalized to 100% platelet aggregation obtained with the agonists alone in the presence of vehicle. The time-dependence of anandamide inhibition was investigated by incubating washed platelets with 50 µM anandamide or vehicle (DMSO) at 37°C in a time series and subsequently stimulated with 0.5 µg/mL collagen (H). Data are mean ± SD and show 3 or more individual experiments.</p>", "links"=>[], "tags"=>["Platelet activation", "platelet function.ObjectivesOur aim", "Flow BackgroundThe endocannabinoid system", "Endocannabinoids Control Platelet Activation", "Platelet aggregation", "platelet function", "thrombin receptor PAR", "Cannabis sativa.ConclusionsEndocannabinoid receptor agonists", "adp", "2 analogue U 46619.", "Cannabis sativa consumption"], "article_id"=>1186509, "categories"=>["Biological Sciences"], "users"=>["Valentina De Angelis", "Arnold C. Koekman", "Cees Weeterings", "Mark Roest", "Philip G. de Groot", "Eszter Herczenik", "Coen Maas"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0108282.g001", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_endocannabinoid_anandamide_inhibits_platelet_aggregation_/1186509", "title"=>"The endocannabinoid anandamide inhibits platelet aggregation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-29 03:17:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1696019", "https://ndownloader.figshare.com/files/1696022", "https://ndownloader.figshare.com/files/1696024"], "description"=>"<div><p>Background</p><p>The endocannabinoid system has previously been implicated in the regulation of neurons and inflammatory cells. Additionally, it has been reported that endocannabinoid receptors are present on circulating platelets, but there has been conflicting evidence on their contribution to platelet function.</p><p>Objectives</p><p>Our aim was to examine the role of endocannabinoids in platelet function <i>in vitro</i> and <i>in vivo</i>.</p><p>Methods and Results</p><p>We studied the effects of the well-characterized endogenous endocannabinoid anandamide on platelet aggregation in suspension, α-granule release, calcium mobilization, Syk phosphorylation, as well as platelet spreading and aggregate formation under flow. Anandamide inhibits platelet aggregation and α-granule release by collagen, collagen-derived peptide CRP-XL, ADP, arachidonic acid and thromboxane A2 analogue U46619. However, activation via thrombin receptor PAR-1 stays largely unaffected. Calcium mobilization is significantly impaired when platelets are stimulated with collagen or CRP-XL, but remains normal in the presence of the other agonists. In line with this finding, we found that anandamide prevents collagen-induced Syk phosphorylation. Furthermore, anandamide-treated platelets exhibit reduced spreading on immobilized fibrinogen, have a decreased capacity for binding fibrinogen in solution and show perturbed platelet aggregate formation under flow over collagen. Finally, we investigated the influence of <i>Cannabis sativa</i> consumption by human volunteers on platelet activation. Similar to our <i>in vitro</i> findings with anandamide, <i>ex vivo</i> collagen-induced platelet aggregation and aggregate formation on immobilized collagen under flow were impaired in whole blood of donors that had consumed <i>Cannabis sativa</i>.</p><p>Conclusions</p><p>Endocannabinoid receptor agonists reduce platelet activation and aggregate formation both <i>in vitro</i> and <i>ex vivo</i> after <i>Cannabis sativa</i> consumption. Further elucidation of this novel regulatory mechanism for platelet function may prove beneficial in the search for new antithrombotic therapies.</p></div>", "links"=>[], "tags"=>["Platelet activation", "platelet function.ObjectivesOur aim", "Flow BackgroundThe endocannabinoid system", "Endocannabinoids Control Platelet Activation", "Platelet aggregation", "platelet function", "thrombin receptor PAR", "Cannabis sativa.ConclusionsEndocannabinoid receptor agonists", "adp", "2 analogue U 46619.", "Cannabis sativa consumption"], "article_id"=>1186668, "categories"=>["Biological Sciences"], "users"=>["Valentina De Angelis", "Arnold C. Koekman", "Cees Weeterings", "Mark Roest", "Philip G. de Groot", "Eszter Herczenik", "Coen Maas"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0108282.s001", "https://dx.doi.org/10.1371/journal.pone.0108282.s002", "https://dx.doi.org/10.1371/journal.pone.0108282.s003"], "stats"=>{"downloads"=>7, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Endocannabinoids_Control_Platelet_Activation_and_Limit_Aggregate_Formation_under_Flow_/1186668", "title"=>"Endocannabinoids Control Platelet Activation and Limit Aggregate Formation under Flow", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-09-29 03:17:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1695904"], "description"=>"<p>Washed platelets were preincubated with 50 µM anandamide or vehicle for 40 minutes at 37°C. Subsequently, they were exposed to 1 µg/mL collagen, 500 ng/mL CRP-XL, 20 µM ADP, 10 µM U46619, 10 µM TRAP or 1 U/mL thrombin and calcium mobilization was monitored. The (residual) calcium mobilization in the presence of anandamide is expressed as a percentage of uninhibited vehicle control in the presence of the same platelet agonist (A). Western blot analysis of collagen-induced Syk phosphorylation in the presence of vehicle, 50 µM anandamide or control Syk phosphorylation inhibitor PP2 (B; upper lanes indicate phosphorylated Syk (Syk-P), the lower lanes indicate total Syk antigen as a loading control (Syk-T)). Densitometric quantification of collagen-induced Syk-phosphorylation in the presence or absence of anandamide or PP2 (C; n = 4: data are expressed as mean +/− SD).</p>", "links"=>[], "tags"=>["Platelet activation", "platelet function.ObjectivesOur aim", "Flow BackgroundThe endocannabinoid system", "Endocannabinoids Control Platelet Activation", "Platelet aggregation", "platelet function", "thrombin receptor PAR", "Cannabis sativa.ConclusionsEndocannabinoid receptor agonists", "adp", "2 analogue U 46619.", "Cannabis sativa consumption"], "article_id"=>1186573, "categories"=>["Biological Sciences"], "users"=>["Valentina De Angelis", "Arnold C. Koekman", "Cees Weeterings", "Mark Roest", "Philip G. de Groot", "Eszter Herczenik", "Coen Maas"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0108282.g004", "stats"=>{"downloads"=>6, "page_views"=>24, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Anandamide_inhibits_glycoprotein_VI_dependent_calcium_mobilization_and_Syk_phosphorylation_/1186573", "title"=>"Anandamide inhibits glycoprotein VI-dependent calcium mobilization and Syk-phosphorylation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-29 03:17:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1695867"], "description"=>"<p>Platelet α-granule release was studied by flow cytometric analysis of platelet P-selectin externalization. Washed platelets from healthy human donors were pre-exposed to a concentration series of anandamide for 40 min at 37°C and subsequently stimulated with 2.5 µg/mL CRP-XL (A), 2.5 µM U46619 (B) or 5 µM TRAP (C). Data were normalized to the agonist-induced P-selectin expression of vehicle-pretreated control platelets. Reconstituted blood, which was preincubated with anandamide, was perfused over a collagen-coated surface at a shear rate of 1600 s<sup>−1</sup> and snapshots were taken after 5 minutes (D). Subsequently, cover slips were rinsed with buffer for 1 minute to investigate aggregate stability (E). The left panel shows vehicle control, the right panel indicates a perfusion in the presence of 50 µM anandamide. Data are shown as mean ± SD and represent 3 or more individual experiments.</p>", "links"=>[], "tags"=>["Platelet activation", "platelet function.ObjectivesOur aim", "Flow BackgroundThe endocannabinoid system", "Endocannabinoids Control Platelet Activation", "Platelet aggregation", "platelet function", "thrombin receptor PAR", "Cannabis sativa.ConclusionsEndocannabinoid receptor agonists", "adp", "2 analogue U 46619.", "Cannabis sativa consumption"], "article_id"=>1186548, "categories"=>["Biological Sciences"], "users"=>["Valentina De Angelis", "Arnold C. Koekman", "Cees Weeterings", "Mark Roest", "Philip G. de Groot", "Eszter Herczenik", "Coen Maas"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0108282.g002", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Anandamide_inhibits_platelet_945_granule_secretion_and_limits_platelet_aggregate_formation_/1186548", "title"=>"Anandamide inhibits platelet α-granule secretion and limits platelet aggregate formation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-29 03:17:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1695906"], "description"=>"<p>Washed platelets were pretreated with 25 µM FAAH-inhibitor URB597. Subsequently, platelet aggregation was induced by various agonists after a suboptimal exposure to anandamide (10 µM, preincubated for 40 minutes). 0.5 µg/mL collagen (A), 0.1 µg/mL CRP-XL (B), 10 µM AA (C), 10 µM ADP (D), 1 µM U46619 (E) or 1 µM TRAP (F). In further experiments, washed platelets were pretreated with 10 µM indomethacin, exposed to anandamide (10 µM, preincubated for 40 minutes) and stimulated with collagen (0.5 µg/mL: G).</p>", "links"=>[], "tags"=>["Platelet activation", "platelet function.ObjectivesOur aim", "Flow BackgroundThe endocannabinoid system", "Endocannabinoids Control Platelet Activation", "Platelet aggregation", "platelet function", "thrombin receptor PAR", "Cannabis sativa.ConclusionsEndocannabinoid receptor agonists", "adp", "2 analogue U 46619.", "Cannabis sativa consumption"], "article_id"=>1186575, "categories"=>["Biological Sciences"], "users"=>["Valentina De Angelis", "Arnold C. Koekman", "Cees Weeterings", "Mark Roest", "Philip G. de Groot", "Eszter Herczenik", "Coen Maas"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0108282.g005", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_inhibitory_effect_of_anandamide_on_platelet_activation_is_not_dependent_on_platelet_preactivation_/1186575", "title"=>"The inhibitory effect of anandamide on platelet activation is not dependent on platelet preactivation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-29 03:17:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1695916"], "description"=>"<p>Whole blood from <i>Cannabis sativa</i> consumers (n = 4; “Consumers”) or healthy control donors (“Controls”) was perfused for 5 minutes over immobilized collagen at a shear rate of 1600 s<sup>−1</sup> (A). In further experiments, collagen-induced platelet aggregation was investigated in platelet-rich plasma from these <i>Cannabis sativa</i> consumers and controls (B). Representative aggregation curves are shown in panel C. Finally, collagen-induced platelet aggregation was studied in platelet-rich plasma of three self-reported <i>Cannabis sativa</i> consumers on two separate instances: once after 10 days of daily consumption of <i>Cannabis sativa</i> (“Use”), as well as after a period of 10 days without consumption (“Withdrawal”).</p>", "links"=>[], "tags"=>["Platelet activation", "platelet function.ObjectivesOur aim", "Flow BackgroundThe endocannabinoid system", "Endocannabinoids Control Platelet Activation", "Platelet aggregation", "platelet function", "thrombin receptor PAR", "Cannabis sativa.ConclusionsEndocannabinoid receptor agonists", "adp", "2 analogue U 46619.", "Cannabis sativa consumption"], "article_id"=>1186585, "categories"=>["Biological Sciences"], "users"=>["Valentina De Angelis", "Arnold C. Koekman", "Cees Weeterings", "Mark Roest", "Philip G. de Groot", "Eszter Herczenik", "Coen Maas"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0108282.g006", "stats"=>{"downloads"=>2, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cannabis_sativa_consumption_limits_platelet_aggregate_formation_under_flow_and_reduces_platelet_responsiveness_to_collagen_/1186585", "title"=>"<i>Cannabis sativa</i> consumption limits platelet aggregate formation under flow and reduces platelet responsiveness to collagen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-29 03:17:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1695880"], "description"=>"<p>Washed platelets from healthy human donors were exposed to a concentration series (0–50 µM) of anandamide for 40 minutes at 37°C and subsequently stimulated with 2.5 µg/mL CRP-XL (A), 2.5 µM U46619 (B) or 5 µM TRAP (C). Platelet GPIIb/IIIa activation was studied by flow cytometry. Data were normalized to the agonist-induced GPIIb/IIIa activation of vehicle-pretreated control platelets. Washed platelets were preincubated with 50 µM anandamide or vehicle for 40 minutes at 37°C and perfused over immobilized fibrinogen- at a shear rate of 25 s<sup>−1</sup>. Subsequently, differential interference contrast microscopy images were taken (D; representative images after 15 minutes of perfusion, scale bars represent 10 µm). Quantification of the spreading behaviour of individual platelets on immobilized fibrinogen (E; spreading was quantified for 3 individual platelets per separate experiment). Quantification of the amount of adhering platelets per field (F). Data represent mean and standard deviation (SD) of 3 or more individual experiments.</p>", "links"=>[], "tags"=>["Platelet activation", "platelet function.ObjectivesOur aim", "Flow BackgroundThe endocannabinoid system", "Endocannabinoids Control Platelet Activation", "Platelet aggregation", "platelet function", "thrombin receptor PAR", "Cannabis sativa.ConclusionsEndocannabinoid receptor agonists", "adp", "2 analogue U 46619.", "Cannabis sativa consumption"], "article_id"=>1186561, "categories"=>["Biological Sciences"], "users"=>["Valentina De Angelis", "Arnold C. Koekman", "Cees Weeterings", "Mark Roest", "Philip G. de Groot", "Eszter Herczenik", "Coen Maas"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0108282.g003", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Anandamide_reduces_glycoprotein_IIb_IIIa_activation_and_inhibits_platelet_spreading_/1186561", "title"=>"Anandamide reduces glycoprotein IIb/IIIa activation and inhibits platelet spreading.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-29 03:17:12"}

PMC Usage Stats | Further Information

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Relative Metric

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