{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T14:13:50Z","timestamp":1788444830160,"version":"build-2803163510"},"reference-count":43,"publisher":"IMR Press","issue":"8","license":[{"start":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:00:00Z","timestamp":1786579200000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmazie"],"abstract":"<jats:p>Background:Aberrant platelet activation is a major contributor to thrombotic disorders, prompting interest in naturally derived candidates that can safely regulate platelet function. Sappanchalcone, a chalcone-type flavonoid derived from Caesalpinia sappan L., has diverse biological activities; however, its effects on platelet activation and clot retraction remain unclear. This study evaluated the antiplatelet potential of sappanchalcone and its underlying mechanisms in human platelets.Methods:Washed human platelets and platelet-rich plasma were used to assess platelet activation and clot retraction, respectively. Samples were pretreated with sappanchalcone (16\u2013128 \u03bcM, depending on the assay) and stimulated with collagen or thrombin. Platelet aggregation and cytotoxicity were assessed by light transmission aggregometry and lactate dehydrogenase (LDH) leakage assay. cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), and thromboxane B2 (TXB2) levels were quantified using enzyme-linked immunosorbent assay (ELISA) kits, and intracellular Ca2+ mobilization was measured by Fura-2\/AM-based fluorescence analysis. Protein phosphorylation was examined by Western blotting, whereas integrin \u03b1IIb\u03b23\u2013fibrinogen binding was analyzed by flow cytometry. Dense granule secretion was evaluated using adenosine triphosphate (ATP) luminescence and serotonin ELISA assays. Thrombin-induced clot retraction was quantified by ImageJ (v1.54k, U.S. National Institutes of Health, Bethesda, MD, USA)-based residual clot area analysis. Data are presented as mean \u00b1 standard deviation (SD) and analyzed by one-way analysis of variance (ANOVA) with Tukey\u2013Kramer post hoc test (n = 4; p &lt; 0.05).Results:Sappanchalcone concentration-dependently inhibited collagen-induced platelet aggregation, reducing aggregation from 83.0% to 10.7% at 128 \u03bcM, with a half-maximal inhibitory concentration (IC50) of 57.16 \u03bcM, without cytotoxicity. At 128 \u03bcM, sappanchalcone increased cAMP from 7.02 \u00b1 0.85 to 11.96 \u00b1 0.57 pmol\/108 platelets and reduced Ca2+ mobilization from 711.97 \u00b1 47.29 to 66.11 \u00b1 6.83 nM. Vasodilator-stimulated phosphoprotein (VASP) Ser157 and Ser239 phosphorylation increased by 2.97- and 2.07-fold, respectively, whereas fibrinogen binding decreased from 74.43 \u00b1 5.67% to 21.90 \u00b1 1.25%. Phosphoinositide 3-kinase (PI3K), protein kinase B (Akt), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 phosphorylation was reduced to 0.69-, 0.21-, 1.81-, 0.27-, and 0.49-fold, respectively. ATP secretion, serotonin release, TXB2 levels reflecting thromboxane A2 (TXA2) generation, cytosolic phospholipase A2 (cPLA2) phosphorylation, and thrombin-induced clot retraction were also suppressed.Conclusions:Sappanchalcone limits platelet activation, aggregation, and clot retraction by enhancing cyclic nucleotide-mediated inhibitory signaling and suppressing PI3K\/Akt and mitogen-activated protein kinase (MAPK) phosphorylation. These findings provide mechanistic evidence supporting sappanchalcone as a potential naturally derived antiplatelet candidate.<\/jats:p>","DOI":"10.31083\/pharmazie54071","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T13:26:12Z","timestamp":1788441972000},"source":"Crossref","is-referenced-by-count":0,"title":["Sappanchalcone Attenuates Platelet Activation and Clot Retraction Through Coordinated Modulation of Cyclic Nucleotides and PI3K\/MAPK Signaling Pathways"],"prefix":"10.31083","volume":"81","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-8841-4176","authenticated-orcid":true,"given":"Ga Hee","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Biomedical Laboratory Science, Namseoul University, 31020 Cheonan, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-0878-6393","authenticated-orcid":true,"given":"Jin Pyo","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Biomedical Laboratory Science, Namseoul University, 31020 Cheonan, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4259-7928","authenticated-orcid":true,"given":"Chang Eun","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Biomedical Laboratory Science, Namseoul University, 31020 Cheonan, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7042-391X","authenticated-orcid":true,"given":"Dong-Ha","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Biomedical Laboratory Science, Namseoul University, 31020 Cheonan, Chungcheongnam-do, Republic of Korea"},{"name":"Molecular Diagnostics Research Institute, Namseoul University, 31020 Cheonan, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,8,13]]},"reference":[{"key":"ref1","unstructured":"World Health Organization. Cardiovascular diseases (CVDs). WHO Fact Sheet. 2021. Available at: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/cardiovascular-diseases-(cvds) (Accessed: 11 June 2021)."},{"key":"ref2","doi-asserted-by":"publisher","unstructured":"Badimon L, Padr\u00f3 T, Vilahur G. Atherosclerosis, platelets and thrombosis in acute ischaemic heart disease. European Heart Journal. Acute Cardiovascular Care. 2012; 1: 60\u201374.","DOI":"10.1177\/2048872612441582"},{"key":"ref3","doi-asserted-by":"publisher","unstructured":"Tsao CW, Aday AW, Almarzooq ZI, Anderson CAM, Arora P, Avery CL, et al. Heart Disease and Stroke Statistics-2023 Update: A Report From the American Heart Association. Circulation. 2023; 147: e93\u2013e621.","DOI":"10.1161\/CIR.0000000000001123"},{"key":"ref4","doi-asserted-by":"publisher","unstructured":"Randeni N, Luo J, Xu B. New insights into anti-thrombotic effects of dietary bioactive components. Food & Function. 2025; 16: 8695\u20138719.","DOI":"10.1039\/d5fo01827g"},{"key":"ref5","doi-asserted-by":"publisher","unstructured":"Stark K, Massberg S. Interplay between inflammation and thrombosis in cardiovascular pathology. Nature Reviews. Cardiology. 2021; 18: 666\u2013682.","DOI":"10.1038\/s41569-021-00552-1"},{"key":"ref6","doi-asserted-by":"publisher","unstructured":"Varga-Szabo D, Braun A, Nieswandt B. Calcium signaling in platelets. Journal of Thrombosis and Haemostasis: JTH. 2009; 7: 1057\u20131066.","DOI":"10.1111\/j.1538-7836.2009.03455.x"},{"key":"ref7","doi-asserted-by":"publisher","unstructured":"Li Z, Delaney MK, O'Brien KA, Du X. Signaling during platelet adhesion and activation. Arteriosclerosis, Thrombosis, and Vascular Biology. 2010; 30: 2341\u20132349.","DOI":"10.1161\/ATVBAHA.110.207522"},{"key":"ref8","doi-asserted-by":"publisher","unstructured":"Lin LL, Wartmann M, Lin AY, Knopf JL, Seth A, Davis RJ. cPLA2 is phosphorylated and activated by MAP kinase. Cell. 1993; 72: 269\u2013278.","DOI":"10.1016\/0092-8674(93)90666-e"},{"key":"ref9","doi-asserted-by":"publisher","unstructured":"Akram AW, Lee GH, Baek SM, Kang J, Koo Y, Oh Y, et al. Xanthium strumarium L. Exhibits Potent Antiplatelet and Antithrombotic Effects by Modulating MAPK and PI3K\/AKT Signaling Pathways and Inhibiting Ferric Chloride-Induced Thrombosis. Biomedicines. 2025; 13: 2924.","DOI":"10.3390\/biomedicines13122924"},{"key":"ref10","doi-asserted-by":"publisher","unstructured":"Huang J, Li X, Shi X, Zhu M, Wang J, Huang S, et al. Platelet integrin \u03b1IIb\u03b23: signal transduction, regulation, and its therapeutic targeting. Journal of Hematology & Oncology. 2019; 12: 26.","DOI":"10.1186\/s13045-019-0709-6"},{"key":"ref11","doi-asserted-by":"publisher","unstructured":"Niu H, Chen X, Gruppo RA, Li D, Wang Y, Zhang L, et al. Integrin \u03b1IIb-mediated PI3K\/Akt activation in platelets. PLoS ONE. 2012; 7: e47356.","DOI":"10.1371\/journal.pone.0047356"},{"key":"ref12","doi-asserted-by":"publisher","unstructured":"Jansen EE, Hartmann M. Clot Retraction: Cellular Mechanisms and Inhibitors, Measuring Methods, and Clinical Implications. Biomedicines. 2021; 9: 1064.","DOI":"10.3390\/biomedicines9081064"},{"key":"ref13","doi-asserted-by":"publisher","unstructured":"Bye AP, Unsworth AJ, Gibbins JM. Platelet signaling: a complex interplay between inhibitory and activatory networks. Journal of Thrombosis and Haemostasis: JTH. 2016; 14: 918\u2013930.","DOI":"10.1111\/jth.13302"},{"key":"ref14","doi-asserted-by":"publisher","unstructured":"Shpakova V, Rukoyatkina N, Walter U, Gambaryan S. Potential and limitations of PKA\/ PKG inhibitors for platelet studies. Platelets. 2022; 33: 859\u2013868.","DOI":"10.1080\/09537104.2021.2003316"},{"key":"ref15","doi-asserted-by":"publisher","unstructured":"Rajput MS, Nirmal NP, Nirmal SJ, Santivarangkna C. Bio-actives from Caesalpinia sappan L.: recent advancements in phytochemistry and pharmacology. South African Journal of Botany. 2022; 151: 60\u201374.","DOI":"10.1016\/j.sajb.2021.11.021"},{"key":"ref16","doi-asserted-by":"publisher","unstructured":"Asevedo EA, Ramos Santiago L, Kim HJ, Syahputra RA, Park MN, Ribeiro RIMA, et al. Unlocking the therapeutic mechanism of Caesalpinia sappan: a comprehensive review of its antioxidant and anti-cancer properties, ethnopharmacology, and phytochemistry. Frontiers in Pharmacology. 2025; 15: 1514573.","DOI":"10.3389\/fphar.2024.1514573"},{"key":"ref17","doi-asserted-by":"publisher","unstructured":"Jung EG, Han KI, Kwon HJ, Patnaik BB, Kim WJ, Hur GM, et al. Anti-inflammatory activity of sappanchalcone isolated from Caesalpinia sappan L. in a collagen-induced arthritis mouse model. Archives of Pharmacal Research. 2015; 38: 973\u2013983.","DOI":"10.1007\/s12272-015-0557-z"},{"key":"ref18","doi-asserted-by":"publisher","unstructured":"Seo HW, No H, Cheon HJ, Kim JK. Sappanchalcone, a flavonoid isolated from Caesalpinia sappan L., induces caspase-dependent and AIF-dependent apoptosis in human colon cancer cells. Chemico-biological Interactions. 2020; 327: 109185.","DOI":"10.1016\/j.cbi.2020.109185"},{"key":"ref19","doi-asserted-by":"publisher","unstructured":"Jantan I, Raweh SM, Sirat HM, Jamil S, Mohd Yasin YH, Jalil J, et al. Inhibitory effect of compounds from Zingiberaceae species on human platelet aggregation. Phytomedicine : International Journal of Phytotherapy and Phytopharmacology. 2008; 15: 306\u2013309.","DOI":"10.1016\/j.phymed.2007.08.002"},{"key":"ref20","doi-asserted-by":"publisher","unstructured":"Lien LM, Lin KH, Huang LT, Tseng MF, Chiu HC, Chen RJ, et al. Licochalcone A Prevents Platelet Activation and Thrombus Formation through the Inhibition of PLC\u03b32-PKC, Akt, and MAPK Pathways. International Journal of Molecular Sciences. 2017; 18: 1500.","DOI":"10.3390\/ijms18071500"},{"key":"ref21","doi-asserted-by":"publisher","unstructured":"Nam GS. Antiplatelet Effects of Taraxacum mongolicum Ethanol Extract through Modulation of Collagen-dependent Platelet Activation. Biomedical Science Letters. 2026; 32: 88\u201397.","DOI":"10.15616\/BSL.2026.32.1.88"},{"key":"ref22","doi-asserted-by":"publisher","unstructured":"Lee DH. Anti-platelet Effects of Artemisinin through Regulation of Cyclic Nucleotide on Collagen-induced human Platelets. Biomedical Science Letters. 2024; 30: 162\u2013168.","DOI":"10.15616\/BSL.2024.30.3.162"},{"key":"ref23","doi-asserted-by":"publisher","unstructured":"Lee DH. Antiplatelet Effect of Chrysoeriol by Regulating cAMP and cGMP in U46619-induced Human Platelets. Biomedical Science Letters. 2025; 31: 254\u2013261.","DOI":"10.15616\/BSL.2025.31.3.254"},{"key":"ref24","doi-asserted-by":"publisher","unstructured":"Lee GH, Lee JP, Wahab AA, Heo NY, Park CE, Lee DH. Chrysoeriol Exerts Antiplatelet Effects by Regulating cAMP\/cGMP and PI3K\/MAPK Pathway. Biomolecules & Therapeutics. 2026; 34: 202\u2013212.","DOI":"10.4062\/biomolther.2025.090"},{"key":"ref25","doi-asserted-by":"publisher","unstructured":"Marjoram RJ, Li Z, He L, Tollefsen DM, Kunicki TJ, Dickeson SK, et al. \u03b12\u03b21 integrin, GPVI receptor, and common FcR\u03b3 chain on mouse platelets mediate distinct responses to collagen in models of thrombosis. PLoS ONE. 2014; 9: e114035.","DOI":"10.1371\/journal.pone.0114035"},{"key":"ref26","doi-asserted-by":"publisher","unstructured":"Smolenski A. Novel roles of cAMP\/cGMP-dependent signaling in platelets. Journal of Thrombosis and Haemostasis: JTH. 2012; 10: 167\u2013176.","DOI":"10.1111\/j.1538-7836.2011.04576.x"},{"key":"ref27","doi-asserted-by":"publisher","unstructured":"Hochuli R, Dicenta V, Laspa Z, Sigle M, Harm T, Castor T, et al. Effect of phosphodiesterase inhibitors on platelet function. Biochemistry and Biophysics Reports. 2025; 43: 102115.","DOI":"10.1016\/j.bbrep.2025.102115"},{"key":"ref28","doi-asserted-by":"crossref","unstructured":"Butt E, Abel K, Krieger M, Palm D, Hoppe V, Hoppe J, et al. cAMP- and cGMP-dependent protein kinase phosphorylation sites of the focal adhesion vasodilator-stimulated phosphoprotein (VASP) in vitro and in intact human platelets. The Journal of Biological Chemistry. 1994; 269: 14509\u201314517.","DOI":"10.1016\/S0021-9258(17)36652-8"},{"key":"ref29","doi-asserted-by":"publisher","unstructured":"Joo SJ. Mechanisms of Platelet Activation and Integrin \u03b1II\u03b23. Korean Circulation Journal. 2012; 42: 295\u2013301.","DOI":"10.4070\/kcj.2012.42.5.295"},{"key":"ref30","doi-asserted-by":"publisher","unstructured":"Litvinov RI, Farrell DH, Weisel JW, Bennett JS. The Platelet Integrin \u03b1IIb\u03b23 Differentially Interacts with Fibrin Versus Fibrinogen. The Journal of Biological Chemistry. 2016; 291: 7858\u20137867.","DOI":"10.1074\/jbc.M115.706861"},{"key":"ref31","doi-asserted-by":"crossref","unstructured":"Seger R, Krebs EG. The MAPK signaling cascade. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology. 1995; 9: 726\u2013735.","DOI":"10.1096\/fasebj.9.9.7601337"},{"key":"ref32","doi-asserted-by":"publisher","unstructured":"Minuz P, Gaino S, Zuliani V, Tommasoli RM, Benati D, Ortolani R, et al. Functional role of p38 mitogen activated protein kinase in platelet activation induced by a thromboxane A2 analogue and by 8-iso-prostaglandin F2alpha. Thrombosis and Haemostasis. 2002; 87: 888\u2013898.","DOI":"10.1055\/s-0037-1613101"},{"key":"ref33","doi-asserted-by":"publisher","unstructured":"Adam F, Kauskot A, Nurden P, Sulpice E, Hoylaerts MF, Davis RJ, et al. Platelet JNK1 is involved in secretion and thrombus formation. Blood. 2010; 115: 4083\u20134092.","DOI":"10.1182\/blood-2009-07-233932"},{"key":"ref34","doi-asserted-by":"publisher","unstructured":"Holinstat M, Boutaud O, Apopa PL, Vesci J, Bala M, Oates JA, et al. Protease-activated receptor signaling in platelets activates cytosolic phospholipase A2\u03b1 differently for cyclooxygenase-1 and 12-lipoxygenase catalysis. Arteriosclerosis, Thrombosis, and Vascular Biology. 2011; 31: 435\u2013442.","DOI":"10.1161\/ATVBAHA.110.219527"},{"key":"ref35","doi-asserted-by":"publisher","unstructured":"Li Z, Zhang G, Le Breton GC, Gao X, Malik AB, Du X. Two waves of platelet secretion induced by thromboxane A2 receptor and a critical role for phosphoinositide 3-kinases. The Journal of Biological Chemistry. 2003; 278: 30725\u201330731.","DOI":"10.1074\/jbc.M301838200"},{"key":"ref36","doi-asserted-by":"publisher","unstructured":"Akram AW, Choi DC, Chae HK, Kim SD, Kwak D, Yun BS, et al. Dihydrogeodin from Fennellia flavipes Modulates Platelet Aggregation via Downregulation of Calcium Signaling, \u03b1IIb\u03b23 Integrins, MAPK, and PI3K\/Akt Pathways. Marine Drugs. 2025; 23: 212.","DOI":"10.3390\/md23050212"},{"key":"ref37","doi-asserted-by":"publisher","unstructured":"Shin JH, Akram AW, Saba E, Kim MC, Kim SD, Park SJ, et al. Ginsenoside Rs3 (G-Rs3) inhibits agonist- and oxLDL-mediated platelet activation and attenuates in vivo arterial thrombosis. Journal of Ginseng Research. 2026; 50: 101020.","DOI":"10.1016\/j.jgr.2026.101020"},{"key":"ref38","doi-asserted-by":"publisher","unstructured":"Guo F, Yang R, Xiong X, Yuan Y, Zhang L, Hua C, et al. Taxifolin Inhibits Platelet Activation and Thrombosis by Regulating the PI3K\/Akt and MAPK Signaling Pathways. Molecular Nutrition & Food Research. 2025; 69: e70129.","DOI":"10.1002\/mnfr.70129"},{"key":"ref39","doi-asserted-by":"publisher","unstructured":"Balykina A, Naida L, Kirkg\u00f6z K, Nikolaev VO, Fock E, Belyakov M, et al. Antiplatelet Effects of Flavonoid Aglycones Are Mediated by Activation of Cyclic Nucleotide-Dependent Protein Kinases. International Journal of Molecular Sciences. 2024; 25: 4864.","DOI":"10.3390\/ijms25094864"},{"key":"ref40","doi-asserted-by":"publisher","unstructured":"Tutwiler V, Litvinov RI, Lozhkin AP, Peshkova AD, Lebedeva T, Ataullakhanov FI, et al. Kinetics and mechanics of clot contraction are governed by the molecular and cellular composition of the blood. Blood. 2016; 127: 149\u2013159.","DOI":"10.1182\/blood-2015-05-647560"},{"key":"ref41","doi-asserted-by":"publisher","unstructured":"Weng Q, Chen L, Ye L, Lu X, Yu Z, Wen C, et al. Determination of licochalcone A in rat plasma by UPLC\u2013MS\/MS and its pharmacokinetics. Acta Chromatographica. 2019; 31: 262\u2013265.","DOI":"10.1556\/1326.2018.00491"},{"key":"ref42","doi-asserted-by":"publisher","unstructured":"Rudrapal M, Khan J, Dukhyil AAB, Alarousy RMII, Attah EI, Sharma T, et al. Chalcone Scaffolds, Bioprecursors of Flavonoids: Chemistry, Bioactivities, and Pharmacokinetics. Molecules (Basel, Switzerland). 2021; 26: 7177.","DOI":"10.3390\/molecules26237177"},{"key":"ref43","doi-asserted-by":"publisher","unstructured":"Salunkhe R, Gadgoli C, Naik A, Patil N. Pharmacokinetic Profile and Oral Bioavailability of Diosgenin, Charantin, and Hydroxychalcone From a Polyherbal Formulation. 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