Albiflorin (AF), a primary bioactive compound from the roots of Paeonia lactiflora Pall. (Peony), demonstrates anti-inflammatory, antioxidant, and cytoprotective effects. This study aims to investigate the antithrombotic potential of AF and clarify its mechanisms of action. Its anticoagulant effects were assessed via activated partial thromboplastin time, prothrombin time, thrombin, and factor Xa (FXa) activities, as well as fibrin formation and platelet aggregation. Additionally, the effect of plasminogen activator inhibitor type 1 and tissue-type plasminogen activator was evaluated in tumor necrosis factor-α-stimulated human umbilical vein endothelial cells (HUVECs). Our experiments revealed that AF exhibited antithrombotic activity comparable to that of rivaroxaban, a well-established direct FXa inhibitor, particularly in suppressing FXa activity and platelet aggregation induced by adenosine diphosphate (ADP) and U46619, a synthetic thromboxane A2 analog. AF also reduced the surface expression of P-selectin, inhibited phosphorylation of myristoylated alanine-rich C kinase substrate, and decreased PAC-1 activation following ADP or U46619 stimulation. Furthermore, AF enhanced nitric oxide production while preventing excess endothelin-1 release in HUVECs exposed to these agonists. In vivo experiments in mouse models of arterial and pulmonary thrombosis demonstrated that AF is a potent anticoagulant and antithrombotic agent. These findings indicate that AF could serve as a promising lead compound for developing innovative anti-FXa and antiplatelet therapeutics.
GongG, QinY, HuangW. Anti-thrombosis effect of diosgenin extract from Dioscorea zingiberensis C.H. Wright in vitro and in vivo. Phytomedicine, 2011; 18(6):458–463; doi: 10.1016/j.phymed.2010.08.015
2.
RobertiR, IannoneLF, PalleriaC, et al.Direct oral anticoagulants: From randomized clinical trials to real-world clinical practice. Front Pharmacol, 2021; 12:684638; doi: 10.3389/fphar.2021.684638
3.
MillingTJJr, ZiebellCM. A review of oral anticoagulants, old and new, in major bleeding and the need for urgent surgery. Trends Cardiovasc Med, 2020; 30(2):86–90; doi: 10.1016/j.tcm.2019.03.004
4.
AbdelnabiM, BenjanuwattraJ, OkashaO, et al.Switching from warfarin to direct-acting oral anticoagulants: It is time to move forward!. Egypt Heart J, 2022; 74(1):18; doi: 10.1186/s43044-022-00259-9
5.
RoziW, RahhalA, AliEA, et al.Direct oral anticoagulants in sickle cell disease: A systematic review and meta-analysis. Blood Adv, 2022; 6(17):5061–5066; doi: 10.1182/bloodadvances.2022007308
6.
Rustem GulluogluF, SouvereinPC, van den HamHA, et al.Comparative effectiveness and safety of direct oral anticoagulants versus warfarin in UK patients with atrial fibrillation and type 2 diabetes: A retrospective cohort study. Pharmacoepidemiol Drug Saf, 2021; 30(10):1293–1320; doi: 10.1002/pds.5181
7.
DobeshPP, KernanMM, LueshenJJ. Direct oral anticoagulants in the treatment of venous thromboembolism: Use in patients with advanced renal impairment, obesity, or other weight-related special populations. Semin Respir Crit Care Med, 2021; 42(2):233–249; doi: 10.1055/s-0041-1723952
8.
BavaliaR, MiddeldorpS, WeisserG, et al.Treatment of venous thromboembolism in special populations with direct oral anticoagulants. Thromb Haemost, 2020; 120(6):899–911; doi: 10.1055/s-0040-1710314
9.
ZelnikerTA, RuffCT, AntmanEM, et al.The efficacy and safety of non-vitamin K antagonist oral anticoagulants in patients with atrial fibrillation and coronary artery disease: A meta-analysis of randomized trials. Eur Heart J Acute Cardiovasc Care, 2019; 8(6):554–561; doi: 10.1177/2048872618796990
10.
GrymonprezM, SimoensC, SteurbautS, et al.Worldwide trends in oral anticoagulant use in patients with atrial fibrillation from 2010 to 2018: A systematic review and meta-analysis. Europace, 2022; 24(6):887–898; doi: 10.1093/europace/euab303
11.
YiYH, GongS, GongTL, et al.New oral anticoagulants for venous thromboembolism prophylaxis in total hip and knee arthroplasty: A systematic review and network meta-analysis. Front Pharmacol, 2021; 12:775126; doi: 10.3389/fphar.2021.775126
12.
KupoP, SzakacsZ, SolymarM, et al.Direct anticoagulants and risk of myocardial infarction, a multiple treatment network meta-analysis. Angiology, 2020; 71(1):27–37; doi: 10.1177/0003319719874255
13.
LopesRD, HeizerG, AronsonR, et al.; AUGUSTUS Investigators. Antithrombotic therapy after acute coronary syndrome or pci in atrial fibrillation. N Engl J Med, 2019; 380(16):1509–1524; doi: 10.1056/NEJMoa1817083
14.
SzaparyL, TornyosD, KupoP, et al.Combination of antiplatelet and anticoagulant therapy, component network meta-analysis of randomized controlled trials. Front Cardiovasc Med, 2022; 9:1036609; doi: 10.3389/fcvm.2022.1036609
15.
RuggeriZM. Platelets in atherothrombosis. Nat Med, 2002; 8(11):1227–1234; doi: 10.1038/nm1102-1227
JohansenM. Antiplatelet therapy after aspirin-induced upper gastrointestinal bleeding. Tidsskr Nor Laegeforen, 2006; 126(21):2802–2804; doi: 10.1016/j.ejim.2011.10.004
18.
LiHX, HanSY, WangXW, et al.Effect of the carthamins yellow from Carthamus tinctorius L. on hemorheological disorders of blood stasis in rats. Food Chem Toxicol, 2009; 47(8):1797–1802; doi: 10.1016/j.fct.2009.04.026
19.
LiuL, DuanJA, TangY, et al.Taoren-Honghua herb pair and its main components promoting blood circulation through influencing on hemorheology, plasma coagulation and platelet aggregation. J Ethnopharmacol, 2012; 139(2):381–387; doi: 10.1016/j.jep.2011.11.016
20.
ZhangH, WangJ, LangW, et al.Albiflorin ameliorates inflammation and oxidative stress by regulating the NF-kappaB/NLRP3 pathway in Methotrexate-induced enteritis. Int Immunopharmacol, 2022; 109:108824; doi: 10.1016/j.intimp.2022.108824
21.
LiuP, ChenJ, MaS, et al.Albiflorin attenuates mood disorders under neuropathic pain state by suppressing the hippocampal NLRP3 inflammasome activation during chronic constriction injury. Int J Neuropsychopharmacol, 2021; 24(1):64–76; doi: 10.1093/ijnp/pyaa076
22.
KimC, RyuSH, KimN, et al.Renal protective effects of Sparstolonin B in a mouse model of sepsis. Biotechnol Bioproc E, 2022; 27(2):157–162; doi: 10.1007/s12257-021-0319-3
23.
LeeIC, BaeJS. Hepatic protective effects of Jujuboside B through the modulation of inflammatory pathways. Biotechnol Bioproc E, 2022; 27(3):336–343; doi: 10.1007/s12257-022-0049-1
24.
KimN, JeonC, KimC, et al.Inhibition of factor Xa activity, platelet aggregation, and experimentally induced thrombosis by Sparstolonin B. Phytomedicine, 2022; 99:153987; doi: 10.1016/j.phymed.2022.153987
25.
HahnD, KimMJ, KwonY, et al.Natural products ameliorating the adverse health effects by air particulate matter. Biotechnol Bioproc E, 2024; 29(1):1–24; doi: 10.1007/s12257-024-00012-w
26.
WuCC, WuCI, WangWY, et al.Low concentrations of resveratrol potentiate the antiplatelet effect of prostaglandins. Planta Med, 2007; 73(5):439–443; doi: 10.1055/s-2007-967173
27.
GrynkiewiczG, PoenieM, TsienRY. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem, 1985; 260(6):3440–3450; doi: 10.1016/S0021-9258(19)83641-4; doi: 3838314
28.
FrojmovicM, WongT, van de VenT. Dynamic measurements of the platelet membrane glycoprotein IIb-IIIa receptor for fibrinogen by flow cytometry. I. Methodology, theory and results for two distinct activators. Biophys J, 1991; 59(4):815–827; doi: 10.1016/S0006-3495(91)82294-9
29.
KimGO, HeoJB, ParkDH, et al.Antiplatelet aggregation properties of cirsilineol: A novel inhibitor of blood coagulation factor Xa. Pharmaceuticals (Basel), 2023; 16(4):588; doi: 10.3390/ph16040588
30.
LeeW, LeeD, LeeY, et al.Isolation, synthesis, and antisepsis effects of a C-methylcoumarinochromone isolated from Abronia nana cell culture. J Nat Prod, 2018; 81(5):1173–1182; doi: 10.1021/acs.jnatprod.7b00826
31.
PerzbornE, RoehrigS, StraubA, et al.Rivaroxaban: A new oral factor Xa inhibitor. Arterioscler Thromb Vasc Biol, 2010; 30(3):376–381; doi: 10.1161/ATVBAHA.110.202978
32.
LeeW, KuSK, KimMA, et al.Anti-factor Xa activities of zingerone with anti-platelet aggregation activity. Food Chem Toxicol, 2017; 105:186–193; doi: 10.1016/j.fct.2017.04.012
33.
IzuharaY, TakahashiS, NangakuM, et al.Inhibition of plasminogen activator inhibitor-1: Its mechanism and effectiveness on coagulation and fibrosis. Arterioscler Thromb Vasc Biol, 2008; 28(4):672–677; doi: 10.1161/ATVBAHA.107.157479
34.
HahnD, BaeJS. Recent progress in the discovery of bioactive components from edible natural sources with antithrombotic activity. J Med Food, 2019; 22(2):109–120; doi: 10.1089/jmf.2018.4268
35.
PetzoldT, ThienelM, DannenbergL, et al.Rivaroxaban reduces arterial thrombosis by inhibition of FXa-driven platelet activation via protease activated receptor-1. Circ Res, 2020; 126(4):486–500; doi: 10.1161/CIRCRESAHA.119.315099
36.
ElzagallaaiA, RoseSD, TrifaroJM. Platelet secretion induced by phorbol esters stimulation is mediated through phosphorylation of MARCKS: A MARCKS-derived peptide blocks MARCKS phosphorylation and serotonin release without affecting pleckstrin phosphorylation. Blood, 2000; 95(3):894–902; doi: 10.1182/blood.V95.3.894.003k15_894_902
37.
WeiAH, SchoenwaelderSM, AndrewsRK, et al.New insights into the haemostatic function of platelets. Br J Haematol, 2009; 147(4):415–430; doi: 10.1111/j.1365-2141.2009.07819.x
38.
MertenM, ThiagarajanP. P-selectin expression on platelets determines size and stability of platelet aggregates. Circulation, 2000; 102(16):1931–1936; doi: 10.1161/01.cir.102.16.1931
39.
BredtDS, SnyderSH. Nitric oxide: A physiologic messenger molecule. Annu Rev Biochem, 1994; 63:175–195; doi: 10.1146/annurev.biochem.63.1.175
40.
YanagisawaM, KuriharaH, KimuraS, et al.A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature, 1988; 332(6163):411–415; doi: 10.1038/332411a0
41.
d’UscioLV, BartonM, ShawS, et al.Endothelin in atherosclerosis: Importance of risk factors and therapeutic implications. J Cardiovasc Pharmacol, 2000; 35(4 (Suppl 2)):S55–S59; doi: 10.1097/00005344-200000002-00013
42.
Schmitz-SpankeS, SchipkeJD. Potential role of endothelin-1 and endothelin antagonists in cardiovascular diseases. Basic Res Cardiol, 2000; 95(4):290–298; doi: 10.1007/s003950070048
43.
VerhammeP, HoylaertsMF. The pivotal role of the endothelium in haemostasis and thrombosis. Acta Clin Belg, 2006; 61(5):213–219; doi: 10.1179/acb.2006.036
44.
FerrarisVA, FerrarisSP, SahaSP. Antiplatelet drugs: Mechanisms and risks of bleeding following cardiac operations. Int J Angiol, 2011; 20(1):1–18; doi: 10.1055/s-0031-1272544
45.
EikelboomJW, HirshJ, SpencerFA, et al.Antiplatelet drugs: Antithrombotic therapy and prevention of thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines. Chest, 2012; 141(2 Suppl):e89S–e119S; doi: 10.1378/chest.11-2293
LoffredoL, PerriL, VioliF. Myocardial infarction and atrial fibrillation: Different impact of anti-IIa vs anti-Xa new oral anticoagulants: A meta-analysis of the interventional trials. Int J Cardiol, 2015; 178:8–9; doi: 10.1016/j.ijcard.2014.10.124
48.
BauerKA. New anticoagulants: Anti IIa vs anti Xa–is one better? J Thromb Thrombolysis, 2006; 21(1):67–72; doi: 10.1007/s11239-006-5579-4
49.
JauchEC, SaverJL, AdamsHPJr, et al.; Council on Clinical Cardiology. Guidelines for the early management of patients with acute ischemic stroke: A guideline for healthcare professionals from the American Heart Association/American Stroke Association. Stroke, 2013; 44(3):870–947; doi: 10.1161/STR.0b013e318284056a
50.
SimD, FlaumenhaftR, FurieB, et al.Interactions of platelets, blood-borne tissue factor, and fibrin during arteriolar thrombus formation in vivo. Microcirculation, 2005; 12(3):301–311; doi: 10.1080/10739680590925682
51.
PerzbornE, HeitmeierS, LauxV. Effects of Rivaroxaban on platelet activation and platelet-coagulation pathway interaction: In vitro and in vivo studies. J Cardiovasc Pharmacol Ther, 2015; 20(6):554–562; doi: 10.1177/1074248415578172
52.
OlivierCB, WeikP, MeyerM, et al.Dabigatran and rivaroxaban do not affect AA- and ADP-induced platelet aggregation in patients receiving concomitant platelet inhibitors. J Thromb Thrombolysis, 2016; 42(2):161–166; doi: 10.1007/s11239-016-1350-7
53.
JourdiG, Bachelot-LozaC, MazoyerE, et al.Effect of rivaroxaban and dabigatran on platelet functions: In vitro study. Thromb Res, 2019; 183:159–162; doi: 10.1016/j.thromres.2019.10.007
