Antithrombotic, Antiaggregant and Anticoagulant Effect of Methanolic Fraction of Calamintha officinalis: In vitro and ex vivo Experiments


  • University Mohammed the First, Laboratory of Physiology, Genetic and Ethnopharmacology, Faculty of Sciences, Oujda, 60000, Morocco
  • University Mohammed the First, Laboratory of Mineral Solid and Analytical Chemistry, Department of Chemistry, Faculty of Sciences, Oujda, 60000, Morocco
  • Laboratory Bensaid of Pathological Cytology and Anatomy, Oujda, 60000, Morocco


The objective was to study the effect of Calamintha officinalis methanolic fraction in the prevention of thrombosis and to explore its effect on primary and secondary hemostasis. The antithrombotic activity was evaluated by using the pulmonary thromboembolism experimental model. Platelet aggregation, tail bleeding time and coagulation assays were explored to elucidate the antithrombotic mechanism. For aggregation, washed platelets were incubated in presence of Calamintha officinalis methanolic fraction and then activated by different agonists (thrombin, ADP, collagen). The bleeding time was determined to measure the time between the tail incision and the moment the bleeding stops. The coagulation was explored by measuring prothrombin time, activated partial thromboplastin time, thrombin time and plasmatic fibrinogen concentration. For ex vivo study, the animals were treated by the Calamintha officinalis methanolic fraction (250 mg/kg/day) for 4 weeks. Then, platelet aggregation, tail bleeding time and coagulation parameters was studied. The results indicate that C. officinalis methanol fraction seems to exert in vivo an antithrombotic action (40% of protection). The in vitro investigations also show that this fraction significantly inhibits aggregation, increases bleeding time, coagulation times and reduces fibrinogen concentration. The chronic treatment of animals confirms the precedent data. This study demonstrates an antithrombotic effect of C. officinalis. The observed antiaggregant and anticoagulant effects could explain this activity.


Bleeding, Calamintha officinalis, Coagulation, Platelet Aggregation, Thrombosis.

Full Text:


El Haouari M, Rosado JA. Platelet function in hypertension. Blood Cells, Mol Dis. 2009; 42(1):38– 43. PMid: 18829351.

Nieswandt B, Pleines I, Bender M. Platelet adhesion and activation mechanisms in arterial thrombosis and ischaemic stroke. J Thromb Haemost. 2011; 9(1S):92–104.

Ringleb PA. Thrombolytics, anticoagulants, and antiplatelet agents. Stroke. 2006; 37(2):312–3. PMid: 16410469.

El-Hilaly J, Hmammouchi M, Lyoussi B. Ethnobotanical studies and economic evaluation of medicinal plants in Taounate province (Northern Morocco). J Ethnopharmacol. 2003; 86(2-3):149–58.

Merzouki A, Ed-derfoufi F, Molero Mesa J. Contribution to the knowledge of Rifian traditional medicine. II: Folk medicine in Ksar Lakbir district (NW Morocco). Fitoterapia. 2000; 71(3):278–307.

Monforte MT, TzakouO, Nostro A, Zimbalatti V, Galati EM. Chemical composition and biological activities of Calamintha officinalis Moench essential oil. J Med Food. 2011; 14(3):297–303. PMid: 21142949. https://

Singh PP, Jha S, Irchhaiya R. Antidiabetic and antioxidant activity of hydroxycinnamic acids from Calamintha officinalis Moench. Med Chem Research. 2012; 21(8):1717–21.

Lemhadri A, Zeggwagh NA, Maghrani M, Jouad, H, Michel JB, Eddouks M. Hypoglycaemic effect of Calamintha officinalis Moench in normal and streptozotocin-induced diabetic rats. J Pharm Pharmacol. 2004; 56:795–9. PMid: 15231045.

Di Minno G, Silver MJ. Mouse antithrombotic assay: a simple method for the evaluation of antithrombotic agents in vivo. Potentiation of antithrombotic activity by ethyl alcohol. J Pharmacol Exp Ther. 1983; 225(1):57–60.

Tomita T, Umegaki K, Hayashi E. Basic aggregation properties of washed rat platelets: Correlation between aggregation, phospholipid degradation, malondialdehyde and thromboxane formation. J Pharmacol Methods. 1983; 10(1):31–4.

Johansen M. Antiplatelet therapy after aspirininduced upper gastrointestinal bleeding. Tidsskr Nor Laegeforen. 2006; 126(2):2802–4. PMid: 17086221.

Asad M, Aslam M, Munir TA, Nadeem A. Effect of Acacia nilotica leaves extract on hyperglycaemia, lipid profile and platelet aggregation in streptozotocin induced diabetic rats. J Ayub Med Coll Abbottabad. 2011; 23(2):3–7. PMid: 24800330.

Janssen P, Mensink RP, Cox FJ, Harryvan JL, Hovenier R, Hollman PC, Katan MB. Effect of the flavonoids quercetin and apigenin on hemostasis in healthy voluters: Results from an in vitro and a dietary supplement study. Am J Clin Nutr. 1998; 67(2):255– 62. PMid: 9459373. ajcn/67.2.255.

Sikka P, Bindra VK. Newer antithrombotic drugs. Indian J Crit Care Med. 2010; 14(4):188–95. PMid: 21572750 PMCid: PMC3085220.

Son DJ, Cho MR, Jin YR, Kim SY, Park YH, Lee SH, Akiba S, Sato T, Yun YP. Antiplatelet effect of green tea catechins: a possible mechanism through arachidonic acid pathway. Prostaglandins Leukot Essent Fatty Acids. 2004; 71(1):25–31. PMid: 15172681.

Endale M, Lee WM, Kamruzzaman SM, Kim, SD, Park JY, Park MH, Rhee MH, Park TY, Park HJ, Cho JY, Rhee MH. Ginsenoside-Rp1 inhibits platelet activation and thrombus formation via impaired glycoprotein VI signalling pathway, tyrosine phosphorylation and MAPK activation. Br J Pharmacolo. 2012; 167(1):109–27. PMid: 22471932 PMCid: PMC3448917. j.1476-5381.2012.01967.x.

Dejana E, Villa S, de Gaetano G. Bleeding time in rats: A comparison of different experimental conditions. Thromb Haemost. 1982; 48(1):108–11.

Gadi D, Bnouham M, Aziz M, Ziyyat A, Legssyer A, Legrand C, Lafeve FF, Mekhfi H. Parsley extract inhibits in vitro and ex vivo platelet aggregation and prolongs bleeding time in rats. J Ethnopharmacol.

; 125(1):170–4. PMid:19467310.

Amirou A, Bnouham M, Legssyer A, Ziyyat A, Aziz M, Berrabah M, and Mekhfi H. Effects of Juglans regia root bark extract on platelet aggregation, bleeding time and plasmatic coagulation: in vitro and ex vivo Experiments. Evid Based Complementary Alternat Med. 2018; 7313517.

Mekhfi H, Gadi D, Bnouham M, Ziyyat A, Legssyer A, Aziz M. Effect of argan oil on platelet aggregation and bleeding time: A beneficial nutritional property. J Complement Integr Med. 2008; 5(1):18.

Kamruzzaman SM, Endale M, Oh WJ, Park SC, Kim KS, Hong JH, Kwak YS, Yun BS, Rhee MH. Inhibitory effects of Bulnesia sarmienti aqueous extract on agonist-induced platelet activation and thrombus formation involves mitogen-activated protein kinases. J Ethnopharmacol. 2010; 130(3):614– 20. PMid: 20558266.

Renne T, Nieswandt B, Gailani D. The intrinsic pathway of coagulation is essential for thrombus stability in mice. Blood Cells, Mol Dis. 2006; 36(2):148–51. PMid: 16466946.

Schulman S, Bijsterveld NR. Anticoagulants and their reversal. Transfus Med Rev. 2007; 21(1):37– 48. PMid: 17174219.

Li H, Huang W, Wen Y, Gong G, Zhao Q, Yu G. Antithrombotic activity and chemical characterization of steroidal saponins from Dioscorea zingiberensis C.H. Wright. Fitoterapia. 2010; 81(8):1147–56. PMid: 20659537.

Wang Y, Shao J, Yao S, Zhang S, Yan J, Wang H, Chen Y. Study on the antithrombotic activity of Umbilicaria esculenta polysaccharide. Carbohydr Polym. 2014; 105(1):231–6. PMid: 24708975.

Singh PP, Jha S, Irchhaiya R. Pharmacognostical and physicochemical investigation of the leaf of Calamintha officinalis moench. Asian Pac J Trop Biomed. 2012; 2(3S):S1362–6.

Lee W, Lee J, Kulkarni R, Kim MA, Hwang JS, Na M, Bae JS. Antithrombotic and antiplatelet activities of small-molecule alkaloids from Scolopendra subspinipes mutilans. Sci Rep. 2016; 6(1):21956. PMid: 26905699 PMCid: PMC4764974.

Mekhfi H, El Haouari M, Bnouham M, Aziz M, Ziyyat A, Legssyer A. Effects of extracts and tannins from Arbutus unedo leaves on rat platelet aggregation. Phytother Res. 2006; 20(2):135–9. PMid: 16444667.

El Haouari M, Mekhfi H. Antiplatelet aggregation effects of extracts from Arbutus unedo leaves. Plant Science Today. 2017; 4(2):68–74.

Quiñones M, Miguel M, Aleixandre A. Beneficial effects of polyphenols on cardiovascular disease. Pharmacol Res. 2013; 68(1):125–31. PMid:23174266.

Middleton E Jr, Kandaswami C, Theoharides TC. The effects of plant flavonoids on mammalian cells: Implications for inflammation, heart disease and cancer. Pharmacol Rev. 2000; 52(4):673–751. PMid: 11121513.

Kundu JK, Shin YK, Kim SH, Surh YJ. Resveratrol inhibits phorbol ester-induced expression of COX-2 and activation of NF-kB in mouse skin by blocking IkB kinase activity. Carcinogenesis. 2006; 27(7):1465–74. PMid: 16474181.

Manna SK, Mukhopadhyay A, Aggarwal BB. Resveratrol suppresses TNF-induced activation of nuclear transcription factors NF- B, activator protein-1 and apoptosis: Potential Role of Reactive Oxygen Intermediates and Lipid Peroxidation. J Immunol. 2000; 164(12):6509–19. PMid: 10843709.

Monforte MT, Lanuzza F, Pergolizzi S, Mondello F, Tzakou O, Galati EM. Protective effect of Calamintha officinalis moench leaves against alcohol-induced gastric mucosa injury in rats. Macroscopic, histologic and phytochemical analysis. Phytother Res. 2012; 26(6):839–44. PMid: 22076933.

Lee DH, Kim HH, Cho HJ, Bae JS, Yu YB, Park HJ. Antiplatelet effects of caffeic acid due to Ca2+ mobilization inhibition via cAMPdependent inositol-1, 4, 5-trisphosphate receptor phosphorylation. J Atheroscler Thrombosis. 2014; 21(1):23–37. PMid:24088646.

Luceri C, Giannini L, Lodovici M, Antonucci E, Abbate R, Masini E, Dolara P. p-Coumaric acid, a common dietary phenol, inhibits platelet activity in vitro and in vivo. Br J Nutr. 2007; 97(3):458–63. PMid: 17313706.

Fuentes E, Caballero J, Alarcon M, Rojas A, Palomo I. Chlorogenic acid inhibits human platelet activation and thrombus formation. PloS one. 2014; 9(3):e90699, 1–13.


  • There are currently no refbacks.