Cardiovascular and biochemical studies on the effects of thrombin and dabigatran and the interaction with vasopressor molecules
Keywords:
Thrombin, Dabigatran, Phospholipase C, Blood pressure, Heart rateAbstract
Background: The effect of serine protease thrombin and its directly acting inhibitor dabigatran were evaluated on the heart rate, blood pressure, and phospholipase C (PLC) enzyme activity and the intracellular calcium levels in the platelets.
Methods: Heart rate and blood pressure were estimated using electrophysiology equipment.
Results: While thrombin was unable to significantly affect the heart rate and blood pressure, the inhibitor dabigatran was able to reduce the heart rate appreciably but its effects on the blood pressure were minimal. The thrombin induced increase in PLC enzyme activity, and intracellular calcium levels were attenuated by dabigatran in the platelets. The posterior pituitary hormone, vasopressin, and the adrenergic agonist noradrenaline were used to stimulate the PLC and calcium levels in platelets.
Conclusion: The thrombin inhibitor, dabigatran reduces vascular oxidative stress and inflammation, improves endothelial function, and decreases atherosclerosis in rodents.
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References
Kataoka H, Hamilton JR, McKemy DD, Camerer E, Zheng YW, Cheng A, et al. Protease-activated receptors 1 and 4 mediate thrombin signaling in endothelial cells. Blood. 2003;102(9):3224-31.
Mizuno O, Hirano K, Nishimura J, Kubo C, Kanaide H. Mechanism of endothelium-dependent relaxation induced by thrombin in the pig coronary artery. Eur J Pharmacol. 1998;351(1):67-77.
Hamilton JR, Cocks TM. Heterogeneous mechanisms of endothelium-dependent relaxation for thrombin and peptide activators of protease-activated receptor-1 in porcine isolated coronary artery. Br J Pharmacol. 2000;130(1):181-8.
Hirano K. The roles of proteinase-activated receptors in the vascular physiology and pathophysiology. Arterioscler Thromb Vasc Biol 2007;279(1):27-36.
Baetz BE, Spinler SA. Dabigatran etexilate: an oral direct thrombin inhibitor for prophylaxis and treatment of thromboembolic diseases. Pharmacotherapy. 2008;28(11):1354-73.
Eisert WG, Hauel N, Stangier J, Wienen W, Clemens A, van Ryn J. Dabigatran: an oral novel potent reversible nonpeptide inhibitor of thrombin. Arterioscler Thromb Vasc Biol. 2010;30(10):1885-9.
O’Rourke FA, Halenda SP, Zavoico GB, Feinstein MB. Inositol 1,4,5-trisphosphate releases Ca2+ from a Ca2+-transporting membrane vesicle fraction derived from human platelets. J Biol Chem. 1985;260(2):956-62.
Hathaway DR, Adelstein RS. Human platelet myosin light chain kinase requires the calcium-binding protein calmodulin for activity. Proc Natl Acad Sci U S A. 1979;76(4):1653-7.
Barrios V, Escobar C. Can dabigatran improve blood pressure control? Future Cardiol. 2013;9(3):321-3.
Clemens A, Fraessdorf M, Friedman J. Cardiovascular outcomes during treatment with dabigatran: comprehensive analysis of individual subject data by treatment. Vasc Health Risk Manag. 2013;9:599-615.
Parasuraman S, Raveendran R. Measurement of invasive blood pressure in rats. J Pharmacol Pharmacother. 2012;3(2):172-7.
Tyagi MG, Thomas M. Enhanced cardiovascular reactivity to desmopressin in water-restricted rats: facilitatory role of immunosuppression. Methods Find Exp Clin Pharmacol. 1999;21(9):619-24.
Deepika DV, Bhavapriya R, Ramaswamy A, Tyagi MG. Influence of PI-3 kinase inhibition and plasminogen activation on phospholipase C and D enzyme activity in goat kidney. Int J Biotech Biochem. 2013;9(3):341-9.
Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985;260(6):3440-50.
Chang CJ, Chen YC, Kao YH, Lin YK, Chen SA, Chen YJ. Dabigatran and thrombin modulate electrophysiological characteristics of pulmonary vein and left atrium. Circ Arrhythm Electrophysiol. 2012;5(6):1176-83.
Connolly SJ, Ezekowitz MD, Yusuf S, Eikelboom J, Oldgren J, Parekh A, et al. Dabigatran versus warfarin in patients with atrial fibrillation. N Engl J Med. 2009;361(12):1139-51.
Eriksson BI, Quinlan DJ, Weitz JI. Comparative pharmacodynamics and pharmacokinetics of oral direct thrombin and factor Xa inhibitors in development. Clin Pharmacokinet. 2009;48(1):1-22.
Weitz JI, Leslie B, Hudoba M. Thrombin binds to soluble fibrin degradation products where it is protected from inhibition by heparin-antithrombin but susceptible to inactivation by antithrombin-independent inhibitors. Circulation. 1998;97(6):544-52.
Ruggeri ZM. Platelets in atherothrombosis. Nat Med. 2002;8(11):1227-34.
Billah MM, Lapetina EG. Rapid decrease of phosphatidylinositol 4,5-bisphosphate in thrombin-stimulated platelets. J Biol Chem. 1982;257(21):12705-8.
Kadamur G, Ross EM. Mammalian phospholipase C. Annu Rev Physiol. 2013;75:127-54.
Agranoff BW, Murthy P, Seguin EB. Thrombin-induced phosphodiesteratic cleavage of phosphatidylinositol bisphosphate in human platelets. J Biol Chem. 1983;258(4):2076-8.
Garcia JG, Patterson C, Bahler C, Aschner J, Hart CM, English D. Thrombin receptor activating peptides induce Ca2+ mobilization, barrier dysfunction, prostaglandin synthesis, and platelet-derived growth factor mRNA expression in cultured endothelium. J Cell Physiol. 1993;156(3):541-9.
Riewald M, Ruf W. Mechanistic coupling of protease signaling and initiation of coagulation by tissue factor. Proc Natl Acad Sci U S A. 2001;98(14):7742-7.
Hattori R, Hamilton KK, Fugate RD, McEver RP, Sims PJ. Stimulated secretion of endothelial von Willebrand factor is accompanied by rapid redistribution to the cell surface of the intracellular granule membrane protein GMP-140. J Biol Chem. 1989;264(14):7768-71.
André P, Denis CV, Ware J, Saffaripour S, Hynes RO, Ruggeri ZM, et al. Platelets adhere to and translocate on von Willebrand factor presented by endothelium in stimulated veins. Blood. 2000;96(10):3322-8.
Kroll MH, Schafer AI. Biochemical mechanisms of platelet activation. Blood. 1989;74(4):1181-95.
Vouret-Craviari V, Bourcier C, Boulter E, van Obberghen-Schilling E. Distinct signals via Rho GTPases and Src drive shape changes by thrombin and sphingosine-1-phosphate in endothelial cells. J Cell Sci. 2002;115:2475-84.
Michel MC, Brass LF, Williams A, Bokoch GM, LaMorte VJ, Motulsky HJ. Alpha 2-adrenergic receptor stimulation mobilizes intracellular Ca2+ in human erythroleukemia cells. J Biol Chem. 1989;264(9):4986-91.
Imai A, Nakashima S, Nozawa Y. The rapid polyphosphoinositide metabolism may be a triggering event for thrombin-mediated stimulation of human platelets. Biochem Biophys Res Commun. 1983;110:108-15.