Amidolytic thrombin activity under the ischemic stroke peptide pool influence

T. Katrii, O. Savchuk (Kiev, Ukraine)

Bleeding disorders, coagulation and fibrinolytic factors
Date: 17.02.2017,
Time: 17:15 - 18:15

Objective: Group of peptides 3-5 kDa are able to reflect the level of metabolic disorders in the organism (3). Clear and unambiguous answer to the question regarding protein pool (PP) properties is absent. At the same time, stroke is one of the most pressing health problem which require urgent solution (4). Full recovery of patients after stroke was not observed (5). The leading mechanism of ischemic stroke realization correlated with haemostatic profile (6-8). And the thrombin is key factor of haemostasis (1,2). Investigation of the potential influence of PP formed in the bloodstream after suffering a stroke, on the amidolytic activity of thrombin has utmost importance.

Methods: PP fractions were obtained by the NikolaichykV.(9) method from the blood plasma of 35 healthy donors and 56 atherothrombotic (AIS) and cardioembolic (CIS) ischemic stroke patients in acute phase as well as 56 patients one year past acute phase. Isolated PP fractions were dialyzed against vehicle. Experimental mixture preparation and registration of absorption were done like previously (10). An activator of prothrombin derived from the venom Echis multisquamatus (ecamylin) were used (11). Control sample contained the same components but equal volume of vehicle instead of PP. Different concentrated PP fractions were tested.

Results: Different concentrations of PP showed various and often opposite effects. Thus PP fraction derived from blood plasma of healthy donors inhibited tested activity of thrombin in concentrations lower then avarege PP concentration analogical group. The maximum inhibition wes observed under influence of healthy donors PP fractions in concentration half (34 mkg/ml) and quote (17 mkg/ml) less than average concentration in analogical group by the 12% and 20% correspondingly. In contrast stroke PP fractions led to opposite effects. Maximum effect of PP fractions on amidolytic thrombin activity was observed by using half less concentrated PP. Thus PP from acute stroke AIS as well as CIS activated study process by 20% and 8% respectively. Oposite effect observed for thrombin activation from the prothrombin in plasma.

Conclusion: Therefore results affirm that ischemic stroke accompanied by the formation of the peptide pool in the bloodstream that could take part in recurrence of the disease. 1.      Mann KG. (2003) Thrombin formation. Chest. 124(3):4S-10S. 2.      Wolberg A, Campbell R. (2008) Thrombin Generation, Fibrin Clot Formation and Hemostasis. Transfus Apher Sci. 38(1): 15–23. 3.      Karjakin E.V, Belov S.V. (2004) The peptide pool as integral indicator of metabolic disorders. Biochemistry.3-8. 4.      Hoyert D, Land Xu. (2012) National vital statistics reports: from the Centers for Disease Control and Prevention, National Center for Health Statistics, National Vital Statistics System. 61.6. 1-52. 5.      Shuaib A, Hussain M. (2008) The past future of neuroprotection in cerebral ischemic stroke. Eur. Neurol. 59. 41-43. 6.      Woodward Ì, Lowe D, Camðbell J. (2005) Associations of inflammatory and hemostatic variables with the risk of recurrent stroke. Stroke. 36. 2143–2147. 7.       Hirsh Jack. (2006) Hemostasis and thrombosis: basic principles and clinical practice. 8.      Martin J, Shaw T, Heggie J. (1983) The biological significance of platelet volume: its relationship to bleeding time, platelet thromboxane B 2 production and megakaryocyte nuclear DNA concentration. Thrombosis research. 32.5. 443-460. 9.      Nykolaychyk B. B Moyn VM, Kyrkovskyy VV (1991) Method for determining of the peptide pool moleculars. Laboratornoe case. 10. 13-18. 10.  Katrii TB, Kravchenko NK, Vovk TB, Ostapchenko LI, Savchuk OM. (2016) The influence of immunoglobulin class G from blood plasma of patients with stroke on the activity of some parameters of  haemostasis system. Jornal Blood Coagulation&Fibrinolisis. Jen; Doi: 10.109/MBC.0000000000000497. 11.  Volkov G., Savchuk A., Karbovskyy V. (2008) Method of extraction protein C activator from Agristrodon blomhoffii ussuriensis venom. Patent WO 2008/020740.