ACTION OF VENOM OF VIPERA LEBETINA ON BLOOD COAGULATION in vitro
نویسندگان
چکیده
Aim. In this study we focused on the search of fibrinogen-targeted proteases in venom Vipera lebetina. Methods. Fractionation was performed using FPLC chromatographic system Acta Prime Q Sepharose. Analysis protein mixtures SDS-PAGE. Аction blood coagulation analyzed APTT assay [2]. Proteolytic action fibrinogen and identification components with fibrinolytic activity electrophoresis mixture solution (2 mg/ml) venom`s fractions. For a comprehensive evaluation effect obtained fractions hemostasis, an original approach modified aggregatometry used [3]. This made it possible to take into account ability activate platelets, initiate coagulation, or inhibit platelet aggregation. Hemolytic estimated fresh human red cells. Amount released hemoglobin by spectrophotometry Optizen POP. Results. Crude V. lebetina fractionated ion-exchange chromatography Elution stepwise gradient NaCl (0.1, 0.2, 0.3, 0.5, 0.7 M NaCl) 0.05 Tris-HCl buffer at pH 8.3. Fractions eluted 0.1 0.2 contained several proteins different molecular weights ranging from 75 kDa low weight according Proteins that cleave α- β-chains were found indicating presence enzyme fibrinogenolytic The did not show any significant activity. After aggregation concluded fraction An increase observed for after addition ADP. may indicate active compound promotes Further research is necessary determine its nature. had no A decrease plasma clotting time 5 s 7 s, compared control value 70 shown concentrations M, respectively. 0.5 only slight reducing clotting. slightly increased level hemolysis unbound whole venom. It can be suggested phospholipase are present non-binded fraction. Conclusions. Thus, fibrinogen-specific proteases, hemolytic agents, activators Most these compounds must purified basic biochemical research.
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ژورنال
عنوان ژورنال: Biotechnologia Acta
سال: 2023
ISSN: ['2410-776X', '2410-7751']
DOI: https://doi.org/10.15407/biotech16.02.024