Antiplatelet and anti-proliferative action of disintegrin from Echis multisquamatis snake venom
| Autor: | Liudmyla V. Garmanchuk, E. V. Lugovskoy, Platonova Tm, V. O. Chernyshenko, Chernyshenko Tm, Ludmila Kasatkina, O. V. Gornytska, O. I. Dzhus, D S Korolova, Natalia Petruk, Andriy Rebriev |
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| Rok vydání: | 2017 |
| Předmět: |
Blood Platelets
0301 basic medicine Platelet Aggregation Enzyme-Linked Immunosorbent Assay 030204 cardiovascular system & hematology Pharmacology complex mixtures HeLa 03 medical and health sciences 0302 clinical medicine Echis Viperidae biology.animal Disintegrin Animals Humans biology Cell growth Chemistry General Medicine Anti proliferative biology.organism_classification Molecular Weight 030104 developmental biology Snake venom biology.protein Platelet aggregation inhibitor Electrophoresis Polyacrylamide Gel Original Article Platelet Aggregation Inhibitors HeLa Cells Snake Venoms |
| Zdroj: | Croatian Medical Journal |
| ISSN: | 1332-8166 0353-9504 |
| Popis: | Aim To purify the platelet aggregation inhibitor from Echis multisquamatis snake venom (PAIEM) and characterize its effect on platelet aggregation and HeLa cell proliferation. Methods Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) were used for PAIEM identification. Platelet aggregation in the presence of PAIEM was studied on aggregometer Solar-AP2110. The changes of shape and granularity of platelets in the presence of PAIEM were studied on flow cytometer COULTER EPICS XL, and degranulation of platelets was estimated using spectrofluorimetry. Indirect enzyme-linked immunosorbent assay was used for the determination of target of PAIEM on platelet surface. An assay based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was used to evaluate the effect of PAIEM on the proliferation of HeLa cells in cell culture. Results The molecular weight of the protein purified from Echis multisquamatis venom was 14.9 kDa. Half-maximal inhibitory concentration (IC50) of PAIEM needed to inhibit adenosine diphosphate (ADP)-induced platelet aggregation was 7 μM. PAIEM did not affect thrombin- or ADP-induced platelet activation, but it did prevent binding of the anti-IIb antibody to glycoprotein IIb/IIIa (GPIIbIIIa)-receptor of adhered platelets and inhibited the viability of HeLa cells by 54%. Conclusion As a member of the disintegrin family, PAIEM inhibited platelet aggregation and cell proliferation possibly by blocking integrin-mediated interactions. However, it did not impair cellular signaling causing any changes in platelet shape and granularity and did not affect ADP-induced platelet degranulation. This disintegrin was shown to be a potent inhibitor of integrin-mediated cellular interactions including platelet aggregation or cancer cell proliferation. |
| Databáze: | OpenAIRE |
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