Antiplatelet and anti-proliferative action of disintegrin from Echis multisquamatis snake venom

Abstract

AimTo purify the platelet aggregation inhibitor from Echis multisquamatis snake venom (PAIEM) and characterize its effect on platelet aggregation and HeLa cell proliferation.MethodsSodium 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.ResultsThe 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%.ConclusionAs 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.

Cytotoxic and anticancer properties of the Malaysian mangrove pit viper (Trimeresurus purpureomaculatus) venom and its disintegrin (purpureomaculin)

“…This was a purified protein validated through SDS-PAGE and LC-MS/MS, and termed purpureomaculin in line with the naming of most snake venom-derived disintegrin protein (prefix "species-" + post-fix "-in"). The finding of purpureomaculin anticancer activity is consistent with earlier studies that reported the inhibitory activity of snake venom disintegrins on the proliferation, metastasis and adhesion of cancer cells [21,25,37]. The selective cytotoxic effect of purpureomaculin in breast cancer cells (MCF7) corroborates the selective cytotoxicity of MTP venom in the same cancer cell line observed in this study.…”

“…The antineoplastic potential of snake venom disintegrins have been shown in several studies, e.g. contortrostatin (disintegrin isolated from Agkistrodon contortrix venom) [23], saxatilin (disintegrin from Gloydius saxatilis venom) [24], PAIEM (disintegrin isolated from Echis multisquamatis venom) [25] and a disintegrin from Crotalus durissus collilineatus venom [26]. The disintegrins act by inhibiting angiogenesis, invasion and migration of cancer cells.…”

Preparation of highly-concentrated autologous platelet-rich plasma for biomedical use

“…This method is easy to use and does not require expensive devices. It was successfully used in our laboratory and allowed us to obtain data with high reproducibility [16,17].…”

Bibliometric Analysis of Literature in Snake Venom-Related Research Worldwide (1933–2022)

“…Concerns over the swift rise in microbial drug resistance have prompted researchers to explore the promising role of snake venom and its components in eradicating superbugs ( Table S1 ) [ 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 ]. In parallel, numerous studies have also been conducted to isolate and characterize peptides found in snake venoms as potential cancer drugs ( Table S2 ) [ 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 ]. Additionally, in recent years, conjugation of s...…”