Cytotoxicity and morphological analysis of cell death induced by Bothrops venoms from the northeast of Argentina

Abstract

Bothrops snake venoms have been proved toxic to a variety of cell types, in both in vivo and in vitro models. Studies on the pharmacological actions of Bothrops venoms from Argentina are relatively scarce and the direct action of the crude venoms has not been assessed using cell culture models. In this work, we investigated the cytotoxicity of crude venoms from B. alternatus and B. diporus in a skeletal muscle (C2C12) cell line, which is commonly used as a model for studying the myotoxic action of snake venom. Both venoms (1.25-50 µg/mL) induced an early and significant decrease in cell viability. The cytotoxic concentration 50 (CC50), determined three hours after exposure, revealed that B. diporus venom was significantly more cytotoxic (CC50: 2 µg/mL) than B. alternatus (CC50: 5.8 µg/mL). To investigate the cell death mechanism involved, myoblast cells were examined by phase contrast microscopy and after acridine orange and ethidium bromide fluorescence staining, respectively. Our data clearly demonstrated that an apoptotic mechanism mediated this cell line destruction. The current study aimed to provide new information on the cytotoxicity mechanisms of Argentine Bothrops snake venoms on a skeletal muscle cell line

Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

“…Our interpretation of the results suggests that the LD 50 value is approximately ten times greater than the IC 50 value for crude venom from Macrovipera lebetina lebetina. Our results agree with those from other authors (40)(41)(42). Another important point is that the venom enhances the cytotoxic response of L929 cells when administered for longer periods (48 hours) with an IC 50 value of 0.62 ± 0.18 µg/ mL.…”

“…Currently, researchers suggest that cell-based assay to examine venom cytotoxicity is an alternative to animal testing (18,19,(40)(41)(42). Our in vitro results show that crude venom from Macrovipera lebetina lebetina is highly cytotoxic for cultured fibroblasts causing decreased viability, the disappearance of normal morphological characteristics, rounding up, detachment and death at the highest concentration.…”

Screening of cytotoxic and antimicrobial activity potential of AnatolianMacrovipera lebetina obtusa(Ophidia: Viperidae) crude venom

“…With regard to the cytotoxicity results of this study, the crude venom of M. l. obtusa showed dose-dependent cytotoxic effects at various levels on some of the selected cell lines, corroborating previous reports which show cytotoxic effects of different crude snake venoms or purified venomic proteins/peptides on various cancer and non-cancerous cells (Bustillo et al 2009;Jamunaa et al 2012;Yalcin et al 2014). Under the experimental conditions in this study, M. l. obtusa venom was more potent against Vero, U-87 MG, MCF-7 and CaCo-2 cell lines, compared to HeLa and A549 cells.…”

VIABILITY OF VERO E6 CELLS EXPOSED TO THE Bothrops leucurus SNAKE CRUDE VENOM

“…The study using a murine skeletal muscle cell line revealed that an apoptotic mechanism mediates the cellular destruction caused by Bothrops venoms (BUSTILLO et al, 2009 (NUNES et al, 2012). In a study using ISSN: 2318-356X V.6 N.3 SET/DEZ 2018 BlV on MDCK cells, cell death was induced predominantly by necrosis, with presence of pycnotic nuclei, increase in [Ca 2+ ] cyt , and decrease of membrane potential in mitochondria (MORAIS, 2011).…”

“…InBustillo et al's (2009) study, it was verified that after incubation of C2C12 murine cells with 10 μg/mL of B. alternatus venom (BaV), several morphological alterations occurred, as reduction of nuclear sizes, chromatin condensation, cell rounding, detachment cells, cell shrinkage and the formation of blebs on cell surface.Venoms contain many toxic components and can target different cell types, resulting in a multiplicity of pathological lesions. The detachment activity of the monolayer can be explained by the presence of desintegrins derived from snake venom, whose effect would be to block the integrins, membrane surface proteins responsible for cell differentiation, proliferation and activation ROJNUCKARIN, 2008).…”