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.
Highlights
In snakebites cases, envenomations are associated with an amazing variety of pathophysiological manifestations, which differ among the various groups of snakes and even within the same genera and species [1]
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
The current study aimed to provide new information on the cytotoxicity mechanisms of Argentine Bothrops snake venoms on a skeletal muscle cell line
Summary
Envenomations are associated with an amazing variety of pathophysiological manifestations, which differ among the various groups of snakes and even within the same genera and species [1]. The majority of snakebites in northeastern Argentina are caused by B. alternatus (yarará grande) and B. diporus (yarará chica), reptiles that belong to the Viperidae family [3,4,5]. Their venoms induce predominantly local myotoxicity, characterized by myonecrosis often associated with other effects, such as hemorrhage, blistering and edema, in a complex pattern of local tissue damage [6, 7]. This cell culture is an appropriate in vitro model system to evaluate the myotoxicity and the events involved in the cell death caused by Bothrops venoms. In the current work we compared the toxic potency between both venoms on myoblast cell line and, in order to elucidate a putative direct cytotoxic action, we analysed the cytotoxicity and morphological features of cell death induced by B. alternatus and B. diporus venoms on the mouse myoblast continuous cell line, C2C12
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