ALT-C, a disintegrin-like Cys-rich protein from Bothrops alternatus, increases skeletal myoblast viability

Abstract

ALT-C, an ECD motif (glutamic acid, cysteine, aspartic acid) disintegrin from Bothrops alternatus snake venom, induces α2β1 integrin-mediated signaling and neutrophil chemotaxis. In vitro, in human umbilical vein endothelial cells (HUVEC), ALT-C induces cell proliferation, thus showing an interesting potential for tissue regeneration studies. This work aimed to evaluate the influence of ALT-C in myoblast viability and differentiation. Myoblasts were obtained from hind limb muscles of 3 to 4-day old Wistar rats. The cells were incubated with ALT-C at different concentrations and incubation periods were followed by total RNA isolation. cDNA synthesis and real time polymerase chain reaction (PCR) were performed with primers of myoD as well as of both (slow and fast) myosin heavy chain isoforms (MHC). ECD-disintegrin increased myoblast viability in a dose-dependent way, mostly with 50 to 100 nM concentrations, and such effect was more prevalent after 48 hours. No changes in gene expression of both MHC isoforms were observed in ALT-C-treated cells. MyoD expression was not detected, which suggests that myoblasts were in mature stages. Protease activity and cytokine array tested in a medium of 50 nM ALT-C-treated cells after 48 hours were not different from controls. In conclusion, it was shown that myoblats are sensitive to ALT-C indicating an integrin-mediated intracellular signaling that increases cell viability.

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

“…The venom increased the viability of HeLa cells at lower doses (1.25 and 2.5 μg/ml). This effect can be observed for snake venoms and also scorpion venoms because of proteins, peptides and other organic compounds that may promote cell proliferation at lower concentrations (Mesquita-Ferrari et al 2009;Erdeş et al 2014;Yalcin et al 2014). According to the results, HeLa (IC 50 = 12.80 μg/ml) is the most resistant cell line against all venom doses, while Vero and U-87 MG cell lines seem to be the most sensitive of the tested cell lines.…”

Rat skin wound healing induced by alternagin‐C, a disintegrin‐like, Cys‐rich protein from Bothrops alternatus venom

“…Previously results from our group have shown that ALT‐C strongly induces human vein endothelial cell (HUVEC) proliferation both in vitro and in vivo by up‐regulating the expression of some growth factors and their receptors, including vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR‐2) (18,19). In addition, it was shown that ALT‐C increases myoblast proliferation, modifies gene expression including the pattern of MHC gene expression (20) and modulates matrix metalloprotease (MMP) activity and expression in an in vivo skeletal muscle regeneration model (21).…”

A novel lipocalin homologue from the venom gland of Deinagkistrodon acutus similar to mammalian lipocalins

“…Due to frequent venom collection, inflammatory diseases in snake venom glands often occur and the pathogens are usually identified as bacteria. Although purification and gene expression of snake venom components have been extensively studied, there is no report on lipocalin homologue in snake venom glands (18)(19)(20)(21). Herein, the molecular cloning of a fulllength cDNA encoding a lipocalin homologue (d-lipo) in Deinagkistrodon acutus venom gland is reported, as well as the phylogenetic relationships among d-lipo and known homologues from other species.…”