Cytotoxic and inflammatory potential of a phospholipase A2 from Bothrops jararaca snake venom

Abstract

BackgroundSnake venom phospholipases A2 (PLA2s) have been reported to induce myotoxic, neurotoxic, hemolytic, edematogenic, cytotoxic and proinflammatory effects. This work aimed at the isolation and functional characterization of a PLA2 isolated from Bothrops jararaca venom, named BJ-PLA2-I.Methods and ResultsFor its purification, three consecutive chromatographic steps were used (Sephacryl S-200, Source 15Q and Mono Q 5/50 GL). BJ-PLA2-I showed acidic characteristics, with pI~ 4.4 and molecular mass of 14.2 kDa. Sequencing resulted in 60 amino acid residues that showed high similarity to other Bothrops PLA2s, including 100% identity with BJ-PLA2, an Asp49 PLA2 previously isolated from B. jararaca venom. Being an Asp49 PLA2, BJ-PLA2-I showed high catalytic activity, and also inhibitory effects on the ADP-induced platelet aggregation. Its inflammatory characterization showed that BJ-PLA2-I was able to promote leukocyte migration in mice at different concentrations (5, 10 and 20 μg/mL) and also at different response periods (2, 4 and 24 h), mainly by stimulating neutrophil infiltration. Furthermore, increased levels of total proteins, IL-6, IL-1β and PGE2 were observed in the inflammatory exudate induced by BJ-PLA2-I, while nitric oxide, TNF-α, IL-10 and LTB4 levels were not significantly altered. This toxin was also evaluated for its cytotoxic potential on normal (PBMC) and tumor cell lines (HL-60 and HepG2). Overall, BJ-PLA2-I (2.5–160 μg/mL) promoted low cytotoxicity, with cell viabilities mostly varying between 70 and 80% and significant values obtained for HL-60 and PBMC only at the highest concentrations of the toxin evaluated.ConclusionsBJ-PLA2-I was characterized as an acidic Asp49 PLA2 that induces acute local inflammation and low cytotoxicity. These results should contribute to elucidate the action mechanisms of snake venom PLA2s.

Effect of Bitis gabonica and Dendroaspis angusticeps snake venoms on apoptosis-related genes in human thymic epithelial cells

“…Boeno et al [ 90 ] have shown that purified Lys49-PLA2 homologue from Bothrops jararacussu might contribute to inflammasome formation through the activation of NLRP3, which in turn interacts with caspase-1, leading to an inflammatory response via IL-1β. PLA2 mediated inflammatory response was observed by Cedro et al [ 91 ]. Using purified Asp49-PLA2 (5, 10 and 20 μg/mL) from Bothrops jararaca snake venom, an increased level of IL-6, IL-1β and PGE2 was described through neutrophil stimulation, although its cytotoxic activity is inferior to those observed in case of the Lys49-PLA2 homologue structure.…”

Antibiofilm Activity of Acidic Phospholipase Isoform Isolated from Bothrops erythromelas Snake Venom

“…Other reports show the use of up to three stages, such as studies involving the species Bothrops atrox and Bothrops jararaca, which were subjected to different stages and buffers. The other purifications mentioned were the use of a C18 column in the RP-HPLC stage, differing only from BE-I-PLA 2 isolated from B. erythromelas [34,38,39].…”

Bioassay-guided fractionation, phospholipase A2-inhibitory activity and structure elucidation of compounds from leaves of Schumanniophyton magnificum

“…PLA 2 hydrolyses fatty acids at the sn-2 position of membrane phospholipids releasing lysophospholipids and arachidonic acid (Cedro et al 2018). Arachidonic acid in turn leads to the production of eicosanoids such as prostaglandins, thromboxanes and leukotrienes (Yui et al 2015), which are mediators of inflammatory effect.…”