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


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Article Info

DATE

2018-11-23

AUTHORS

Rafhaella C. A. Cedro, Danilo L. Menaldo, Tássia R. Costa, Karina F. Zoccal, Marco A. Sartim, Norival A. Santos-Filho, Lúcia H. Faccioli, Suely V. Sampaio

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. More... »

PAGES

33

References to SciGraph publications

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s40409-018-0170-y

    DOI

    http://dx.doi.org/10.1186/s40409-018-0170-y

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1110157122

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/30498509


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    16 schema:description 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.
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    23 ADP
    24 B. jararaca venom
    25 Bothrops jararaca snake venom
    26 Bothrops jararaca venom
    27 HL-60
    28 IL-10
    29 IL-1β
    30 IL-6
    31 LTB4 levels
    32 PBMC
    33 PGE2
    34 PLA2
    35 ResultsFor
    36 TNF
    37 acid residues
    38 acidic characteristics
    39 action mechanism
    40 activity
    41 acute local inflammation
    42 aggregation
    43 amino acid residues
    44 catalytic activity
    45 cell lines
    46 cell viability
    47 characteristics
    48 characterization
    49 chromatographic steps
    50 concentration
    51 consecutive chromatographic steps
    52 cytotoxic
    53 cytotoxic potential
    54 cytotoxicity
    55 different concentrations
    56 effect
    57 exudates
    58 functional characterization
    59 high catalytic activity
    60 high concentrations
    61 high similarity
    62 identity
    63 infiltration
    64 inflammation
    65 inflammatory characterization
    66 inflammatory exudate
    67 inflammatory potential
    68 inhibitory effect
    69 isolation
    70 jararaca venom
    71 leukocyte migration
    72 levels
    73 lines
    74 local inflammation
    75 low cytotoxicity
    76 mass
    77 mechanism
    78 method
    79 mice
    80 migration
    81 molecular mass
    82 myotoxic
    83 neutrophil infiltration
    84 nitric oxide
    85 oxide
    86 period
    87 phospholipase A2
    88 platelet aggregation
    89 potential
    90 proinflammatory effects
    91 protein
    92 purification
    93 residues
    94 response period
    95 results
    96 sequencing
    97 significant value
    98 similarity
    99 snake venom
    100 snake venom PLA2
    101 step
    102 total protein
    103 toxin
    104 tumor cell lines
    105 values
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    108 viability
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