Oxidative stress induced by crude venom from the jellyfish Pelagia noctiluca in neuronal-like differentiated SH-SY5Y cells.
ABSTRACT Marine toxins are a suitable research model and their mechanism of action is intriguing and still under debate. Either a pore formation mechanism or oxidative stress phenomena may explain the damage induced by toxins. The effect of crude venom from isolated nematocysts of the jellyfish Pelagia noctiluca on neuronal-like cells derived from human neuroblastoma SH-SY5Y has been here studied. To prove the possible oxidative stress events, cell viability, assessed by MTT quantitative colorimetric assay, intracellular reactive oxygen species (ROS) quantified by the non-fluorescent probe H2DCF-DA and changes in mitochondrial transmembrane potential (ΔΨm) measured by the incorporation of a cationic fluorescent dye rhodamine-123 were verified on venom-treated cells (0.05-0.5μg/ml doses). A dose- and time-dependent reduction of all parameters was observed after venom treatment. NAC (N-acetyl-cysteine), antioxidant applied before crude venom application, significantly counteracted the decrease in cell viability and ROS production, while ΔΨm was only partially restored. The disruption of mitochondrial membrane by P. noctiluca crude venom may thus induce oxidative stress by inhibiting mitochondrial respiration and uncoupling oxidative phosphorylation, sensitizing mitochondria in SH-SY5H cells and facilitating membrane permeability. In sum, our findings suggest that P. noctiluca crude venom directly induces ΔΨm collapse with further generation of ROS and add novel information to the understanding of such toxins, still not completely clarified.
- SourceAvailable from: Giulio KleinerCell Death & Disease 01/2013; 4:e585. · 6.04 Impact Factor
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ABSTRACT: The cytotoxic effects of the crown-ofthorns starfish Acanthaster planci spine venom (ASV) in five cell lines, including human neuroblastoma (SHSY5Y), human hepatocellular carcinoma (HepG2), human melanoma (A375.S2), human skin fibroblast (CCD-966SK) and mouse macrophage-like cell (RAW 264.7) were assayed. The results indicated that ASV showed cytotoxic effects depending on dose in these five cell lines. Specifically, ASV significantly inhibited the proliferation of human melanoma cell line A375.S2 at 10 μg/mL, indicating A. planci spine venom could be utilized as potential chemotherapeutic agent in the treatment of cancer. The cytotoxicity of ASV to human melanoma cell line A375.S2 was relatively well retained at temperature less than 40°C, and sharply lost at temperature more than 80°C. The cytotoxicity of ASV also sharply lost at extreme pH environments (pH 2 and 12). The cytotoxicity of ASV was attenuated when treated with Cu2+ and anti-oxidant N-acetylcysteine. After SDS-PAGE analysis, ASV showed the major protein components ranging from 10 kDa to 37 kDa.Molecular and Cellular Toxicology 9(2). · 0.72 Impact Factor