VIABILITY OF VERO E6 CELLS EXPOSED TO THE Bothrops leucurus SNAKE CRUDE VENOM

J. Braga, Bruna Dos Santos Bispo, Victor Lucas Dias Cavalcante, S. Sardi, G. Campos, I. Biondi, Luis Eduardo Meira Faria
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Abstract

Primary and continuous cell culture systems are a good in vitro alternative for the quantitative assessment of snake venom toxicity. There are still few studies evaluating the cellular response against Bothrops leucurus venom (BlV). This study aimed to evaluate the cytotoxic effect of crude BlV on culture of VERO E6 cells. Neutral Red incorporation (NR) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) tests were performed for viable cells, and Trypan Blue (TB) dye exclusion test was performed for non-viable cells. The differences between the groups were evaluated by ANOVA-One-Way and Dunnet through the program Graph Pad 5.0 with significance of 5%. The 50% cytotoxic concentration (CC50) of BlV was 7.86 μg/mL. There was a trend of dose-dependent reduction on cell viability in all assays evaluated. However, MTT and NR tests were more sensitive for the evaluation of BlV cytotoxicity on the VERO E6 cell line.
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暴露于白刺蛇粗毒液的VERO E6细胞的生存能力
原代和连续细胞培养系统是一种很好的体外定量评估蛇毒毒性的方法。目前还很少有研究评价细胞对白须鲸毒(BlV)的反应。本研究旨在评价粗BlV对VERO E6细胞培养的细胞毒性作用。活细胞采用中性红掺入(NR)和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑(MTT)试验,非活细胞采用台盼蓝(TB)染料排斥试验。各组间差异采用单因素方差分析(ANOVA-One-Way), Dunnet通过Graph Pad 5.0程序评估,显著性为5%。BlV 50%细胞毒浓度(CC50)为7.86 μg/mL。在所有评估的测定中,细胞活力都有剂量依赖性降低的趋势。而MTT和NR试验对BlV对VERO E6细胞株的细胞毒性评价更为敏感。
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