Department of Life Science, University of Trieste , 34127 Trieste, Italy.
Department of Pharmacy, University of Napoli Federico II , 80131 Napoli, Italy.
Environ Sci Technol. 2016 Feb 2;50(3):1544-51. doi: 10.1021/acs.est.5b04749. Epub 2016 Jan 12.
This study provides the first evaluation of the cytotoxic effects of the recently identified palytoxin (PLTX) analog, ovatoxin-a (OVTX-a), the major toxin produced by Ostreopsis cf. ovata in the Mediterranean Sea. Its increasing detection during Ostreopsis blooms and in seafood highlights the need to characterize its toxic effects and to set up appropriate detection methods. OVTX-a is about 100 fold less potent than PLTX in reducing HaCaT cells viability (EC50 = 1.1 × 10(-9) M vs 1.8 × 10(-11) M, MTT test) in agreement with a reduced binding affinity (Kd = 1.2 × 10(-9) vs 2.7 × 10(-11) M, saturation experiments on intact cells). Similarly, OVTX-a hemolytic effect is lower than that of the reference PLTX compound. Ost-D shows the lowest cytotoxicity toward HaCaT keratinocytes, suggesting the lack of a hydroxyl group at C44 as a critical feature for PLTXs cytotoxic effects. A sandwich ELISA developed for PLTX detects also OVTX-a in a sensitive (LOD = 4.2 and LOQ = 5.6 ng/mL) and accurate manner (Bias = 0.3%), also in O. cf. ovata extracts and contaminated mussels. Although in vitro OVTX-a appears less toxic than PLTX, its cytotoxicity at nanomolar concentrations after short exposure time rises some concern for human health. The sandwich ELISA can be a viable screening method for OVTXs detection in monitoring program.
本研究首次评估了最近发现的石房蛤毒素(PLTX)类似物,卵形毒素-a(OVTX-a)的细胞毒性作用,OVTX-a 是地中海拟菱形藻中产的主要毒素。由于在拟菱形藻水华期间和海产品中检测到其含量不断增加,因此需要对其毒性作用进行特征描述,并建立适当的检测方法。OVTX-a 在降低 HaCaT 细胞活力方面的效力比 PLTX 低约 100 倍(EC50 = 1.1×10(-9) M 对 1.8×10(-11) M,MTT 试验),与结合亲和力降低(Kd = 1.2×10(-9) 对 2.7×10(-11) M,完整细胞的饱和实验)一致。同样,OVTX-a 的溶血作用低于参考 PLTX 化合物。Ost-D 对 HaCaT 角质形成细胞的细胞毒性最低,表明 C44 上缺少羟基是 PLTX 细胞毒性作用的关键特征。为 PLTX 开发的夹心 ELISA 以敏感(LOD = 4.2 和 LOQ = 5.6 ng/mL)和准确的方式(偏差 = 0.3%)检测 OVTX-a,也可检测到拟菱形藻提取物和受污染贻贝中的 OVTX-a。尽管 OVTX-a 在体外的毒性似乎低于 PLTX,但在短时间暴露于纳摩尔浓度下,其细胞毒性引起了人们对人类健康的关注。夹心 ELISA 可以作为监测计划中检测 OVTXs 的可行筛选方法。