IRTA, Ctra. Poble Nou, km 5.5, 43540 Sant Carles de la Ràpita, Tarragona, Spain.
Toxicon. 2010 Feb-Mar;55(2-3):381-9. doi: 10.1016/j.toxicon.2009.08.017. Epub 2009 Sep 3.
New advances in the use of NG108-15 cells for marine toxins detection and quantification are presented. We have established modifications on ouabain and veratridine proportion to obtain good toxins dose-response curves on this cell line for neurotoxins acting on voltage gated sodium channel (VGSC). Interesting, differences in the toxic response were observed between two VGSC activating toxins, brevetoxin-3 and pacific ciguatoxin-1. For non-VGSC acting lipophilic toxins, several factors that may influence toxin detection and quantification were analyzed. One hour cultures and forty-eight hours of exposure time, compared with 24h of culture and 24h of exposure, would increase NG108-15 cell maximal yessotoxin (YTX) and azaspiracid-1 (AZA-1) toxic response whereas no change was observed for okadaic acid (OA), dinophysistoxin-1 and pectenotoxin-2. Dose-response curves obtained for YTX or AZA-1 showed variability according to the day of the experiment while good reproducibility was obtained for OA. Evaporation time of toxin solutions before cell exposure could be an important source of variability in AZA-1 toxic response evaluation.
本文介绍了 NG108-15 细胞在海洋毒素检测和定量中的新进展。我们对哇巴因和藜芦碱进行了修饰,以在这条细胞系上获得良好的毒素剂量反应曲线,用于作用于电压门控钠离子通道(VGSC)的神经毒素。有趣的是,两种激活 VGSC 的毒素——布雷毒素-3 和太平洋雪卡毒素-1 的毒性反应存在差异。对于非 VGSC 作用的亲脂性毒素,分析了可能影响毒素检测和定量的几个因素。与 24 小时培养和 24 小时暴露相比,1 小时培养和 48 小时暴露会增加 NG108-15 细胞对 yessotoxin(YTX)和azaspiracid-1(AZA-1)的最大毒性反应,而对 okadaic acid(OA)、dinophysistoxin-1 和pectenotoxin-2 则没有变化。YTX 或 AZA-1 的剂量反应曲线根据实验日期而有所不同,而 OA 的重现性良好。毒素溶液在细胞暴露前的蒸发时间可能是 AZA-1 毒性反应评估中变异性的一个重要来源。