Abal Paula, Louzao M Carmen, Fraga María, Vilariño Natalia, Ferreiro Sara, Vieytes Mercedes R, Botana Luis M
Departamento de Farmacología, Facultad de Veterinaria, Universidad de Santiago de Compostela, Lugo, Spain.
Departamento de Fisiología, Facultad de Veterinaria, Universidad de Santiago de Compostela, Lugo, Spain.
Cell Physiol Biochem. 2017;43(1):136-146. doi: 10.1159/000480331. Epub 2017 Aug 28.
Azaspiracids (AZAs) are marine biotoxins produced by the dinoflagellates genera Azadinium and Amphidoma. These toxins cause azaspiracid poisoning (AZP), characterized by severe gastrointestinal illness in humans after the consumption of bivalve molluscs contaminated with AZAs. The main aim of the present study was to examine the consequences of human exposure to AZA1 by the study of absorption and effects of the toxin on Caco-2 cells, a reliable model of the human intestine.
The ability of AZA1 to cross the human intestinal epithelium has been evaluated by the Caco-2 transepithelial permeability assay. The toxin has been detected and quantified using a microsphere-based immunoassay. Cell alterations and ultrastructural effects has been observed with confocal and transmission electron microscopy Results: AZA1 was absorbed by Caco-2 cells in a dose-dependent way without affecting cell viability. However, modifications on occludin distribution detected by confocal microscopy imaging indicated a possible monolayer integrity disruption. Nevertheless, transmission electron microscopy imaging revealed ultrastructural damages at the nucleus and mitochondria with autophagosomes in the cytoplasm, however, tight junctions and microvilli remained unaffected.
After the ingestion of molluscs with the AZA1, the toxin will be transported through the human intestinal barrier to blood causing damage on epithelial cells.
azaspiracids(AZAs)是由裸甲藻属和尖尾藻属的甲藻产生的海洋生物毒素。这些毒素会导致azaspiracid中毒(AZP),其特征是人类食用受AZAs污染的双壳贝类后出现严重的胃肠道疾病。本研究的主要目的是通过研究毒素对人肠道的可靠模型Caco-2细胞的吸收和影响,来检验人体接触AZA1的后果。
通过Caco-2跨上皮通透性试验评估AZA1穿过人肠上皮的能力。使用基于微球的免疫测定法检测和定量毒素。用共聚焦显微镜和透射电子显微镜观察细胞变化和超微结构效应。结果:AZA1以剂量依赖性方式被Caco-2细胞吸收,且不影响细胞活力。然而,共聚焦显微镜成像检测到的闭合蛋白分布变化表明可能存在单层完整性破坏。尽管如此,透射电子显微镜成像显示细胞核和线粒体有超微结构损伤,细胞质中有自噬体,然而,紧密连接和微绒毛未受影响。
摄入含有AZA1的软体动物后,毒素将通过人体肠道屏障运输到血液中,对上皮细胞造成损伤。