Clarke R, Connolly L, Frizzell C, Elliott C T
Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Northern Ireland, United Kingdom.
Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Northern Ireland, United Kingdom.
Toxicol Lett. 2015 Mar 18;233(3):278-86. doi: 10.1016/j.toxlet.2015.01.011. Epub 2015 Jan 23.
Aflatoxin B1 (AFB1), ochratoxin A (OTA) and fumonisin B1 (FB1) are important mycotoxins in terms of human exposure via food, their toxicity and regulatory limits that exist worldwide. Mixtures of toxins can frequently be present in foods, however due to the complications of determining their combined toxicity, legal limits of exposure are determined for single compounds, based on long standing toxicological techniques. High content analysis (HCA) may be a useful tool to determine total toxicity of complex mixtures of mycotoxins. Endpoints including cell number (CN), nuclear intensity (NI), nuclear area (NA), plasma membrane permeability (PMP), mitochondrial membrane potential (MMP) and mitochondrial mass (MM) were compared to the conventional 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and neutral red (NR) endpoints in MDBK cells. Individual concentrations of each mycotoxin (OTA 3 μg/ml, FB1 8 μg/ml and AFB1 1.28 μg/ml) revealed no cytotoxicity with MTT or NR but HCA showed significant cytotoxic effects up to 41.6% (p≤0.001) and 10.1% (p≤0.05) for OTA and AFB1, respectively. The tertiary mixture (OTA 3 μg/ml, FB1 8 μg/ml and AFB1 1.28 μg/ml) detected up to 37.3% and 49.8% more cytotoxicity using HCA over MTT and NR, respectively. Whilst binary combinations of OTA (3 μg/ml) and FB1 (8 μg/ml) revealed synergistic interactions using HCA (MMP, MM, NI endpoints) not detected using MTT or NR. HCA is a highly novel and sensitive tool that could substantially help determine future regulatory limits, for single and combined toxins present in food, ensuring legislation is based on true risks to human health exposure.
就人类通过食物接触黄曲霉毒素B1（AFB1）、赭曲霉毒素A（OTA）和伏马菌素B1（FB1）而言，这些都是重要的霉菌毒素，它们具有毒性，且在全球范围内都设有监管限值。食物中常常会出现毒素混合物，但由于确定其联合毒性存在复杂性，基于长期的毒理学技术，针对单一化合物确定了法定接触限值。高内涵分析（HCA）可能是一种用于确定霉菌毒素复杂混合物总毒性的有用工具。将包括细胞数量（CN）、核强度（NI）、核面积（NA）、质膜通透性（PMP）、线粒体膜电位（MMP）和线粒体质量（MM）在内的终点指标，与MDBK细胞中传统的3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）和中性红（NR）终点指标进行了比较。每种霉菌毒素的单独浓度（OTA 3μg/ml、FB1 8μg/ml和AFB1 1.28μg/ml）用MTT或NR检测时均未显示出细胞毒性，但HCA显示OTA和AFB1分别有高达41.6%（p≤0.001）和10.1%（p≤0.05）的显著细胞毒性作用。三元混合物（OTA 3μg/ml、FB1 8μg/ml和AFB1 1.28μg/ml）用HCA检测时，分别比MTT和NR检测到的细胞毒性高出37.3%和49.8%。虽然OTA（3μg/ml）和FB1（8μg/ml）的二元组合用HCA（MMP、MM、NI终点指标）检测时显示出协同相互作用，但用MTT或NR未检测到。HCA是一种非常新颖且灵敏的工具，可极大地有助于确定未来食物中单一和混合毒素的监管限值，确保立法基于对人类健康接触的真实风险。
