Hanelt M, Gareis M, Kollarczik B
Institute of Medical Microbiology, Infectious and Epidemic Diseases, Veterinary Faculty, Ludwig-Maximilian-University Munich, Germany.
Mycopathologia. 1994 Dec;128(3):167-74. doi: 10.1007/BF01138479.
The application of a modified colorimetric bioassay for the evaluation of the biological effects of mycotoxins is reported. Using three different monolayer cell lines (swine kidney, Madin Darby canine kidney, HeLa) the influence of nine different mycotoxins on the cellular methylthiazoltetrazolium (MTT)-cleavage activity was evaluated. The yellow tetrazolium salt MTT is converted by mitochondrial dehydrogenases of metabolically active cells to an insoluble purple formazan product, which was then solubilized with dimethylsulfoxide. The optical density of this homogeneous solution was suitable for a precise spectrophotometric measurement by a plate reader at a wavelength of 510 nm. Nine mycotoxins were simultaneously tested in all three cell lines, from which the swine kidney cell line proved to be the most sensitive. The effects of additional 35 mycotoxins were therefore tested using swine kidney monolayers as target cells. A total of 28 toxins of the 44 mycotoxins tested proved to be cytotoxic in the MTT-bioassay. Most of them belong to the group of trichothecene mycotoxins. Concentrations ranged between 0.01 micrograms and 100 micrograms/ml of cell culture medium. The MTT cleavage assay was found to be a quick (24 hours) and easy to perform system for the evaluation of the biological activity of many different mycotoxins and may also provide a useful tool for the testing of a large variety of sample materials.
本文报道了一种改良的比色生物测定法在评估霉菌毒素生物学效应中的应用。使用三种不同的单层细胞系(猪肾细胞、马迪达比犬肾细胞、人宫颈癌HeLa细胞),评估了九种不同霉菌毒素对细胞甲基噻唑基四氮唑(MTT)裂解活性的影响。黄色的四氮唑盐MTT被代谢活跃细胞的线粒体脱氢酶转化为不溶性的紫色甲臜产物,然后用二甲基亚砜溶解。这种均匀溶液的光密度适合用酶标仪在510nm波长下进行精确的分光光度测量。在所有三种细胞系中同时测试了九种霉菌毒素,结果证明猪肾细胞系最为敏感。因此,使用猪肾单层细胞作为靶细胞,测试了另外35种霉菌毒素的作用。在所测试的44种霉菌毒素中,共有28种毒素在MTT生物测定中被证明具有细胞毒性。它们中的大多数属于单端孢霉烯族霉菌毒素。浓度范围在0.01微克至100微克/毫升细胞培养基之间。发现MTT裂解测定法是一种快速(24小时)且易于操作的系统,用于评估许多不同霉菌毒素的生物活性,也可能为测试多种样品材料提供有用的工具。