CYTO-TOXIC EFFECT OF T-2 MYCOTOXIN ON CELLS IN CULTURE AS DETERMINED BY A RAPID COLORIMETRIC BIOASSAY
CYTO-TOXIC EFFECT OF T-2 MYCOTOXIN ON CELLS IN CULTURE AS DETERMINED BY A RAPID COLORIMETRIC BIOASSAY
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DOI：
10.1016/0041-0101(88)90235-8
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发表时间：
1988-01-01
期刊：
影响因子：
2.8
通讯作者：
DELOACH, JR
中科院分区：
文献类型：
--
作者：
HOLT, PS;BUCKLEY, S;DELOACH, JR
我们开发了一种用于测定暴露于毒性试剂的细胞的代谢活性（活力）的比色测定法。该系统基于活细胞中的线粒体酶将四唑盐修饰成蓝色甲产物的能力，该蓝色甲产物可以在570 nm处通过荧光光度法检测。该测定法对哺乳动物和昆虫细胞系同样有效，在48小时时，颜色形成在1.56-50 × 10 - 6的细胞输入范围内是线性的。104个细胞/ml。T-2真菌毒素对L929细胞中四唑裂解的抑制作用与对蛋白质和DNA合成的抑制作用相当（50%抑制=6-8 ng/ml）。使用该系统分析T-2毒素对来自不同动物物种的细胞的致死作用，发现牛细胞是最敏感的（在2.2ng/ml时50%抑制），而仓鼠细胞是最抗性的（在26.2ng/ml时50%抑制）。小鼠细胞表现出中等敏感性（10.9 ng/ml时抑制率为50%）。在不同的细胞类型中也观察到可变的毒素敏感性。淋巴细胞对T-2抑制作用的敏感性是组织培养细胞系的3倍。这些数据表明，比色测定系统可以在毒理学研究中有广泛的应用。此外，观察到的物种敏感性的差异可能提供洞察T-2毒素-细胞相互作用，最终导致细胞死亡的主要机制。
We developed a colorimetric assay for determining metabolic activity (viability) of cells exposed to toxic agents. This system is based on the ability of mitochondrial enzymes in viable cells to modify a tetrazolium salt into a blue formazan product that can be detected spectrophotometrically at 570 nm. The assay works equally well for mammalian and insect cell lines and at 48 hr color formation is linear over a cell input range of 1.56-50 .times. 104 cells/ml. The inhibitory effects of T-2 mycotoxin on tetrazolium cleavage in L929 cells is comparable to that observed for protein and DNA synthesis (50% inhibition=6-8 ng/ml). Using this system to analyze the lethal effect of T-2 toxin on cells from various animal species, it was found that bovine cells were the most sensitive (50% inhibition at 2.2 ng/ml) while hamster cells were the most resistant (50% inhibition at 26.2 ng/ml). Murine cells exhibited intermediate sensitivity (50% inhibition at 10.9 ng/ml). Variable toxin susceptibility was also observed among different cell types. Lymphocytes were 3-fold more sensitive to the T-2 inhibitory effects than comparable tissue culture cell lines. These data indicate that the colorimetric assay system could have broad applications in toxicological studies. Further, the observed differences in species sensitivity may provide insight into the primary mechanism of the T-2 toxin-cell interaction that ultimately leads to cell death.