Department of Pharmacology & Toxicology, National Veterinary Research Institute, Al. Partyzantow 57, 24-100 Pulawy, Poland.
Int J Mol Sci. 2009 Jan;10(1):37-49. doi: 10.3390/ijms10010037. Epub 2008 Dec 27.
Embryonic midbrain micromass cultures were exposed for five days to ochratoxin A (OTA) at seven concentrations (ranging from 0.16 to 10 microg/mL). Cell viability was assessed in neutral red uptake test (NRU), and differentiation - by immunoenzymatic determination of structural proteins (beta(III)-tubulin, MAP2, GFAP) expression level as well as by computer image analysis. Dose dependent decrease in cell number and differentiation was observed. Concentration-response curves were analysed and the mean inhibition concentrations (microg/mL) for cytotoxicity (IC(50)) and differentiation (ID(50)) were calculated. There were no significant differences in the sensitivity of neurons in early and late stage of differentiation and astrocytes to the toxic activity of this compound. For all endpoints ID(50) value was very low (< 10 microg/mL) so OTA was classified as a strong teratogen. IC(50)/ ID(50) ratios <2 pointed out that with harmful action of OTA the basic cytotoxicity should be connected.
胚胎中脑组织微团培养物在五种浓度的黄曲霉毒素 A(OTA)下暴露 5 天(范围从 0.16 到 10 微克/毫升)。通过中性红摄取试验(NRU)评估细胞活力,通过免疫酶确定结构蛋白(β(III)-微管蛋白、MAP2、GFAP)的表达水平,以及通过计算机图像分析来评估分化。观察到细胞数量和分化程度的剂量依赖性下降。分析浓度反应曲线,并计算细胞毒性(IC(50))和分化(ID(50))的平均抑制浓度(微克/毫升)。在对这种化合物的毒性活性方面,早期和晚期分化的神经元和星形胶质细胞的敏感性没有显著差异。对于所有终点,ID(50)值非常低(<10 微克/毫升),因此 OTA 被归类为强致畸物。IC(50)/ID(50)比值<2 表明 OTA 的有害作用应与基本细胞毒性有关。