Cytotoxic Effect of Some Mycotoxins and their Combinations on Human Peripheral Blood Mononuclear Cells as Measured by the MTT Assay

S. Stoev, S. Denev, M. Dutton, B. Nkosi
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引用次数: 31

Abstract

The objective of this study was to assess cytotoxic effects of ochratoxin A (OTA), penicillic acid (PA), citrinin (CIT), fumonisin B1 (FB1) and their combinations (as naturally occur in real practice) on human peripheral blood mononuclear cells (PBM) using a simple and cheap "in vitro" cytotoxicity test as MTT assay. A stronger suppression on metabolic activity of phytohaemagglutinin-p (PHA)-stimulated PBM was found for a mixture of the following mycotoxins: OTA, CIT and FB1 as compared to any single mycotoxin. PA was found to increase significantly the metabolic activity of the same cells in concentrations above 111 mg/L. There is no "in vitro" synergistic effect between OTA and PA as measured by MTT assay, unlike the strong "in vivo" synergistic effect between the same mycotoxins. The MTT bioassay is not appropriate to measure cell viability and cell proliferation, especially if metabolic activity of the cells is increased as was observed in PA treated human blood mononuclear cells or if the same is decreased. The doses which exert approximately 50% suppression of metabolic activity of PHA-stimulated human blood mononuclear cells (LD50) are determined to be as follows: for OTA - 1.56 to 3.125 mg/L, CIT - 62.5 to 111 mg/L and FB1 - 62.5 to 500 mg/L. Cell viability (proliferation) of all mycotoxin treated blood mononuclear cells was significantly decreased at the highest concentrations of mycotoxins, but this decrease was significantly stronger for OTA-treated cells and especially for different mixtures of mycotoxins.
MTT法测定某些真菌毒素及其组合对人外周血单个核细胞的细胞毒作用
本研究的目的是评估赭曲霉毒素A (OTA)、青霉酸(PA)、柑橘霉素(CIT)、伏马菌素B1 (FB1)及其组合(在实际应用中自然发生)对人外周血单核细胞(PBM)的细胞毒性作用,采用简单廉价的“体外”细胞毒性试验作为MTT法。与任何单一真菌毒素相比,发现以下真菌毒素混合物对植物血凝素-p (PHA)刺激的PBM代谢活性的抑制更强:OTA, CIT和FB1。当浓度超过111 mg/L时,发现PA显著增加了相同细胞的代谢活性。通过MTT测定,OTA和PA之间不存在“体外”协同效应,而同一种真菌毒素之间存在强烈的“体内”协同效应。MTT生物测定法不适用于测量细胞活力和细胞增殖,特别是如果细胞的代谢活性增加,如在PA处理的人血液单核细胞中观察到的那样,或者如果代谢活性降低。对pha刺激的人血单核细胞(LD50)的代谢活性产生约50%抑制的剂量确定如下:OTA - 1.56至3.125 mg/L, CIT - 62.5至111 mg/L, FB1 - 62.5至500 mg/L。在霉菌毒素浓度最高的情况下,所有霉菌毒素处理过的血液单核细胞的细胞活力(增殖)都显著降低,但ota处理过的细胞,尤其是不同真菌毒素混合物处理过的细胞,这种降低明显更强。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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