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镰刀菌毒素恩镰孢菌素B在体外无遗传毒性活性,但有明显的细胞毒性。

The Fusarium toxin enniatin B exerts no genotoxic activity, but pronounced cytotoxicity in vitro.

作者信息

Behm Claudia, Degen Gisela H, Föllmann Wolfram

机构信息

IfADo, Leibniz-Institut für Arbeitsforschung an der TU Dortmund, Dortmund, Germany.

出版信息

Mol Nutr Food Res. 2009 Apr;53(4):423-30. doi: 10.1002/mnfr.200800183.

Abstract

Enniatin B, a fungal metabolite produced by various Fusarium strains, is a frequent contaminant in cereals used for human foods and animal feeds, but, so far very limited data are available on its toxicity. The aim of this study was to investigate the effects of enniatin B in a battery of short-term tests to evaluate its genotoxic potential. In Salmonella typhimurium assays (Ames assay) with the strains TA 98, TA 100, TA 102, and TA 104, both in the presence and absence of an external metabolizing enzyme system (rat liver S9), no mutagenicity was detected up to toxic levels (100 microM) of enniatin B. Likewise, mutagenicity tests in mammalian cells, i. e., the hypoxanthin-guanin-phosphoribosyl-transferase (HPRT) assay with V79 cells performed with and without S9 mix, did not reveal a significant increase in mutant frequency for enniatin B up to 30 microM, a cytotoxic concentration. Additional tests on other types of genotoxicity, i. e., clastogenicity and chromosomal damage, were conducted in V79 cells, applying the alkaline single cell gel electrophoresis (Comet assay with and without FPG, formamidopyrimidine DNA glycosylase, enzyme) and the micronucleus assay. None of these assays revealed a significant genotoxic potential of enniatin B. However, enniatin B exerts pronounced cytotoxic effects in V79 cells as determined by neutral red uptake assay for 48 h exposure: The IC(20) and IC(50) values of 1.5 and 4 microM, are higher than those of the more potent Fusarium toxin deoxynivalenol (IC(20) 0.6 microM, IC(50) of 0.8 microM), but in a similar range as values reported for cytotoxicity of enniatin B in various tumor cell lines. In summary, despite an apparent lack of genotoxic activity, enniatin B can exert biological activity at low micromolar concentrations in mammalian cells.

摘要

恩镰孢菌素B是由多种镰刀菌菌株产生的一种真菌代谢产物,是人类食品和动物饲料所用谷物中常见的污染物,但迄今为止,关于其毒性的数据非常有限。本研究的目的是通过一系列短期试验研究恩镰孢菌素B的作用,以评估其遗传毒性潜力。在鼠伤寒沙门氏菌试验(艾姆斯试验)中,使用TA 98、TA 100、TA 102和TA 104菌株,无论有无外部代谢酶系统(大鼠肝脏S9),在达到恩镰孢菌素B的毒性水平(100微摩尔)之前均未检测到致突变性。同样,在哺乳动物细胞中的致突变性试验,即在有和没有S9混合物的情况下用V79细胞进行的次黄嘌呤-鸟嘌呤-磷酸核糖转移酶(HPRT)试验,在高达30微摩尔(细胞毒性浓度)的恩镰孢菌素B中,未发现突变频率有显著增加。在V79细胞中进行了关于其他类型遗传毒性的额外试验,即断裂剂性和染色体损伤试验,采用碱性单细胞凝胶电泳(有和没有FPG,甲酰胺嘧啶DNA糖基化酶,酶的彗星试验)和微核试验。这些试验均未显示恩镰孢菌素B有显著的遗传毒性潜力。然而,通过中性红摄取试验测定,恩镰孢菌素B在V79细胞中暴露48小时会产生明显的细胞毒性作用:IC(20)和IC(50)值分别为1.5和4微摩尔,高于毒性更强的镰刀菌毒素脱氧雪腐镰刀菌烯醇(IC(20)0.6微摩尔,IC(50)0.8微摩尔),但与报道的恩镰孢菌素B在各种肿瘤细胞系中的细胞毒性值范围相似。总之,尽管明显缺乏遗传毒性活性,但恩镰孢菌素B在低微摩尔浓度下可在哺乳动物细胞中发挥生物活性。

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