黄曲霉毒素B1与杂色曲霉素对HepG2细胞的联合毒性

Multi-index Detection of Toxicoligical Interaction Between the Aflatoxin B1 and Sterigmatocystin in HepG2 Cells

摘要:

研究了黄曲霉毒素B1 (AFB1)和杂色曲霉素(ST)对人肝癌细胞HepG2的细胞毒性及其联合毒性。体外培养人肝癌细胞HepG2, 用不同浓度梯度的AFB1 (0.1、1、5、10 μmol/L)和ST(0.5、2.5、5、7 μmol/L)单独和联合作用于HepG2细胞24 h或48 h, 分别测量细胞活性、ATP含量、DNA受损程度、线粒体膜电位, 以及活性氧含量的变化, 通过统计分析得到两种毒素对HepG2细胞的联合毒性作用。结果显示, AFB1和ST对HepG2细胞的IC50分别为16.989 μmol/L和7.258 μmol/L, 细胞增殖力、ATP、DNA、线粒体膜电位、活性氧等指标的理论加和值与实际值无显著性差异(P>0.05)。同时, 这两种毒素及其混合物对HepG2细胞的作用终点均可分为3类：第一类为细胞增殖力的降低; 第二类包括ATP含量、DNA含量变化; 第三类为活性氧含量变化和线粒体膜通透性变化。表明AFB1和ST对HepG2细胞的联合毒性作用为加和作用。

Abstract:

Aflatoxin B1 (AFB1)and Sterigmatocystin(ST) are two fungal toxins that are commonly coexisted in many crops, and their joint cytotoxicity to human hepatoma cell line HepG2 was investigated. HepG2 was treated with different concentration of AFB1(0.1, 1, 5, 10 μmol/L), ST(0.5, 2.5, 5, 7 μmol/L) and their mixture for 24 h or 48 h to analyze their combinative toxic effect through measuring the toxicity end points including change of cell proliferation, ATP content, degree of DNA damage, change of mitochondrial membrane potential and active oxygen content. The IC50 of AFB1 and ST was found to be 16.989 μmol/L and 7.258 μmol/L, respectively. There was no significant difference(P>0.05)between the theoretical value compared with the actual value of all the end points.In the mean time, the end points of cytotoxicity can be divided into three categories by principle component analysis(PCA)：the first category is the cell proliferation, the second class includes ATP content, DNA content; the third class is the active oxygen content and the mitochondrial permeability transition. The results indicate that interaction type between AFB1 and ST is additive joint effect.