黄曲霉毒素B_(1)、玉米赤霉烯酮和呕吐毒素的累加细胞毒性研究

Study on accumulative cytotoxicity of aflatoxin B_(1),zearalenone and deoxynivalenol

由于饲料中多种霉菌毒素并存的几率比较高,本研究以仔猪肠上皮细胞(IPEC-J2)为模型,研究黄曲霉毒素B1(AFB_(1))、玉米赤霉烯酮(ZEA)和呕吐毒素(DON)的叠加细胞毒性。细胞毒性试验选用AFB1、ZEA和DON三种毒素作为响应面Box-Behnke设计的三个因素,以AFB_(1):10、20、30μg/L,ZEA:150、300、450μg/L,DON:500、1000、1500μg/L作为Box-Behnke设计的三个编码水平。利用响应面设计构建得到17组复合霉菌毒素组合,以其对IPEC-J2细胞活力的影响作为参考指标,得到对细胞损伤程度最高和最低的霉菌毒素添加比例。结果表明:经方程预测后,得到细胞活力最低(霉菌毒素毒性最高)的AFB_(1)、ZEA和DON组合为30、150μg/L和1500μg/L,经测定细胞活力为32.32%;得到细胞活力最高(霉菌毒素毒性最低)的AFB_(1)、ZEA和DON组合为10、150μg/L和600μg/L,经测定细胞活力为53.01%。该结果为多种霉菌毒素叠加毒性的研究提供了依据。

Because many kinds of mycotoxins coexist in feedstuffs or animal diets,swine jejunal epithelial cells(IPEC-J2)were used to study the accumulative cytotoxicity of aflatoxin B1(AFB1),zearalenone(ZEA)and deoxynivalenol(DON).AFB_(1),ZEA and DON were selected as the three factors of Box-Behnke design.The different mycotoxin concentrations of 10,20,30μg/L for AFB_(1),150,300,450μg/L for ZEA,500,1000,1500μg/L for DON were used as the three coding levels of Box-Behnke design.17 groups of combined mycotoxin combinations were constructed by response surface design,and their effects on IPEC-J2 cell viability were taken as the reference indicators to obtain the highest and lowest cell viability caused by the different mycotoxin combinations.The results showed that the lowest cell viability(the highest cytotoxicity)was 32.32%when AFB_(1),ZEA and DON concentrations were 30,150 and 1500μg/L,while the highest cell viability(the lowest cytotoxicity)was 53.01%when AFB1,ZEA and DON concentrations were 10,150 and 600μg/L.This result provided an effective strategy to study the accumulative cytotoxicity of mycotoxins.