黄曲霉毒素B_1致体外人胚肝细胞DNA损伤的初步研究

Preliminary study on the DNA damage of human embryo liver cell in vitro induced by aflatoxin B_1

目的研究黄曲霉毒素B1(AFB1)对人胚肝细胞(L-02细胞)的毒性作用特点,为建立AFB1致L-02细胞DNA损伤的体外实验模型打下基础。方法以L-02细胞为靶细胞,采用完全随机实验设计将其分为3组:(1)空白对照组;(2)溶剂对照组;(3)AFB1染毒组,分别使用5,10,20,40mg/L 4个剂量染毒。各组细胞染毒培养24h后,收集细胞进行单细胞凝胶电泳(SCGE)试验,观察细胞DNA损伤情况;同时收集细胞培养上清液,用全自动生化分析仪测定谷草转氨酶(AST)和谷丙转氨酶(ALT)含量。结果(1)SCGE试验结果显示,染毒浓度达到40mg/L时L-02细胞出现DNA损伤,其尾长和Olive尾矩值与空白对照和溶剂对照相比有显著差异(P<0.01,P<0.05);(2)各AFB1染毒组细胞培养上清液的AST和ALT含量,分别与空白对照组比较均未出现显著性差异(P>0.05)。结论 AFB1染毒浓度达到40mg/L的浓度时,就能够诱导L-02细胞DNA出现损伤,但肝功能AST和ALT却未见异常。实验结果表明,在L-02细胞未出现急性损伤的低浓度AFB1染毒状态下,其致肝细胞DNA损伤的作用就已显现。

Objective To study the toxic effect of aflatoxin B1 on the human embryo liver cell（L-02 cell） in order to set up an in vitro model of L-02 cell DNA damage induced by AFB 1.Methods L-02 cell was choosed as a target cell,and using a completely randomized design,the cell was divided into three groups：（Ⅰ）blank control group;（Ⅱ）solvent control group;（Ⅲ）AFB 1 exposed groups,with 5,10,20,40mg/L four dosage contaminations.After each group of cells was cultured for 24 hours,SCGE was performed on the collected cell,and the situation of cell damage was also observed.At the same time,the cell culture supernatant of each group was collected,the values of AST and ALT were determined by the automatic biochemistry analyzer.Results（Ⅰ） SCGE showed that only 40mg/L exposure group occurred a cell DNA damage,Taillength and Olive tailmoment in this group（P〈0.01,P〈0.05） compared with blank control and solvent control groups were significantly different.（Ⅱ） The values of AST and ALT in cell culture supernatant of each exposed group were not significant differences（P〉0.05） compared with blank control group.Conclusion SCGE has shown that 40mg/L AFB 1 is able to induce cell DNA damage;but the values of AST and ALT are not obvious changes.Experimental results indicate that cell DNA damage has occurred even then acute cell injury induced by low concentration of AFB 1.has not been observed.