Preliminary In Vitro Study On The Damage Of Human Embryo Liver Cell Induced By Aflatoxin B1

Aflatoxins is one kind has the greatly strengthened toxic Secondary Metabolites of fungus, has created extremely serious threat to the human health. In natural pollution food by Aflatoxin B1(AFB1) most sees, its toxicity and carcinogenicity is also strongest. When the human takes in massively, has the possibility to be able to have the acute poison, has the acute hepatitis, the hemorrhagic necrosis, the liver cell fat denaturation and the bile duct proliferation situation, etc. When micro continues to take in, may create the chronic poison, the growth barrier, causes the thready pathological change and the fiber structure proliferation, etc. In view of above characteristic, for further explores AFB1and between the liver damage relations, this topic has conducted following research.Objective Discuss toxic effect to the Human embryo liver cell(L-02cell) induced by Aflatoxins B1, set up an in vitro model of L-02cell DNA injury, provide reference for study the preventive intervention of AFB1.Methods Take the L-02cell as the target cell, uses the completely randomized design in vitro experiment. Carries on the routine culture to L-02cell, the selection is in the logarithmic growth phase and grows the good cell to carry on the experiment.(â… ) MTT assay:The experiment sets up the blank control group, the solvent control group, the AFB1contamination group, with5,10,20,40,80,160,320mg/L seven dosage contaminations. Cultures after for24hours in each group of cells, carries on the cell MTT assay, observe the cell activity and apoptosis of each group.(II) Single cell gelelectrophoresis assay(SCGE):Grouping random experiment, sets up the blank and the solvent control group, the AFB1contamination group, with5,10,20,40mg/L four dosage contaminations. Cultures after for24hours in each group of cells, carries on SCGE assay, observe the situation of cell damage. At the same time, simultaneously collects the cell culture supernatant of each group, determines by the automatic biochemistry analyzer to analy aspartate aminotransferase (AST) and alanine aminotransferase (ALT).Results (I) MTT assay showed that each exposure group compared with blank control significant difference (P<0.01), cell toxicity was dose-response relationship.(II)SCGE showed that only40mg/L exposure groups showing a cell damage, Taillength (P<0.01) and Olivetailmoment (P<0.05) compared with blank control were significantly different.(â…¢) The cell culture supernatant of each exposure group of AST and ALT there were no significant differences (P>0.05) compared with blank control.Conclusion MTT assay of exposure doses above80mg/L or more, can present significant cytotoxic effect, cause the cell viability decreased significantly. SCGE have shown that40mg/L AFB1was able to induce the L-02cell DNA damage; but the value of AST and ALT had no abnormal finding. Test results indicate that DNA damage occurred when had not be found acute injury of cell induced by low concentration of AFB1. Intake less AFB1during prolonged would have induced cancer. AFB1induced acute injury of cultured cells in vitro at home and abroad are few reports of acute toxic effects have yet to be further explored, so chose L-02cell and AFB1in this research. The acute toxic effect of AFB1that needs to be further investigated research, in order to set up in vitro cell model.