| Objective:This study through of Procyanidin B2(PC-B2) resist Aflatoxin B1(AFB1) on human embryonic stem cells (L-02) hepatocyte oxidative damage and influence of the phase I metabolizing enzymes CYP1A2,3A4 activity and the gene expression, to explore the mechanism and effect of liver cell injury induced by AFB1 intervention PC-B2, initially clear PC-B2 on liver cell protective effect, for the future further protect liver cancer treatment and prevention of provide the scientific basis.Methods:Using in vitro cell experiment, embryo liver cells (L-02 cells) in logarithmic phase are treat with PC-B2 and/or AFB1. The experiment was divided into five groups. Set the concentration 30μg/ml for AFB1 infection; 3,10, 30μg/ml for PC-B2 experimental treatment levels.MTT method is used to test the cell vitality, and observe cell morphology by inverted fluorescence microscope.Using kits test the levels of MDA, SOD, gsh-px, and the enzyme activity of CYP1A2 and CYP3A4. Gene mRNA expression level was test through the method of qPCR. Using SPSS 16.0 software for analysis.Results:Compared with solvent control group, group treat with 30μg/ml AFB1, floating cells increased, the cell vitality (%) significantly suppressed (69.9 ± 2.46) (P<0.05);Group treat with AFB1 and 30μg/ml PC-B2,cell activity significantly increased (P<0.05).Compared with solvent control group, group treat with 30μg/ml AFB1, MDA generation increased significantly (P<0.05), but after treat with different concentration of PC-B2, MDA generation decreased (P<0.05).Group treat with AFB1, cells’SOD and gsh-px activities (U/mgprot) were 35.96±0.83 and 35.96±9.98, compared with solvent control group 40.01±0.54, 287.10±6.96 were significantly decreased (P<0.05);Group treat with both AFB1 and 30μg/ml PC-B2 can significantly inhibit the trend.Cells’enzyme activity of CYP1A2 and CYP3A4 were increased in group treat with 30μg/ml AFB1(P<0.05);And PC-B2 intervention group L-02 cells’s CYP1A2 and CYP3A4 enzyme activity compared with AFB1 infected group reduced significantly (P<0.05), and high-dose PC-B2 intervention’s influence on the enzyme activity more obvious (P<0.05).Compared with solvent control group,the CYP1A2, CYP3A4 gene expression of L-02 cells with AFB1 for 24h increased(P<0.05).PC-B2 intervention could inhibit the gene expression significantly (P<0.05), and were dependent on the does within a certain concentration range.Conclusion:1. AFB1 inhibited the growth of liver cell, and PC-B2 can promote the cell growth in a certain concentration range, and can reduce the AFB1 exposure caused by the cell survival rate decreased.2. PC-B2 reduced AFB1 induced damage to the liver cell antioxidant system, inhibited AFB1 induced cell MDA content increased and SOD, GSH-PX enzyme activity decreased.3. After AFB1 inhibition, the cell’s CYP1A2 and CYP3A4 enzyme activity increased, PC-B2 intervention can inhibit the increase of these two enzymes activity, suggesting that AFB1 may play a role in the metabolism of the body through the intervention of PC-B2.4. PC-B2 has the role of anti AFB1 induced liver cell damage, it is possible to become the selection of chemical prevention in high pollution area of aflatoxin. |
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