First-line Anti-tuberculosis Drug-induced Liver Injury In Mice On The Nf-κb Expression In Liver Cells

Objectives To observe the expressions of Nuclear factor kappaB(NF-κB), Nuclear factor kappa B inhibitor(IκB) and Tumor necrosis factor alpha(TNF-α) which induces by anti-tuberculosis drugs in mice liver. To reveal the changes of NF-κB in the development and progression of liver injury.Methods A total of 168 clean Kunming mice were randomly divided into 4experimental groups and one baseline group. The 4 experimental groups were given isoniazid(90mg/Kg.d), rifampicin(135mg/Kg.d), pyrazinamide(315mg/Kg.d) and three medicine combined(90mg/Kg.d+135mg/Kg.d+315mg/Kg.d) orally at 3d, 5d, 7d, 10 d,15d and the baseline group was put to death before started experimental one day. Serum ALT, AST levels were determined using an automatic biochemical analyzer, and MDA contents and SOD activities were detected by biochemical method. Enzyme-Linked immunosorbant assay was used to test the protein contents of NF-κB, IκB and TNF-α.SYBR Green Real-time RT-PCR was used to test the expressions of NF-κB, IκB and TNF-α m RNA. Distributed measurement data was discribed by mean±standard deviation( x ±s). Factor analysis was used to test the effect of interaction between group and time. the comparison of various of mean all used Brown-Forsythe, the comparison between two groups used Dunnett `s T3 methods. Correlation analysis used Spearman correlation analysis, in which, if P<0.05, the disparity had statistical significance.Results The results of hepatic histopathological changes and the general liver function index for clinical diagnosis of liver disease showed: Hepatocytes changed after 3d and the ALT and AST levels of treated groups was increased significantly, suggested that successful model. Compared with the baseline group, MDA content of liver tissue in mice treated groups gradually increased, while SOD activity was decreased, they were negatively correlated. Further the inhibition of NF-κB activity and m RNA, protein expression of inflammation-related factors factor IκB and TNF-α were analysised. In the discovery, with the change of NF-κB, IκB and TNF-α m RNA expression, the corresponding protein expression also showed a trend of increase accordingly. Factor analysis was used to test the m RNA and protein expression of NF-κB and TNF-α, the comparisons of four groups had statistical significance(NF-κB: F=3.110, P=0.028;F=18.551, P<0.001. TNF-α: F=8.655, P<0.001; F=8.218, P=0.004), the comparisons at the different time points had statistical significance(NF-κB: F=4.944, P<0.001; F=5.920,P<0.001. TNF-α: F=6.171, P<0.001; F=11.807, P<0.001)and the effect of interaction between group and time was statistically significant(NF-κB: F=1.744, P=0.047; F=4.596,P<0.001. TNF-α: F=2.756, P=0.001; F=4.495, P<0.001). With prolonged treatment, the trend of the m RNA and protein expression of NF-κB and TNF-α was consistent in each group, isoniazid group gradually increased while rifampin group, pyrazinamide group and the three-drug combination group NF-κB m RNA and protein appeared first increased and then decreased, however, the opposite trend in the m RNA and protein expression of IκB in each group, isoniazid group showed a trend of decreased as a whole, rifampin group, pyrazinamide group and three drug combination group increased after the first decline trend.Conclusions In the development of liver injury by Isoniazid, the NF-κB pathway is activated, IκB gene expression down-regulated and TNF-αgene expression up-regulated;rifampicin and pyrazinamide itself has immunomodulatory effect that makes the NF-κB pathway activation is restrained, NF-κB activation of three medicine combined group may be a combination of the promotional effects of isoniazid and inhibitory effect of rifampicin and pyrazinamide.