Study Of HMGB1-RAGE Signal Pathway In The Prevention Of Anti-Tuberculosis Drug-Induced Liver Injury With Orazamide

Objective:To explore the role and possible mechanism of the HMGB1-RAGE signaling pathway in the prevention of anti-tuberculosis drug-induced liver injury with Orazamide.Methods:Sixty male Kunming mice were equally and randomly divided into 6 groups:a blank group,a model group,a low-dose of orazamide group（ALMD-L group）,a medium-dose of Orazamide group（ALMD-M group）,a high-dose of orazamide group（ALMD-H group）and a positive control group,each with 10 mice.Except for the blank group,each group was given isoniazid 75mg·kg^-1·d^-1+rifampicin75mg·kg^-1·d^-1.In addition to the model group,each treatment group was given corresponding drug intervention:the positive control group was given diammonium glycyrrhizinate 60mg·kg^-1·d^-1,ALMD-L group were given orazamide 80mg·kg^-1·d^-1,ALMD-M group 160mg·kg^-1·d^-1,ALMD-H group 320mg·kg^-1·d^-1 separately.After 14 days of continuous administration,blood was taken from eyeballs of mice in each group to measure serum liver function by using automatic biochemical analyzer,including Alanine Aminotransferase、Aspartate Aminotransferase、Total Bilirubin、Direct Bilirubin、Alkaline Phosphatase、γ-Glutamyl Transpeptidase and Total Bile Acids.Livers were taken to calculate liver index,and to observe pathological changes by HE staining.Expression of High Mobility Group Box-1 and Nuclear FactorκB in liver tissue were detected by immunohistochemistry.Expression of Receptor for Advanced Glycation End Products and Tumor Necrosisi Factor alpha were detected by enzyme-linked immunosorbent assay.Moreover,The correlation analysis between the protein expression levels of HMGB1 and RAGE in liver tissues and the levels of ALT and TBIL in serum of mice in the model group were carried out.Results:1.Liver index results:Compared with the blank group,the liver index of mice in the model group increased（P<0.05）;compared with the model group,the liver index of mice in each treatment group decreased（P<0.05）.However,there was no statistically significant difference between the treatment groups（P>0.05）.2.Liver function results:Compared with the blank group,the levels of TBIL,DBIL,ALT,AST,AKP,TBA andγ-GT in serum of the model group increased（P<0.05）;compared with the model group,the liver function of each treatment group improved（P<0.05）.3.HE staining results:Compared with the blank group,the mice in the model group had disordered liver cord arrangement,degeneration and necrosis of liver cell,which was most obvious adjacent to the central vein and accompanied by obvious inflammatory cell infiltration;compared with the model group,the liver lobule structure and inflammatory cell infiltration in each treatment group showed different degrees of improvement.The liver tissue structure of the ALMD-H group and the positive control group was the closest to the blank group.4.Immunohistochemical results:Compared with the blank group,the protein expression of HMGB1 and NF-κB in the liver tissue of mice in the model group increased（P<0.05）;compared with the model group,the protein expression of HMGB1 and NF-κB in the liver tissue of mice in each treatment group decreased（P<0.05）.Compared with ALMD-L and ALMD-M groups,the ALMD-H group had the most significant decrease（P<0.05）,and the difference was not statistically significant compared with the positive control group（P>0.05）.5.ELISA results:Compared with the blank group,the protein expression of RAGE and TNF-αin the model group increased（P<0.05）;compared with the model group,the protein expression of RAGE and TNF-αin each treatment group decreased（P<0.05）.The ALMD-H group and the positive control group decreased most significantly（P<0.05）.Moreover,The protein expression levels of HMGB1 and RAGE in liver tissues of mice in the model group were highly positively correlated with the level of ALT in serum,but no correlation with TBIL.Conclusion:1.The HMGB1-RAGE signaling pathway may be involved in the occurrence and development of liver injury caused by isoniazid combined with rifampicin and the levels of HMGB1 and RAGE protein are positively correlated with the severity of liver injury.2.Orazamide can prevent liver injury in mice caused by isoniazid combined with rifampin;the mechanism may be related to down-regulating the expression of HMGB1 and RAGE proteins in liver tissues and inhibiting the secretion of inflammatory factors.