The Effect Of HMGB1-RAGE Signal Pathway In Liver Injury Induced By Rifampicin And Its Possible Mechanism

Objective: rifampicin alone or in combination with other antituberculosis drugs can cause different degrees of liver damage,but the specific pathogenesis is not yet clear.The high mobility group protein B1 is an inflammatory cytokine found in recent years,which is related to the development of liver injury in rats with isoniazid combined with rifampicin.Objective: 1)to replicate the liver injury model induced by rifampin,and to observe the effect of HMGB1 release inhibitor,ethyl pyruvate(EP)on the model,and to explore its possible mechanism.2)use HMGB1 polyclonal antibody and RAGE monoclonal antibody to block HMGB1-RAGE signal transduction pathway,and observe its effect on liver damage model mice induced by rifampin.Methods: 1)the 24 SFP C57BL/6 mice were randomly divided into 3 groups,namely,normal group,RFP group and EP group,8 each group.The mice in the RFP group and EP group were given 200 mg/ kg(kg·d)per day,and the normal group gave the same amount of sodium carboxymethyl cellulose to the stomach.In the EP group,the mice were injected with ethyl pyruvate 40 mg/(kg·d)per day,and the other two groups were given the same amount of green's intraperitoneal injection.Building 7 days executed after mice,detection of mice serum ALT,TBIL,DBIL,ALP,observe the mice liver pathology change,detection of mice liver tissue associates,AC-HMGB1 level,determine the expression of liver tissue MDA and SOD,detection of HMGB1 expression in the organization.2)randomly divided 40 C57BL/ 6 SFP mice into 5 groups,namely,normal group,RFP group,HMGB1 antibody intervention group,RAGE antibody intervention group and IgG control group,each group of 8.In addition to the normal group,the mice ineach group were given a daily dose of 200 mg/(kg·d).The HMGB1 antibody intervention group was injected with HMGB1 polyclonal antibody 5mg/Kg in the first half hour after the first,third and fifth days of the molds.The rrage antibody intervention group was injected into the first half hour of the molds in the 1st,3rd,and5 th days of the molds,and the first half hour of intraperitoneal injection of RAGE monoclonal antibody was 0.5mg/Kg.IgG control group was injected with rat IgG5mg/Kg in the first half hour of the molds in the first,third and fifth days of the molds.In the RFP group,the mice were injected with the same amount of solvent as the control.Mouse serum TBIL,DBIL,ALP,ALT,TBA level were detected in the 7 days after the model was created,and the histopathological changes of liver tissues were observed to detect the level of HMGB1 protein expression in liver tissue.Results: 1)compared with normal group,serum TBIL,DBIL,ALP and ALT were significantly increased in RFP group(P < 0.05).The hepatic cells showed significant vacuolar degeneration and fatty degeneration,and local hepatocytes were seen to be eosinophilic,nucleated,or dissolved.The content of TBA and MDA in liver tissues increased(P < 0.05),SOD decreased(P < 0.05),and the expression of HMGB1 and ac-hmgb1 increased(P < 0.05),and HMGB1 cytoplasmic expression was the main expression.Compared with the RFP group,the serum TBIL,DBIL,ALP and ALT levels in the EP group were decreased(P < 0.05).Hepatic histopathological changes;The content of TBA and MDA in liver tissues decreased(P < 0.05),SOD increased(P <0.05),and the expression level of HMGB1 and ac-hmgb1 decreased(P < 0.05),and HMGB1 cell nuclear expression was the main expression.2)compared with normal group,serum TBIL,DBIL,ALP,ALT and TBA in RFP group were increased(P < 0.05).The vacuolar degeneration and fatty degeneration of hepatocyte are obvious,and partial liver cells can be seen nucleolysis or consolidation,locally visible point necrosis and inflammatory cell infiltration.The expression of HMGB1?RAGE?NF-kB in liver tissues improved significantly(P < 0.05),and theexpression of BSEP decreased significantly(P < 0.05)..Compared with the RFP group,the above indexes in the IgG control group showed no significant difference((P < 0.05)).The levels of serum TBIL,DBIL,ALP,ALT and TBA in mice with HMGB1 antibody intervention group and RAGE antibody intervention group were decreased(P < 0.05),and the pathological changes of liver tissue were improved.The expression of HMGB1?RAGE?NF-kB in liver tissues decreased significantly(P < 0.05),and the expression of BSEP improved significantly(P < 0.05).Compared with the IgG control group,the intervention group of HMGB1 antibody and the intervention group of RAGE antibody were significantly improved(P < 0.05).There was no significant difference between the HMGB1 antibody intervention group and the RAGE antibody intervention group(P >0.05).Conclusions:1.The distribution and expression intensity of HMGB1 are basically consistent with the pathological changes of the liver,indicating that the increase of HMGB1 expression is related to the progression of the abnormal distribution of HMGB1 and the increase of ac-hmgb1 in liver injury induced by rifuping.2.Ethyl pyruvate can reduce the level of HMGB1 and its acetylation level and regulate the distribution of HMGB1 to reduce the drug-induced liver injury of rifampicin through antioxidant.3.Blockade of HMGB1-RAGE signaling pathway can significantly reduce the severity of liver injury induced by rifampicin in mice,and reduce the expression of liver HMGB1,RAGE,NF-kB,suggesting that HMGB1-RAGE-NF-kB signaling pathway is involved in rifampicin Induced liver damage progression.