Regulation Of Inflammatory Factors By Histone Deacetylase SIRT1 In Antituberculosis Drug-induced Liver Injury

Objectives To investigate the mechanism of SIRT1 regulating the expression of inflammatory cytokines in ADLI.Methods The study was carried out in two aspects of crowds and cells.In the study on populations,clinical data and blood of hospitalized patients who were diagnosed with tuberculosis were collected from July 2016 to December 2016 at Tangshan tuberculosis hospital.Eligible cases were divided into the group with ADLI and the group without ADLI(63 cases in each group).T test and chi-square test were used to analyse the clinical data,IL-6 and TNF-? in serum.Pearson correlation analysis was used to estimate the correlation between IL-6,TNF-a and ALT,AST.In the study on cells,the HL-7702 cells were divided into five experimental group: INH group,RFP group,PZA group,two-drug group(INH+RFP)and three-drug group(INH+RFP+PZA).Each experimental group was divided into control group,drug group,drug+SRT1720 group,SRT1720 group,drug+EX527 group and EX527 group.The cell survival rate was detected by CCK8.HE staining was observed the changes of cell morphology.The content of ALT,AST were determined by cell supernatant.The mRNA of SIRT1,NF-?B p65,IL-6 and TNF-? were analyzed by RT-PCR.The protein concentration of SIRT1,NF-?B p65 were detected by WB.The protein concentration of IL-6,TNF-? were detected by ELISA.The acetylation of histone H3K9 in IL-6 promoter regions was detected by CHIP.One-way ANOVA were used to determine whether there is a difference of related indicators among groups.Results In the study on populations,the serum levels of IL-6,TNF-? in patients with ADLI were elevated,and were positively correlated with ALT,AST.In the study on cells,the drug concentrations were determined to be 800 ?g/ml INH,200 ?g/ml RFP,400 ?g/ml PZA,75+150 ?g/ml(INH+RFP),and 50+100+267 ?g/ml(INH+RFP+PZA);the combined concentrations of SRT1720 and EX527 were both 1 ?mol/L.After cultured for 48 h,cell morphology suggested that cells in each grug group exhibited varying degrees of apoptotic injury,especially two-drug group and three-drug group.The multiple increase of ALT and AST in two-drug group and three-drug group was higher than that of the drug alone,suggesting that different types of anti-tuberculosis drugs can lead to liver injury,and the combination of drugs is more serious.Combined use of SRT1720 can reduce liver cell damage caused by antituberculosis drugs,and combined use of EX527 further aggravates the occurrence of injury.Compared with the blank control group,the expression of SIRT1 mRNA was decreased after treated with INH,RFP,PZA,two-drugs and three-drugs(P<0.05),the mRNA expression of NF-?B p65,IL-6 and TNF-? in INH group,two-drug group and three drug group was higher than that in the blank control group(P<0.001).The combination of above three groups with SRT1720 could inhibit the increase of NF-?B p65,IL-6 and TNF-? mRNA expression induced by anti-tuberculosis drugs stimulation,suggesting that SIRT1 activation can reduce NF-?B p65 expression.In turn,reduce the expression of IL-6 and TNF-? to reduce the inflammatory response caused by antituberculosis drugs.The combination of these three groups with EX527 will further increase the expression of NF-?B p65,IL-6 and TNF-? mRNA,indicating that SIRT1 inhibition can be increased by increasing the expression of NF-?B p65,TNF-? and IL-6 to increase the inflammatory damage caused by antituberculosis drugs.Related indicators protein expression is consistent with the corresponding mRNA expression.Furthermore,the level of histone H3K9 acetylation in the IL-6 promoter region of INH group cells was higher than that in the blank control group.Combined use of SRT1720,the level of acetylation of histone H3K9 in the promoter region of IL-6 was decreased,and combined with EX527,the acetylation level was further increased.Conclusions The serum levels of IL-6 and TNF-? in patients with ADLI were elevated,and were positively correlated with ALT and AST.The expression level of SIRT1 was decreased and the expression of inflammatory factors was increased during the process of human hepatocyte injury caused by anti-TB drugs.Activation of SIRT1 can reduce the occurrence of hepatocyte injury by reducing the expression of inflammatory regulators.Inhibition of SIRT1 aggravates hepatocyte inflammatory lesions.The mechanism by which SIRT1 participates in Isoniazid-induced liver cell injury may be accomplished by deacetylation of histone H3K9 in IL-6 promoter region.