Effect Of Histone Acetylation On Expression Of CYP2E1 In Antituberculosis Drug-induced Liver Injury

Objectives Histone acetylation alters the effect of CYP2E1 expression differences in anti-tuberculosis drug-induced liver injury.Methods It mainly studies from both the cell and the population.In the population study,the clinical data and blood specimens of newly diagnosed patients diagnosed by the Tuberculosis Hospital of Tangshan from July 2016 to December 2016 were collected.The eligible cases were included in the liver injury group and the non-liver injury group,respectively.Elisa method was used to detect the CYP2E1 level in the patient's serum.The use of Chi-square test or t-test to determine whether the population data,CYP2E1 content,ALT and AST have differences between groups;using Pearson correlation analysis to determine whether there is a correlation between CYP2E1 content and ALT,AST.In cell studies,isoniazid,rifampicin,pyrazinamide,a combination of two drugs,and three drugs were combined and diluted to different concentration gradients to effect hepatocytes for 24 hours.The drug concentration was selected to be 80-85%.After determination of drug concentration,CYP2E1 mRNA was detected at 6,24,and 48 hours after dosing,and CYP2E1 mRNA was increased at 6 hours,so the dosing time was determined to be 6 hours.Colorimetric assay was used to detect the activity of ALT and AST in cell culture medium;HE staining was used to observe the cell morphology;Elisa method was used to detect hepatocyte histone acetylase(HAT)and histone deacetylase.(HDAC)activity and JNK protein content;detection of acetylated histone H3K56,H4K5 and CYP2E1 protein levels by Western blot;qRT-PCR detection of CYP2E1 mRNA,JNK mRNA expression changes;ChIP method detection of CYP2E1 gene initiation The acetylation levels of histone H3K56 and H4K5 changed.Results Population experiment: Compared with the level of CYP2E1 protein in the serum of non-hepatic injury patients,the expression level of CYP2E1 protein in serum of ADLI patients was increased,the difference was statistically significant(P <0.05),and was positively correlated with the liver function indicators ALT,AST.Part of the cells: Compared with the control group,the HAT activity in the isoniazid group was decreased,HDAC activity was increased,acetylated H3K56 and H4K5 were decreased(P <0.05),and HAT activity was decreased in the combination of rifampicin,the two drugs,and the three drugs.The activity of HDAC increased,the activity of pyrazinamide HAT remained unchanged,and the activity of HDAC increased.The acetylated H3K56 and H4K5 were increased in all drug groups,the mRNA and protein of CYP2E1 and JNK were increased,and the ALT and AST activities were increased(P <0.05).The cells showed long spindles and triangles,and the number decreased.Compared with the drug group,the HAT activity in the drug plus Garcinol group decreased,the acetylated H3K56,H4K5 decreased,the HDAC activity in the drug plus TSA group decreased,and the acetylated H3K56,H4K5 increased.In the drug plus inhibitor group,the mRNA and protein levels of CYP2E1 and JNK decreased,and the ALT and AST activities decreased(P <0.05).Long spindle cells decreased;compared with the control group,the HAT activity in the Garcinol group decreased,and the HDAC activity in the TSA group decreased.Decrease(P <0.05),no difference in other indicators.Comparing the results of drug groups,HAT activity was increased,HDAC activity was decreased,acetylated H3K56 was increased,mRNA and protein of CYP2E1,JNK were increased,and ALT,AST were increased in the two-drug combination group and the three-drug combination group compared with each drug group.Increased activity.(P<0.05).Conclusions CYP2E1 protein in plasma of patients with tuberculosis is closely related to ADLI.The expression of CYP2E1 protein is significantly up-regulated in plasma of ADLI patients.The mechanism is related to the change of histone acetylation caused by antituberculosis drugs.