Regulation Of Histone Acetylation On Endoplasmic Reticulum Stress In Anti-tuberculosis Drug-induced Liver Injury

Objectives The analysis of the level of GRP78 expression in the serum of patients with antituberculosis drug-induced liver injury and non-patients in the population experiment,and the relationship between GRP78 and liver function indexes of ALT,AST,and verified in cell experiments,to explore the regulatory role of histone acetylation on endoplasmic reticulum stress in ADLI.Methods This subject is mainly studied from two aspects of population and cell.(1)population study: the clinical data and anticoagulant specimens of hospitalized patients who were diagnosed with tuberculosis from July 2016 to December 2016 at the fourth hospital in Tangshan City were collected.63 eligible patients were included in the liver injury group and matched 63 patients as non-liver injury group.ELISA was used to detect the level of GRP78 expression in the patient's plasma.Two independent samples t test was used to analyze the difference of GRP78 expression between the liver injury group and the non-liver injury group,and the correlation between GRP78 and ALT and AST was analyzed.(2)The cytological experiments: firstly,anti tuberculosis drugs are divided into five dosing group,such as INH group,RFP group,PZA group,two-drugs group and three-drugs group,to determine the optimal concentration of CCK8 by cell morphology and liver function index to determine the level of liver injury model;secondly,toxicity detection using CCK8 method is not the same the concentration of C646 and MS-275 in HL-7702 cells,to determine the optimal concentration of the two inhibitors;finally,each dosing group was divided into control group,drug group,C646 group,drug+C646 group,MS-275 group,drug+MS-275 group,to observe the morphology of endoplasmic reticulum cells in each drug group with laser confocal microscopy,the relative expression amount RT-PCR assay of the drug group in P300,HDAC1,GRP78,CHOP m RNA and Western Blot methods to detect the protein content of each drug group;cell culture of each drug group expression of ALT and AST in supernatant;Annexin V/PI double staining method and flow cytometry to detect the cell apoptosis rate in different drug groups.Results The results of this study are divided into two parts.1 the results of crowd research: compared with patients with non-liver injury,the expression of GRP78 in serum of ADLI patients is increased(P<0.05),and GRP78 is positively correlated with ALT and AST(r=0.468,0.386,P <0.05).2 The cytology test results: compared with control group,drug groups decreased endoplasmic reticulum fluorescence intensity,cytoplasmic concentration,HDAC1,GRP78,CHOP m RNA and protein expression were significantly increased(P<0.05),P300 m RNA and protein content decreased significantly(P<0.05),ALT and AST levels in cell culture supernatant increased significantly(P<0.05)and the apoptosis rate increased significantly(P<0.05);compared with the drug groups,drug +C646 groups of endoplasmic reticulum fluorescence intensity changes is not obvious,P300,GRP78 m RNA expression and protein content were significantly decreased(P<0.05),the expression level of CHOP increased significantly(P<0.05),but no significant difference in HDAC1.ALT,AST and cell apoptosis level increased again the rate of cell culture supernatant(P<0.05).But compared with the drug groups,drug+MS-275 groups' HDAC1,CHOP m RNA and protein levels were significantly lower(P<0.05),the expression of GRP78 was significantly increased(P<0.05),but P300 did not change significantly.The fluorescence intensity of endoplasmic reticulum decreased more significantly.The level of ALT and AST and the rate of apoptosis in cell culture supernatant decreased(P<0.05).Conclusions 1 The expression of GRP78 in plasma of ADLI patients was significantly up-regulated,and the level of GRP78 was positively correlated with ALT and AST.2 The changes in histone acetylation and the occurrence of ERS in ADLI process were verified in cells,and the changes in histone acetylation were further confirmed to be involved in the regulation of ERS and then the damage and apoptosis of liver cells.