Competitive Expression Of Circ₀077692 And HDAC2 Influences Antituberculous Drug-induced Liver Injury Through Histone Deacetylation

Objectives Using the circRNA chip in anti-tuberculosis drug-induced liver injury and non-patients,as well as ADLI cell model,were screened to use HDAC2 as the parent gene for differential expression of circRNA,and to verify the common differential expression of circ₀077692 in population and cell model,and to analyze the effect of competitive expression of circ₀077692 and HDAC2 on ADLI through histone deacetylation.Methods This study is divided into two parts: population experiment and cytology experiment.1 population study: 849 cases of TB patients diagnosed in the fifth hospital of Shijiazhuang city were collected,75 cases of non ADLI group and 75 cases of ADLI group were screened after strictly following the inclusion exclusion criteria and matching according to age and gender.The differential expression of circ₀077692 with HDAC2 as parent gene was found by bioinformatics software.The expression levels of pre HDAC2,circ₀077692 and HDAC2 mRNA were detected by RT-PCR,and the expression changes of ALT,AST,SOD,CAT,Bcl-2 and Bax protein were detected by ELISA.2 cytology experiment: to construct ADLI cell model,HL-7702 cells were divided into control group,three drug group,over expressed circrna group and over expressed negative control group by constructing recombinant plasmid over expression technology of circ₀077692.Firstly,CCK8 method was used to determine the optimal concentration and time of action of the drug,and cytological experiments were carried out under this condition.Then he staining was used to observe the changes of cell morphology,the contents of ALT and AST were detected by Lai's method,the level of ROS oxidative stress injury and the rate of apoptosis were detected by flow cytometry,and the mRNA expressions of pre-HDAC2,circ₀077692,HDAC2,Nrf2,SOD,CAT,Bcl-2 and Bax were detected by RT-PCR The expression levels of HDAC2 and Nrf2 were detected by Western Blot,SOD,CAT,Bcl-2 and Bax by ELISA,and the acetylation level of Nrf2 by immunocoprecipitation.3 Statistical analysis: Using spss22.0 software,and using T test and ?² test were used to analyze whether there were differences between the non ADLI group and ADLI group,and Pearson correlation was used to analyze the correlation between the relevant indicators.Results1 population study: there was no significant change in the serum pre-HDAC2 expression in ADLI group compared with that in non ADLI group?P>0.05?The expression level of circ₀077692 was positively correlated with SOD,CAT and Bcl-2,and negatively correlated with ALT,AST and Bax,but HDAC2 The level of mRNA expression was negatively correlated with SOD,CAT and Bcl-2,positively correlated with ALT,AST and Bax?all P<0.05?,and the change trend was opposite to circ₀077692.Two Cytology experiment: CCK8 method determined that the best concentration of drug action of ADLI cell model was 50 ?g/mL+100 ?g/mL+250 ?g/mL?INH+RFP+PZA?;compared with the control group,the morphological changes of the three drug groups showed that the cell damage was serious,ALT and AST contents were increased,ROS oxidative stress injury was aggravated,and apoptosis rate was increased,suggesting that antituberculosis drug stimulation caused damage to human liver cells?P<0.05?.There was no significant change in pre HDAC2 expression?P>0.05?.The expression of circ₀077692,SOD,CAT and Bcl-2 decreased,and the expression of HDAC2 and Bax mRNA increased.The changes were consistent with the results of human experiment.The expression of HDAC2,SOD,CAT,Bcl-2 and Bax protein was consistent with the change of mRNA expression.Nrf2 The expression of mRNA and protein decreased,and the acetylation level of Nrf2 decreased?P<0.05?,while the up-expression of circ₀077692 alleviatedthe damage of hepatocytes?P<0.05?.Conclusions 1 In the serum of patients with ADLI,there was a competitive expression between circ₀077692 and HDAC2 mRNA.circ₀077692 was positively correlated with SOD,CAT and Bcl-2,and negatively correlated with ALT,AST and Bax.HDAC2 mRNA was negatively correlated with ALT,AST,SOD,CAT,Bcl-2 and Bax.2 circ₀077692 reducedadli ADLI by competitive expression with HDAC2 mRNA,which is realized by inhibiting the expression of parent gene HDAC2 enzyme,and then inhibiting the deacetylation of HDAC2,increasing the acetylation level of Nrf2,increasing the level of Nrf2 protein,enhancing the role of antioxidant stress pathway and reducing cell apoptosis.Figure13;Table10;Reference 160...