Study On Differential Expression Of CircRNAs In Antituberculosis Drug-induced Liver Injury

Objectives To study the differential expression of circRNAs in ADLI and search circRNAs that related to ADLI.Methods The study was carried out in two aspects of cells and crowds.In the study on cells,to construct the liver cell damage model,the HL7702 cells were given two medicine combined?isoniazid and rifampicin?and three medicine combined?isoniazid,rifampicin and pyrazinamide?at levels which made the cell survival rate reach 70%-80%.The cell survival rate was detected by CCK8 method.The HL7702 cells were divided into three groups,the two medicine combined group,the three medicine combined group,and the control group.The cells and cell culture supernatant were collected at 3h,6h,12 h,24h,36 h,and 48 h after dosing.Reitman method was used to detect the expression of ALT and AST;total RNA in cells was extracted by trizol method;Gene Chip was used to screen differentially expressed circRNAs;mi Randa-3.3 software was used to predict mi RNA targets of circRNAs;RT-q PCR method was used to validate the differential expression of circRNAs.One-way ANOVA and t test were used to determine whether there is a difference of ALT,AST,and circRNAs among groups.Pearson correlation analysis was used to estimate whether circRNAs correlate with ALT and AST.In the study on populations,clinical data and blood of hospitalized patients who were diagnosed with tuberculosis were collected from July 2015 to July 2016 at Tangshan tuberculosis hospital.Eligible cases were divided into the group with ADLI and the group without ADLI.These cases were used to screen?16 cases in each group?and validate?98 cases in each group?the differentially expressed circRNAs.The group without ADLI matched the group with ADLI in anti-tuberculosis medicine therapeutic schedule,sex,and age.Methods of circ RNA screening,validation,and prediction of mi RNA targets were the same with the experiment in HL7702 hepatocytes.Paired-samples t test and matching chi-square test were used to analyse the clinical data,circ RNA,ALT and AST.Pearson correlation analysis was used to estimate the correlation between circ RNA and ALT,AST.Results 40965 circRNAs differentially expressed in the two medicine combined group,14863 circRNAs were upregulated,26102 circRNAs were downregulated.10248 circRNAs differentially expressed in the three medicine combined group,2320 circRNAs were upregulated,7928 circRNAs were downregulated.6661 circRNAs differentially expressed in the patient group with ADLI,273 circRNAs were upregulated,6388 circRNAs were downregulated.112 circRNAs differentially expressed in the two cell groups and the patient group.The top upregulated circ RNA both in the two medicine combined group and the patient group with ADLI was hsa_circ₀010996;the top upregulated circ RNA in the three medicine combined group was hsa_circ₀033188;the top downregulated circ RNA in the two medicine combined group,the three medicine combined group and the patient group with ADLI were hsa_circ₀012054,hsa_circ₀025088 and hsa_circ₀021214.The predicted results showed that hsa_circ₀021214 has 92 mi RNA targets,while the other four circRNAs all have 100 mi RNA targets.The verification results showed that hsa_circ₀010996 was upregulated in in the two medicine combined group,the three medicine combined group and the patient group with ADLI.The results were consistented with the screening results.Conclusions 40965 and 10248 circRNAs differentially expressed in the two medicine combined group and the three medicine combined group.6661 circRNAs differentially expressed in the patient group with ADLI.112 circRNAs differentially expressed in the two cell groups and the patient group.hsa_circ₀010996 was validated and had a positive correlation both with ALT and AST.Hsa_circ₀010996 closely related to ADLI.