Research On The Expression Of Hsa＿circ＿0093884 In Serum Of Anti-Tuberculosis Drug-induced Liver Injury Patients And Its Related Mechanism

Objectives This study aimed to analyzed the expression of hsa＿circ＿0093884 in serum of anti-tuberculosis drug-induced liver injury(ADLI)patients and non-patients(nonADLI),and to explore the mechanism of hsa＿circ＿0093884 in cell experiments.Methods(1)Subjects: 119 ADLI patients and 133 non-ADLI patients were selected from newly treated patients diagnosed with pulmonary tuberculosis in The Fifth Hospital of Shijiazhuang from August 2020 to January 2021,and their basic clinical information and serum samples were collected.(2)Vitro experiment: Isoniazid,rifampicin and pyrazinamide were used to construct ADLI cell model,which was divided control group,model group,LV-hsa＿circ＿0093884 group,si-hsa＿circ＿0093884 group,etc.Cell morphology was observed by HE staining.The stability of hsa＿circ＿0093884 was detected after treatment with linear digestive enzyme RNase R and actinomycin D.RPISeq and PRIdictor predicted the binding probability of hsa＿circ＿0093884 and RNA-binding protein RPS3.RNA-binding protein immunoprecipitation(RIP)and immunofluorescence assay(IF)were used to confirm it.(3)Index detection: q RT-PCR was used to verify m RNA expression changes of hsa＿circ＿0093884,RPS3,SIRT1,NLRP3,IL-1β and caspase-1;The expression levels of ALT,AST,NLRP3,IL-1β and caspase-1 were detected by ELISA.Western Blot was used to detect the protein expression levels of RPS3 and SIRT1.(4)Statistical analysis: T-test analysis and ANOVA were used for the expression differences of related indicators in serum medium and cell tests of patients in the two groups;Pearson correlation analysis was used to determine the correlation between hsa＿circ＿0093884 and other indicators.Results(1)The expression of hsa＿circ＿0093884 was down-regulated in serum of ADLI patients(P<0.05),and the liver function indexes(ALT,AST,ALP,TBIL),inflammatory indexes(CRP,LDH),NLRP3,IL-1β,caspase-1 were increased(all P<0.05).hsa＿circ＿0093884 was negatively correlated with the liver function indexes,inflammatory indexes,and was positively correlated with SIRT1 expression(all P<0.05).(2)Cell assay:Compared with control group,the cell damage in the model group was obvious,and ALT,AST,NLRP3,IL-1β and caspase-1 were significantly damaged in model group.hsa＿circ＿0093884 and RPS3 co-located in the cytoplasm and could bind to each other.hsa＿circ＿0093884 binding with RPS3 positively regulated SIRT1 and activated SIRT1’s anti-inflammatory effect in ADLI.hsa＿circ＿0093884 weakened the regulation of SIRT1 after RPS3 was knocked down.Conclusions The hsa＿circ＿0093884 was significantly lowly expressed in serum of ADLI patients,and was positively related to SIRT1 and negatively related NLRP3 inflammatory pathways.hsa＿circ＿0093884 competitively activates SIRT1 by binding to RNA binding protein RPS3,thereby regulating hepatocyte inflammation.Figure14;Table 12;Reference 177...