Differential Expression Of Hsa＿circ＿0082152 In Serum Of Anti-tuberculosis Drug-induced Liver Injury Patients And Its Regulatory Effect For Inflammatory Factors

Objectives Based on previous detection of circRNAs expression profiling in anti-tuberculosis drug-induced liver injury(ADLI)patients and non-ADLI patients and ADLI cell models,this study screened and verified a differentially expressed circRNA:hsa＿circ＿0082152,and explored the mechanism of its regulation on inflammatory factors via a population research and cytological experiments.Methods(1)Population study:238 newly inpatients diagnosed with tuberculosis in the Fifth Hospital of Shijiazhuang City,Hebei Province from August 2020 to December 2020were selected and divided into injury group(108 cases)and non-injury group(130 cases).(2)Cytological experiments:ADLI cell model was established by combined use of anti-tuberculosis drugs.They were divided into control group,drug group,,hsa＿circ＿0082152overexpression and knockout group,metadherin knockout group.HE staining was used to observe the pathological changes of liver cells;RNase R and actinomycin D assays were used to observe the stability of hsa＿circ＿0082152;RPISeq predicts the interaction between hsa＿circ＿0082152 and metadherin(MTDH).RNA-binding protein immunoprecipitation,RNA fluorescence in situ hybridization+immunofluorescence to verify the binding effect of the hsa＿circ＿0082152 and MTDH.(3)Index detection:q RT-PCR was used to detect the m RNA levels of hsa＿circ＿0082152,MTDH,NF-κB,IL-1βand TNF-α;Enzyme linked immunosorbent assay was used to detect the protein levels of ALT,AST and NF-κB,IL-1βand TNF-α;The protein levels of NF-κB and MTDH in each group were detected by Western blot;(4)Statistical analysis:t test,χ~2test,and variance analysis were used to determine the differences between groups;Pearson correlation was used to analyze the correlation between the indicators.Results(1)Population study:the expression of hsa＿circ＿0082152 in the serum of patients with liver injury was higher than that in the non-liver injury group(P<0.05),and liver function indexes(ALT,AST,ALP,TBIL),inflammatory indexes(CRP,LDH))and the expression levels of inflammatory factors(NF-κB,IL-1β,TNF-α)were also up-regulated(all P<0.05);hsa＿circ＿0082152 was positively correlated with liver function indexes,inflammatory indexes and inflammatory factors(all P<0.05).(2)Cytological experiments:hsa＿circ＿0082152 has high stability and mainly exists in the cytoplasm,and its expression is up-regulated in the drug group.The hepatocytes in the drug group were significantly damaged compared with the control group,and the contents of ALT and AST as well as NF-κB,IL-1βand TNF-αwere significantly increased(P<0.05).Compared with the drug group,overexpression of the hsa＿circ＿0082152 can promote the inflammatory response in the process of liver injury,and knockdown of the hsa＿circ＿0082152 can inhibit the inflammatory response in the process of liver injury.hsa＿circ＿0082152 can increase the stability of NF-κB m RNA by binding to MTDH,thereby regulating the inflammatory response during ADLI.Conclusions(1)The hsa＿circ＿0082152 is significantly highly expressed in the serum of patients with liver injury,and is positively correlated with liver function indexes,inflammatory indexes and inflammatory factors,suggesting that hsa＿circ＿0082152 may affect the occurrence of ADLI by regulating the NF-κB inflammatory pathway.(2)The hsa＿circ＿0082152 may positively regulate the NF-κB signaling pathway by binding to MTDH,thereby promoting the inflammatory response during ADLI.Figure 16;Table 12;Reference 157...