Profile Of Circulating MicroRNAs Expression In HBV Associated Acute-on-chronic Liver Failure Patients And Its Correlation With Prognosis

Objective1) To investigate the profile of circulating miRNA expression inHBV related acute-on-chronic liver failure (ACLF) patients.2) To study the correlationbetween the miRNAs and prognosis in ACLF patients.3) To discuss the target genes ofmiR-142and their correlation in ACLF patients. Methods The serum of6ACLF patientsand6healthly controls (HC) were collected in Ruijin hospital. MiRNAs, isolated from theserum, were detected by microarray chip. Basing on the result of microarray chip,miRNAs with significant changes were selected. Patients hospitalized in Ruijin hospitalwere enrolled, including20chronic hepatitis B (CHB),20HBV related liver cirrhosis(LC),50ACLF and40HCs. Serum miRNAs were detected by real-time PCR. Thefrequency of Th17cells and the expression of IL-17in peripheral blood were detected byflow cytometry or Enzyme-linked immmunosorbent Assay (ELISA). In addition, the targetgenes of miR-142were analyzed by bioinformatics, including GO and KEGG. Thecorrelations between serum miRNAs and target genes were analyzed. One way ANOVAtest and t test were used to compare the differences among groups and the correlation wereanalyzed by Pearson and Spearman correlation depengding on date distribution. ResultsThe analysis of locked nucleic acid (LNA)-microRNAs microarray showed that92miRNAs changed significantly, including10increased miRNAs and82decreasedmiRNAs in ACLF patients. Compared with HCs, the expression of serum hsa-miR-595,hsa-miR-300and hsa-miR-491-5p were increased in CHB, LC and ACLF patients,especially in ACLF group (fold change=5.63, p <0.001; fold change=2.14, p <0.001; foldchange=3.23, p <0.001). Meanwhile, the levels of hsa-miR-130a, hsa-miR-200b, hsa-miR-142-3p and hsa-miR-142-5p in CHB, LC and ACLF patients were decreasedmarkedly compared with HCs, especially in ACLF group (fold change=0.25, p <0.001.fold change=0.24, p <0.001. fold change=0.28, p <0.001. fold change=0.3, p <0.001).There were positive correlations between the expression of hsa-miR-595, hsa-miR-491-5pand MELD score in ACLF group(r=0.8449，p<0.05；r=0.5006，p<0.05). The expressionof hsa-miR-130a, hsa-miR-200b, hsa-miR-142-3p, hsa-miR-142-5p were negativecorrelated with MELD score in ACLF group (r=-0.4470，p<0.05；r=-0.5371，p<0.05；r=-0.4973， p<0.05； r=-0.3539， p<0.05). The levels of serum hsa-miR-595andhsa-miR-300were decreased along with disease recovery (p<0.05), while the expressionof hsa-miR-130-a, and hsa-miR-142-5p were increased compared with baseline in survivalsubjects of ACLF group (p<0.05). However, in the non-survival group, the4miRNAswere changed without significance. The frequency of Th17cells in ACLF patients weresignificantly increased than that of in HCs ((2.88±1.28)%Vs (2.04±0.92)%)(P<0.01).IL-17levels were significantly elevated in ACLF patients compared to HCs((69.89±31.93pg/ml) Vs (49.11±23.72pg/ml))(P=0.01). Bioinformatics analysis (GO andKEGG web) showed that IL-17might be negatively regulated by miR-142. Statisticanalysis indicated that the levels of has-miR-142-3p and has-miR-142-5p were negativelycorrelated with the expression of serum IL-17in ACLF group (r=-0.5859, p=0.006;r=-0.4827, p=0.014). Conclusions In ACLF patients, the expression of hsa-miR-595,hsa-miR-300and hsa-miR-491-5p were upregulated obviously, while hsa-miR-130a,hsa-miR-200b, hsa-miR-142-3p and hsa-miR-142-5p were downregulated significantly.These miRNAs were closely correlated with disease severity and clinical outcomes inACLF.The imbalanced profile of miRNAs might be involved in the pathogenesis ofACLF. Hsa-miR-142-3p and hsa-miR-142-5p may negatively regulate the expression ofIL-17in the process of ACLF. Further study is needed in demonstrating the mechanism.