The Filtering Of MicroRNAs Related Hepatic Cells Regeneration After HBA-related Acute-on-chronic Liver Failure

Objective:(1) To obtain the differential expression profiles of plasma miRNA in HBV-related acute-on-chronic liver failure(ACLF) patients between different groups and intra-group after the disease evolution.(2) To screen out the miRNA and their target genes which relate to regeneration of hepatic cells after ACLF and set a theoretical basis for the further research of miRNA' function and mechanism in hepatic cells regeneration after HBV-related ACLF.Methods: The plasma samples of early stage HBV related ACLF patients are collected in the infectious diseases department of the affiliated hospital of Southwest Medical University. If their disease had no improvement or progressed to end-stage, their plasma samples would be collected once again. If their disease improved and their liver function turned to normal, their plasma samples would be collected once again. All the patients will be followed up for at least 24 weeks. Their clinical data and demographic data will be collected at the same time. These ACLF patients are divided into survival group(GS) and death group(Gd) based on their clinical outcomes. We choose 3 patients from each group and collect their plasma samples at two time points according to the median of every indicator of these two groups. We analysis the miRNA expressing differences between two groups and intra-group after disease progress using LNA-miRNA microarray then screen out the miRNA according to results(up-regulate or down-regulate more than 1.5 times and P<0.05). Then we find out the miRNA and their target genes, related to cell proliferation or/and hepatocyte proliferation and apoptosis, by using the databases for bioinformatics analysis(miRBase, Pub Med and Medline).Results: Comparing miRNA expression levels between baseline(first diagnosis) and follow-up in GS patients, the expression level of 41 miRNA changed significantly; 27 miRNA(hsa-miR-4468, hsa-miR-3940-5p, hsa-miR-4732-5p, hsa-miR-200c-3p, hsa-miR-541-5p,hsa-miR-660-3p, hsa-miR-4777-5p, etc.) up-regulated. 14 miRNA(hsa-miR-30b-5p, hsa-let-7f-2-3p, hsa-miR-1297, hsa-miR-548 y,hsa-miR-4451, etc.) down-regulated. Comparing miRNA expression levels at baseline between GS and Gd, the expression level of 57 miRNA changed significantly, 35 miRNA(hsa-miR-491-3p, hsa-miR-27b-3p, hsa-miR-4532, hsa-miR-125a-5p, hsa-miR-4456, hsa-miR-222-3p, hsa-miR-298, hsa-miR-222-3p, hsa-miR-3940-5p, etc.)up-regulated. 22 miRNA(hsa-miR-146b-5p, hsa-miR-1297, hsa-miR-142-5p, hsa-miR-223-3p, hsa-let-7f-2-3p, hsa-miR-4739, etc.)down-regulated. Comparing miRNA expression levels between the baseline(first diagnosis) and the followed-up in Gd patients, we found that the expression level of 8 miRNA changed significantly, two miRNA including hsa-miR-142-3p and hsa-miR-1827 up-regulated. Six miRNA down-regulated including hsa-miR-875-3p, hsa-miR-4521,hsa-miR-5579-3p, hsa-miR-641, hsa-miR-548a-5p, hsa-miR-4796-3p.The result of bioinformatics analysis implied that 10 miRNA(hsa-miR-148a-3p, hsa-miR-1297, hsa-miR-142a-3p, hsa-miR-125a-5p, etc.)were associated with regeneration of hepatic cells and their functions involved in the regulation of all kinds of cell proliferation and apoptosis;34 target genes(EGFR, SMAD4, CXCR4, PTEN, etc.) were associated with regeneration of hepatic cells and their functions involved in the regulation of all kinds of cell proliferation and apoptosis;Conclusions:(1)The differentially expressed miRNA in plasma may be involved in the process of HBV-related ACLF with different clinical outcomes.(2)10 miRNA(hsa-miR-148a-3p, hsa-miR-1297, hsa-miR-142a-3p,hsa-miR-125a-5p, etc.) may participate in the process of regeneration of hepatic cells after ACLF, while its specific mechanism needs to be studied further.