| BackgroudPrimary biliary cirrhosis(PBC) is a chronic liver disease characterised by intrahepatic bile-duct destruction, cholestasis, and, in some cases, cirrhosis. Evidence supporting the autoimmune nature of this disorder includes the appearance of highly specific antimitochondrial antibodies (AMAs) and autoreactive T cells. Study of pathogenesis will help to improve the level of clinical diagnosis and treatment. MicroRNAs (miRNA) are small, single-stranded noncoding RNAs, which is demonstrated to suppress the expression of specific genes at the translation level by complementary binding to the3’ noncoding region of the target mRNA. MiRNAs participate in many vital processes, including cell proliferation, cell differentiation and apoptosis. However, the expression profile in peripheral blood mononuclear cells (PBMCs) from PBC patients and the role of microRNA in PBC remained unclear. MiRNA expression profiles have been identified in several autoimmune diseases, at the same time, only specific antimitochondrial antibodies (AMA) cann’t meet the clinical needs for PBC diagnosis.MiRNA is relatively stable and also can be used as new disease biomarkers for diagnosis, classification and prognosis.Our study aimed to explore abnormal microRNA regulation in PBC.Objective This study was undertaken to explore miRNA expression profile in PBMC of patients with PBC, and to identify the miRNAs associated with the pathogenesis of PBC, as well as their relationship with clinical features.This study aim to find out PBC specific miRNAs as a new disease markers.Then through bioinformatics prediction and dual luciferase report gene assay system to study further mechanism about disease.MethodsPBMCs isolated from4patients with PBC and3healthy controls were used for miRNA microarray analysis to detect differentially expressed miRNAs between the two groups.We identified9miRNAs correlated with PBC by bioinformatics analysis.The expression of miR-21、miR-181a、miR-150-5p、miR-155-5p、miR-142-3p、miR-23a、 miR-23b、miR146a and miR-146b in50PBC patients and30healthy controls were measured by a quantitative real-time qPCR assay. The correlation between the expression of miRNAs and the clinical or laboratory features has been investigated. Real time quantitative PCR was also used for detecting the levels of miR-150-5p、miR-155-5p in CD19+B cell and CD4+T cell.Bioinformatics prediction and dual luciferase report gene assay system were performed to identify miR-150target gene.ResultsWe performed a miRNA profiling analysis of4PBC patients and3healthy controls, and selected out12differentially expressed miRNAs with9up-regulated and3down-regulated in PBC. Real-time qPCR confirmed the expression level of miR-155-5p in PBC patients were significantly lower than those in healthy controls, while the expression level of miR-150-5p in PBC patients were significantly higher than those in healthy controls.The most obvious expression level of miR-150-5p was in CD4+T cells.There has no obvious expression between patients with PBC and healthy controls in other miRNAs as showen above. Further analysis showed that IgA was positively correlated correlated with the expression level of miR-150-5p in patient’s PBMCs. No other correlation between clinical or laboratory features with miRNAs was discovered.The expression of miRNAs in different sub-group had no obvious difference. The target genes of miR-150-5p were predicted by bioinformatic method first.We found that miR-150-5p significantly inhibited the luciferase activity of the3UTR reporter.ConclusionOur findings revealed a significant different expression of microRNAs in PBMC between patients with PBC and healthy controls, at while time,we also have preliminary studied the function of this miRNA.These may be related to the pathogenesis of PBC. It can become a new diagnostic biomarker and treatment target. Its pathogenesis needs further studies. |
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