ABSTRACT

Background: The regulatory network of ulcerative colitis (UC)-associated miRNAs is not fully understood. In this study, we aim to investigate the global profile and regulatory network of UC associated miRNAs in the context of dextran sulfate sodium (DSS). Methods: UC was induced in C57BL mice using DSS. Differentially expressed miRNAs were screened by RNA sequencing and subjected to the Kyoto Encyclopedia of Genes and Genomes Pathway enrichment analysis. RT-qPCR was used to verify the differential expression of miRNAs and candidate target mRNA. Luciferase reporter vector bearing a miRNA binding site was constructed to verify the binding site of the miRNA on mRNA. Results:A total of 95 miRNAs (31 were up-regulated and 64 were down regulated) differentially expressed in the colonic tissues of the UC mice. Among the differentially expressed miRNAs, IL-25 pathway genes were enriched. Subsequent RT-qPCR confirmed a decreased expression of IL-25 and a significant up regulation of IL-25 target miRNAs including mmu-miR-135b-5p, mmu-miR-7239-5p and mmu-miR-691 in UC mice. Conclusion: Using the luciferase assay, we verified the biding sites of mmu-miR-135b-5p and mmu-miR-691 to the IL-25 3ʹUTR. In conclusion, mmu-miR-135b-5p:IL-25 and mmu-miR-691:IL-25 interactions are important pathways that may exert a protective role in UC.

摘要

背景：与溃疡性结肠炎（UC）相关的miRNA的调控网络尚未完全了解。在这项研究中，我们旨在研究硫酸葡聚糖硫酸钠（DSS）背景下UC相关miRNA的全球概况和调控网络。方法：使用DSS在C57BL小鼠中诱发UC。通过RNA测序筛选差异表达的miRNA，并对其进行《京都议定书》基因和基因组途径富集分析。RT-qPCR用于验证miRNA和候选靶mRNA的差异表达。构建带有miRNA结合位点的荧光素酶报告载体，以验证miRNA在mRNA上的结合位点。结果：在UC小鼠结肠组织中共有95个miRNA差异表达（31个上调而64个下调）。在差异表达的miRNA中，IL-25途径基因富集。随后的RT-qPCR证实了UC小鼠中IL-25的表达降低和IL-25靶miRNA的显着上调，包括mmu-miR-135b-5p，mmu-miR-7239-5p和mmu-miR-691。结论：使用荧光素酶测定法，我们验证了mmu-miR-135b-5p和mmu-miR-691对IL-25 3ʹUTR的结合位点。总之，mmu-miR-135b-5p：IL-25和mmu-miR-691：IL-25相互作用是可能在UC中发挥保护作用的重要途径。