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miRNA-IL-25 相互作用在溃疡性结肠炎小鼠中的可能作用。

Possible role of microRNA miRNA-IL-25 interaction in mice with ulcerative colitis.

机构信息

Department of Gastroenterology, Jinan University of Second Clinical Medical Sciences, Shenzhen People's Hospital , Shenzhen, Guangdong Province, China.

Department of General Surgery, Shenzhen Children's Hospital , Shenzhen, Guangdong Province, China.

出版信息

Bioengineered. 2020 Dec;11(1):862-871. doi: 10.1080/21655979.2020.1804176.

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 3'UTR. 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

溃疡性结肠炎(UC)相关 miRNA 的调控网络尚未完全阐明。本研究旨在探讨葡聚糖硫酸钠(DSS)诱导的 UC 相关 miRNA 的整体谱和调控网络。

方法

采用 DSS 诱导 C57BL 小鼠 UC。通过 RNA 测序筛选差异表达 miRNA,并进行京都基因与基因组百科全书通路富集分析。采用 RT-qPCR 验证 miRNA 和候选靶 mRNA 的差异表达。构建携带 miRNA 结合位点的荧光素酶报告载体,验证 miRNA 对 mRNA 的结合位点。

结果

UC 小鼠结肠组织中共有 95 个 miRNA (31 个上调,64 个下调)差异表达。差异表达 miRNA 中,IL-25 通路基因富集。随后的 RT-qPCR 证实 UC 小鼠中 IL-25 表达下调,IL-25 靶标 miRNA 包括 mmu-miR-135b-5p、mmu-miR-7239-5p 和 mmu-miR-691 显著上调。

结论

通过荧光素酶检测实验,我们验证了 mmu-miR-135b-5p 和 mmu-miR-691 与 IL-25 3'UTR 的结合位点。综上所述,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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ef0/8291871/87dd7087f897/KBIE_A_1804176_UF0001_OC.jpg

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