β-乳球蛋白过敏小鼠结肠中差异表达 miRNA 谱分析。

Analysis of altered miRNA profiling in the colon of a mouse model with β-lactoglobulin allergy.

机构信息

Key Laboratory of Dairy Science, Ministry of Education, Food College, Northeast Agriculture University, Harbin, China.

出版信息

Allergol Immunopathol (Madr). 2020 Nov-Dec;48(6):666-674. doi: 10.1016/j.aller.2020.05.007. Epub 2020 Oct 29.

DOI:10.1016/j.aller.2020.05.007

PMID:33131977

Abstract

OBJECTIVES

The differences in the expression profiles of colonic miRNAs between β-lactoglobulin (β-Lg) allergic mice and normal mice were analyzed to investigate the important role of the miRNA regulation mechanism in the pathogenesis of cow's milk allergy.

METHODS

The present study performed Illumina sequencing to characterize the miRNA profile changes in mouse colon responding to β-Lg challenge. Target genes were predicted by TargetScan 50 and miRanda 3.3a algorithms and assessed by GO and KEGG analysis. The expression levels of selected miRNAs and cytokine production were verified by cell transfection and quantitative RT-PCR.

RESULTS

A total of 15 miRNAs were diversely expressed between the colon of the normal and β-Lg-sensitized mice (P < 0.05, fold change of >1.50 or <0.67), including six up-regulated miRNAs and nine down-regulated miRNAs, among which seven miRNAs were validated using qRT-PCR. GO enrichment and KEGG pathway analyses further revealed that biological process, protein binding, cytoplasm and the pathways of cancer were significantly enriched, which were closely connected to the allergic inflammation development. Additionally, six key functional interaction pairs in β-Lg allergy were identified in miRNA prediction algorithms and verified using qRT-PCR.

CONCLUSIONS

We can conclude that our results suggested that the miRNAs regulation network participated in the pathogenesis of cow's milk allergy.

摘要

目的

分析β-乳球蛋白（β-Lg）过敏小鼠和正常小鼠结肠中 miRNA 表达谱的差异，以探讨 miRNA 调控机制在牛奶过敏发病机制中的重要作用。

方法

本研究通过 Illumina 测序对 β-Lg 刺激后小鼠结肠中 miRNA 谱的变化进行了特征分析。利用 TargetScan 50 和 miRanda 3.3a 算法预测靶基因，并进行 GO 和 KEGG 分析。通过细胞转染和定量 RT-PCR 验证所选 miRNA 和细胞因子产生的表达水平。

结果

正常和β-Lg 致敏小鼠结肠之间有 15 个 miRNA 表达存在差异（P<0.05，倍数变化>1.50 或<0.67），包括 6 个上调 miRNA 和 9 个下调 miRNA，其中 7 个 miRNA 通过 qRT-PCR 进行了验证。GO 富集和 KEGG 通路分析进一步表明，生物过程、蛋白结合、细胞质和癌症通路显著富集，这与过敏炎症的发展密切相关。此外，通过 miRNA 预测算法鉴定到了 6 个关键的功能性 β-Lg 过敏互作对，并通过 qRT-PCR 进行了验证。