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.

中文翻译：

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β-乳球蛋白过敏小鼠模型结肠中改变的 miRNA 谱分析

目标

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

方法

本研究进行了 Illumina 测序，以表征响应 β-Lg 挑战的小鼠结肠中 miRNA 谱的变化。目标基因通过TargetScan 50 和miRanda 3.3a 算法进行预测，并通过GO 和KEGG 分析进行评估。通过细胞转染和定量 RT-PCR 验证所选 miRNA 的表达水平和细胞因子的产生。

结果

共有 15 种 miRNA 在正常小鼠和 β-Lg 致敏小鼠的结肠之间不同表达（P  < 0.05，倍数变化 > 1.50 或 < 0.67），其中包括 6 个上调的 miRNA 和 9 个下调的 miRNA。使用 qRT-PCR 验证了其中的七个 miRNA。GO富集和KEGG通路分析进一步揭示了生物过程、蛋白质结合、细胞质和癌症通路的显着富集，这与过敏性炎症的发展密切相关。此外，在 miRNA 预测算法中鉴定了 β-Lg 过敏中的六个关键功能相互作用对，并使用 qRT-PCR 进行了验证。

结论

我们可以得出结论，我们的结果表明 miRNA 调控网络参与了牛奶过敏的发病机制。