长链多不饱和脂肪酸的组合调节了哮喘模型小鼠和人肺泡细胞中 miRNA-146a-5p 的表达。

A combination of LCPUFAs regulates the expression of miRNA-146a-5p in a murine asthma model and human alveolar cells.

机构信息

Department of Food Technology, University of Applied Sciences, Leipziger Str. 123, Fulda, Germany; Division for Allergy, Pneumology and Cystic Fibrosis, Department for Children and Adolescents, Goethe University, Theodor-Stern-Kai 7, Frankfurt/Main, Germany; Faculty of Biological Sciences, Goethe University Frankfurt/Main, Max-von-Laue-Straße 9, Frankfurt/Main, Germany.

Division for Allergy, Pneumology and Cystic Fibrosis, Department for Children and Adolescents, Goethe University, Theodor-Stern-Kai 7, Frankfurt/Main, Germany.

出版信息

Prostaglandins Other Lipid Mediat. 2020 Apr;147:106378. doi: 10.1016/j.prostaglandins.2019.106378. Epub 2019 Nov 4.

DOI:10.1016/j.prostaglandins.2019.106378

PMID:31698144

Abstract

BACKGROUND

LCPUFAs are suggestive of having beneficial effects on inflammatory diseases such as asthma. However, little is known about the modulative capacity of omega-(n)-3 and n-6 LCPUFAs within the epigenetic regulation of inflammatory processes.

OBJECTIVE

The aim of this study was to investigate whether a specific combined LCPUFA supplementation restores disease-dysregulated miRNA-profiles in asthmatic mice. In addition, we determined the effect of the LCPUFA supplementation on the interaction of the most regulated miRNA expression and oxygenase activity in vitro.

METHODS

Sequencing of miRNA was performed by NGS from lung tissue of asthmatic and control mice with normal diet, as well as of LCPUFA supplemented asthmatic mice. Network analysis and evaluation of the biological targets of the miRNAs were performed by DIANA- miRPath v.3 webserver software, TargetScanMouse 7.2, and tool String v.10, respectively. Expression of hsa-miRNA-146a-5p and activity of COX-2 and 5-LO in LCPUFA-treated A549 cells were assessed by qPCR and flow cytometry, respectively.

RESULTS

In total, 62 miRNAs were dysregulated significantly in murine allergic asthma. The LCPUFA combination restored 21 of these dysregulated miRNAs, of which eight (mmu-miR-146a-5p, -30a-3p, -139-5p, -669p-5p, -145a-5p, -669a-5p, -342-3p and -15b-5p) were even normalized compared to the control levels. Interestingly, six of the eight rescued miRNAs are functionally implicated in TGF-β signaling, ECM-receptor interaction and fatty acid biosynthesis. Furthermore, in vitro experiments demonstrated that upregulation of hsa-miRNA-146a-5p is accompanied by a reduction of COX-2 and 5-LO activity. Moreover, transfection experiments revealed that LCPUFAs inhibit 5-LO activity in the presence and absence of anti-miR-146a-5p.

CONCLUSION

Our results demonstrate the modulative capacity of LCPUFAs on dysregulated miRNA expression in asthma. In addition, we pointed out the high regulative potential of LCPUFAs on 5-LO regulation and provided evidence that miR-146a partly controls the regulation of 5-LO.

摘要

背景

长链多不饱和脂肪酸（LCPUFAs）对哮喘等炎症性疾病具有有益影响。然而，人们对 ω-(n)-3 和 n-6 LCPUFAs 在炎症过程的表观遗传调控中的调节能力知之甚少。

目的

本研究旨在探讨特定的联合 LCPUFA 补充是否能恢复哮喘小鼠疾病失调的 miRNA 谱。此外，我们还确定了 LCPUFA 补充对体外最受调节的 miRNA 表达和加氧酶活性相互作用的影响。

方法

通过对正常饮食的哮喘和对照小鼠以及补充 LCPUFA 的哮喘小鼠的肺组织进行 NGS 测序，对 miRNA 进行测序。通过 DIANA- miRPath v.3 网络分析和评估软件、TargetScanMouse 7.2 和 tool String v.10 分别对 miRNA 的生物靶标进行网络分析和评估。通过 qPCR 和流式细胞术分别评估 hsa-miRNA-146a-5p 的表达和 COX-2 和 5-LO 的活性。

结果

总共，62 个 miRNA 在过敏性哮喘的小鼠中显著失调。LCPUFA 联合治疗恢复了 21 个失调的 miRNA，其中 8 个（mmu-miR-146a-5p、-30a-3p、-139-5p、-669p-5p、-145a-5p、-669a-5p、-342-3p 和 -15b-5p）甚至与对照水平正常化。有趣的是，在功能上，这 8 个恢复的 miRNA 中有 6 个与 TGF-β 信号、ECM-受体相互作用和脂肪酸生物合成有关。此外，体外实验表明，hsa-miRNA-146a-5p 的上调伴随着 COX-2 和 5-LO 活性的降低。此外，转染实验表明，LCPUFAs 在存在和不存在抗 miR-146a-5p 的情况下抑制 5-LO 活性。