Dong Wenjing, Geng Shang, Cui Junxia, Gao Wenya, Sun Yuena, Xu Tianjun
Laboratory of Fish Molecular Immunology, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China.
Laboratory of Fish Molecular Immunology, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China; National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, China; Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources (Shanghai Ocean University), Ministry of Education, China.
Fish Shellfish Immunol. 2022 Apr;123:94-101. doi: 10.1016/j.fsi.2022.02.043. Epub 2022 Feb 28.
Accumulating evidence has demonstrated that microRNAs (miRNAs) regulate various physiological and pathological processes at the transcriptional level, thus called novel regulators in immune response. In this study, we used bioinformatics and functional experiments to determine the role of miR-103 and miR-190 in the regulation of IL-1R1 gene involved in the immune and inflammatory responses in miiuy croakers. First, we predicted the target genes of miR-103 and miR-190 through bioinformatics and found that IL-1R1 is a direct target gene of miR-103 and miR-190. This was further confirmed by the dual-luciferase reporter assay that the over-expression of miR-103, miR-190 mimics and the pre-miR-103, pre-miR-190 plasmids inhibit the luciferase levels of the wild-type of IL-1R1 3'UTR. miR-103 and miR-190 inhibitors increase the luciferase levels of IL-1R1-3'UTR. Additionally, we found that miR-103 and miR-190 could negatively regulate the mRNA expression of IL-1R1. Importantly, we demonstrated that miR-103 and miR-190 significantly inhibit the NF-κB signaling pathway by targeting IL-1R1 upon LPS stimulation. Collectively, these results provide strong evidence for an important regulatory mechanism of miR-103 and miR-190 targeting the IL-1R1 gene, thereby preventing excessive inflammatory immune responses from causing autoimmunity.
越来越多的证据表明，微小RNA（miRNA）在转录水平上调节各种生理和病理过程，因此被称为免疫反应中的新型调节因子。在本研究中，我们运用生物信息学和功能实验来确定miR-103和miR-190在调节参与大黄鱼免疫和炎症反应的IL-1R1基因中的作用。首先，我们通过生物信息学预测了miR-103和miR-190的靶基因，发现IL-1R1是miR-103和miR-190的直接靶基因。双荧光素酶报告基因检测进一步证实，miR-103、miR-190模拟物以及pre-miR-103、pre-miR-190质粒的过表达会抑制野生型IL-1R1 3'UTR的荧光素酶水平。miR-103和miR-190抑制剂则会提高IL-1R1-3'UTR的荧光素酶水平。此外，我们发现miR-103和miR-190可负向调节IL-1R1的mRNA表达。重要的是，我们证明了在LPS刺激下，miR-103和miR-190通过靶向IL-1R1显著抑制NF-κB信号通路。总体而言，这些结果为miR-103和miR-190靶向IL-1R1基因的重要调节机制提供了有力证据，从而防止过度的炎症免疫反应导致自身免疫。
