Guo Jian, Zhang Ruiya, Zhao Yiqing, Wang Junping
Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, People's Republic of China.
Department of General Surgery, Shanxi Provincial People's Hospital, The Affiliated People's Hospital of Shanxi Medical University, Taiyuan, 030012, People's Republic of China.
J Inflamm Res. 2021 May 18;14:2031-2043. doi: 10.2147/JIR.S302832. eCollection 2021.
Dysregulation of micro-RNAs (miRNAs) is profoundly linked to inflammatory bowel diseases (IBD), but little is known about the specific biological functions of miRNAs in IBD. This study sought to elucidate the effect and the underlying target of miR-29c-3p in ulcerative colitis (UC).
The levels of miR-29c-3p and leukemia inhibitory factor (LIF) were measured in inflamed lesions of UC patients and dextran sulfate sodium (DSS)-induced colitis mice by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. MiR-29c-3p was predicted to target LIF by bioinformatics software, which was verified via luciferase reporter assay and transfection of miR-29c-3p mimics or inhibitor. The role of miR-29c-3p/LIF axis in intestinal inflammation was explored in experimental colitis mice and Caco-2 cells.
MiR-29c-3p was markedly downregulated while LIF was upregulated in colon tissues of UC patients and DSS-challenged colitis mice as well as in primary intestinal epithelial cells (IECs) and LPS-treated Caco-2 cells. MiR-29c-3p inhibited LIF expression at the transcriptional level via binding to LIF 3'-untranslated region (UTR) in Caco-2 cells. Targeting miR-29c-3p/LIF axis regulated inflammatory cytokines production, cell proliferation and apoptosis. Overexpression of miR-29c-3p aggravated mice experimental colitis via suppressing LIF.
Our findings demonstrate that the upregulation of miR-29c-3p promotes gut inflammation and the expression of pro-inflammatory mediators via suppressing LIF, thereby modulating the pathogenesis of UC.
微小RNA（miRNA）失调与炎症性肠病（IBD）密切相关，但miRNA在IBD中的具体生物学功能尚不清楚。本研究旨在阐明miR-29c-3p在溃疡性结肠炎（UC）中的作用及潜在靶点。
通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测UC患者炎症病变组织及葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠中miR-29c-3p和白血病抑制因子（LIF）的水平。利用生物信息学软件预测miR-29c-3p靶向LIF，并通过荧光素酶报告基因检测及转染miR-29c-3p模拟物或抑制剂进行验证。在实验性结肠炎小鼠和Caco-2细胞中探讨miR-29c-3p/LIF轴在肠道炎症中的作用。
在UC患者及DSS诱导的结肠炎小鼠的结肠组织中，以及在原代肠上皮细胞（IECs）和LPS处理的Caco-2细胞中，miR-29c-3p显著下调，而LIF上调。在Caco-2细胞中，miR-29c-3p通过与LIF 3'非翻译区（UTR）结合在转录水平抑制LIF表达。靶向miR-29c-3p/LIF轴可调节炎性细胞因子的产生、细胞增殖和凋亡。miR-29c-3p过表达通过抑制LIF加重小鼠实验性结肠炎。
我们的研究结果表明，miR-29c-3p上调通过抑制LIF促进肠道炎症和促炎介质的表达，从而调节UC的发病机制。
