MicroRNA-485-3p通过激活人气道平滑肌细胞Wnt/β-Catenin信号通路促进炎症反应和细胞外基质沉积

IF 1.5 4区 医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Critical Reviews in Eukaryotic Gene Expression Pub Date : 2023-01-01 DOI:10.1615/CritRevEukaryotGeneExpr.2022044300
Cuiyun Liu, Sen Shi, Ying Gao, Qian Leng, Rui Gong, Lan Zhang, Jinhai Ma
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引用次数: 0

摘要

本研究旨在研究microRNA (miR)-485-3p对人气道平滑肌细胞(HASMCs)炎症反应和细胞外基质沉积的影响。采用实时定量聚合酶链式反应(qRT-PCR)检测小儿哮喘(PA)患者和对照组外周血中miR-485-3p和WIF1的水平。将miR-485-3p抑制剂和模拟物以及阴性对照(NC)抑制剂/模拟物转染到经肿瘤坏死因子(TNF)-α处理的HASMCs中。采用酶联免疫吸附试验(ELISA)分析eotaxin、白细胞介素(IL)-8、IL-6水平。同时进行纤维连接蛋白的细胞免疫荧光分析。采用TargetScan和双荧光素酶报告基因法预测和验证miR-485-3p的靶基因。Western blot检测IL-6、eotaxin、IL-8、collagen III、collagen I、MMP-9、TIMP-1、MMP-2、axin、β-catenin、磷酸化β-catenin、GSK3β、p-GSK3β、WIF1蛋白水平。PA患者miR-485-3p水平升高,而WIF1表达较低。在TNF-α-诱导的HASMCs中,miR-485-3p过表达促进了炎症反应和细胞外基质的积累。WIF1是miR-485-3p的直接靶点。沉默miR-485-3p可抑制Wnt/β-catenin信号通路的激活。沉默miR-485-3p引起的炎症反应和ECM积累的减少是通过阻断Wnt/β-catenin信号传导诱导的。因此,miRNA-485-3p靶向WIF1并激活Wnt/β-catenin信号,促进炎症反应的激活和HASMCs中ECM的积累。
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MicroRNA-485-3p Promotes the Inflammatory Response and Extracellular Matrix Deposition by Activating Wnt/β-Catenin Signaling in Human Airway Smooth Muscle Cells.

The aim of this study was to study the effects of microRNA (miR)-485-3p on the inflammatory response and extracellular matrix deposition of human airway smooth muscle cells (HASMCs). The levels of miR-485-3p and WIF1 in peripheral blood of pediatric asthma (PA) patients and controls were examined by quantitative real-time polymerase chain reaction (qRT-PCR). miR-485-3p inhibitor and mimic, together with negative control (NC) inhibitor/ mimic, were transfected into HASMCs treated with tumor necrosis factor (TNF)-α. The levels of eotaxin, interleukin (IL)-8, and IL-6 were analyzed by enzyme-linked immunosorbent assay (ELISA). Cellular immunofluorescence analysis of fibronectin was also performed. The target genes of miR-485-3p were predicted and validated using TargetScan and dual-luciferase reporter gene assay. The protein levels of IL-6, eotaxin, IL-8, collagen III, collagen I, MMP-9, TIMP-1, MMP-2, axin, β-catenin, phosphorylated β-catenin, GSK3β, p-GSK3β, and WIF1 were tested by Western blot. The level of miR-485-3p was increased, whereas expression of WIF1 was low in PA patients. In TNF-α-induced HASMCs, miR-485-3p overexpression promoted the inflammatory response and the accumulation of extracellular matrix. WIF1 was a direct target of miR-485-3p. Silencing miR-485-3p inhibited activation of Wnt/β-catenin signaling. The reductions in the inflammatory response and ECM accumulation caused by silencing miR-485-3p were induced by blocking Wnt/β-catenin signaling. Thus, miRNA-485-3p targets WIF1 and activates Wnt/β-catenin signaling, facilitating activation of the inflammatory response and ECM accumulation in HASMCs.

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来源期刊
Critical Reviews in Eukaryotic Gene Expression
Critical Reviews in Eukaryotic Gene Expression 生物-生物工程与应用微生物
CiteScore
2.70
自引率
0.00%
发文量
67
审稿时长
1 months
期刊介绍: Critical ReviewsTM in Eukaryotic Gene Expression presents timely concepts and experimental approaches that are contributing to rapid advances in our mechanistic understanding of gene regulation, organization, and structure within the contexts of biological control and the diagnosis/treatment of disease. The journal provides in-depth critical reviews, on well-defined topics of immediate interest, written by recognized specialists in the field. Extensive literature citations provide a comprehensive information resource. Reviews are developed from an historical perspective and suggest directions that can be anticipated. Strengths as well as limitations of methodologies and experimental strategies are considered.
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