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哮喘患儿中微小RNA-378的表达升高,通过促进气道平滑肌细胞的增殖和抗凋亡作用加重气道重塑。

Elevated expression of microRNA-378 in children with asthma aggravates airway remodeling by promoting the proliferation and apoptosis resistance of airway smooth muscle cells.

作者信息

Li Peng, Lang Xufang, Xia Shungang

机构信息

Department of Pediatrics, Maternity and Child Health Care Hospital of Zibo City, Zibo, Shandong 255029, P.R. China.

Department of Student Affairs, College of Nursing, Zibo Vocational Institute, Zibo, Shandong 255314, P.R. China.

出版信息

Exp Ther Med. 2019 Mar;17(3):1529-1536. doi: 10.3892/etm.2018.7141. Epub 2018 Dec 28.

Abstract

The present study determined the expression of microRNA (miR)-378 in the peripheral blood and lung tissues of children with asthma, and investigated its effect and mechanism of action on the biological functions of airway smooth muscle cells. A total of 23 asthmatic children and 15 healthy children were included in the study. Peripheral blood and tissues were obtained from asthmatic children. Healthy children provided peripheral blood. Quantitative real-time polymerase chain reaction was used to determine the expression of miR-378. Airway smooth muscle cells were isolated and cultured . The cells were transfected with miR-378 mimics or miR-378 inhibitor. Following transfection, proliferation of the cells was determined using the CCK-8 assay. In addition, flow cytometry was used to detect the cell cycles and apoptosis of smooth muscle cells. Western blotting was performed to determine the expression of extracellular matrix proteins in smooth muscle cells. Furthermore, bioinformatics was used to predict potential target genes of miR-378 and their downstream signaling pathways. Results indicated that the expression of miR-378 in peripheral blood and lung tissues from asthmatic children was increased compared with that in healthy children. Serum from asthmatic children promoted the proliferation of smooth muscle cells by affecting the cell cycle, and enhanced apoptotic resistance of smooth muscle cells. Notably, overexpression of miR-378 increased the proliferation of smooth muscle cells by affecting the cell cycle, and this upregulated apoptotic resistance of smooth muscle cells and enhanced the expression of extracellular matrix-related proteins in smooth muscle cells. However, downregulation of miR-378 expression reversed the promoting effect of serum from asthmatic children on the biological functions of smooth muscle cells. These findings suggested that miR-378 possibly affects the proliferation, apoptosis and motility of airway smooth muscle cells via downstream signaling pathways. To conclude, the present study demonstrated that miR-378 expression was elevated in the peripheral blood and lung tissues from children with asthma. Furthermore, miR-378 promoted the biological functions of extracellular matrix-related proteins of smooth muscle cells, and possibly exerts its effect via its target genes through downstream signaling pathways.

摘要

本研究测定了哮喘患儿外周血和肺组织中微小RNA(miR)-378的表达,并研究了其对气道平滑肌细胞生物学功能的影响及作用机制。本研究共纳入23例哮喘患儿和15例健康儿童。采集哮喘患儿的外周血和组织。健康儿童提供外周血。采用定量实时聚合酶链反应测定miR-378的表达。分离并培养气道平滑肌细胞。用miR-378模拟物或miR-378抑制剂转染细胞。转染后,采用CCK-8法测定细胞增殖情况。此外,采用流式细胞术检测平滑肌细胞的细胞周期和凋亡情况。进行蛋白质印迹法以测定平滑肌细胞中细胞外基质蛋白的表达。此外,利用生物信息学预测miR-378的潜在靶基因及其下游信号通路。结果表明,与健康儿童相比,哮喘患儿外周血和肺组织中miR-378的表达升高。哮喘患儿的血清通过影响细胞周期促进平滑肌细胞增殖,并增强平滑肌细胞的抗凋亡能力。值得注意的是,miR-378的过表达通过影响细胞周期增加平滑肌细胞的增殖,上调平滑肌细胞的抗凋亡能力并增强平滑肌细胞中细胞外基质相关蛋白的表达。然而,miR-378表达的下调逆转了哮喘患儿血清对平滑肌细胞生物学功能的促进作用。这些发现提示,miR-378可能通过下游信号通路影响气道平滑肌细胞的增殖、凋亡和迁移。总之,本研究表明,哮喘患儿外周血和肺组织中miR-378的表达升高。此外,miR-378促进平滑肌细胞中细胞外基质相关蛋白的生物学功能,并可能通过其靶基因经下游信号通路发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6784/6364182/fef100ffe709/etm-17-03-1529-g00.jpg

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