Zhu Meng-Xia, Huang Lin-Hui, Zhu Yi-Ke, Cai Xing-Jun
Department of Respiratory and Critical Care Medicine, Hainan General Hospital, Haikou, Hainan Province, China.
Exp Lung Res. 2021 Apr-May;47(4):161-172. doi: 10.1080/01902148.2021.1876792. Epub 2021 Feb 16.
Asthma is a chronic inflammatory heterogeneous respiratory disease. Previous studies showed that the lncRNA NEAT1 (nuclear paraspeckle assembly transcript 1) might play an important role in the pathogenesis of asthma, but its potential mechanism in airway smooth muscle cell (ASMC) inflammation remains largely unknown and needs further investigation. We performed cellular immunofluorescence to identify the features of ASMCs and detected the expression levels of lncRNA NEAT1, miR-139, TNF-α, IL-6, IL-8 and IL-1β by quantitative real-time PCR (Q-PCR) and ELISA. Western blotting (WB) was used to measure the protein expression of the related genes, and bioinformatics as well as dual luciferase assays were used to validate the interaction between lncRNA NEAT1 and miR-139 and the interaction between miR-139 and the 3'-UTR of JAK3. The expression of lncRNA NEAT1 was increased in the ASMCs of asthma patients, but miR-139 was decreased. Overexpression of lncRNA NEAT1 promoted the expression of the inflammatory cytokines such as TNF-α, IL-6, IL-8 and IL-1β in ASMCs. LncRNA NEAT1 was able to target miR-139 to activate the JAK3/STAT5 signaling pathway and induced the expression of these inflammatory cytokines in ASMCs. Overexpression of miR-139 or suppression of the JAK3/STAT5 signaling pathway reversed the inflammatory effect of lncRNA NEAT1. LncRNA NEAT1 played a pivotal role in ASMC inflammation and exerted its function through the miR-139/JAK3/STAT5 signaling network.
哮喘是一种慢性炎症性异质性呼吸系统疾病。先前的研究表明，长链非编码RNA NEAT1（核旁斑组装转录本1）可能在哮喘的发病机制中起重要作用，但其在气道平滑肌细胞（ASMC）炎症中的潜在机制仍 largely未知，需要进一步研究。我们进行了细胞免疫荧光以鉴定ASMC的特征，并通过定量实时PCR（Q-PCR）和ELISA检测长链非编码RNA NEAT1、miR-139、TNF-α、IL-6、IL-8和IL-1β的表达水平。蛋白质印迹法（WB）用于测量相关基因的蛋白质表达，生物信息学以及双荧光素酶测定用于验证长链非编码RNA NEAT1与miR-139之间的相互作用以及miR-139与JAK3的3'-UTR之间的相互作用。哮喘患者ASMC中长链非编码RNA NEAT1的表达增加，但miR-139降低。长链非编码RNA NEAT1的过表达促进了ASMC中TNF-α、IL-6、IL-8和IL-1β等炎性细胞因子的表达。长链非编码RNA NEAT1能够靶向miR-139以激活JAK3/STAT5信号通路，并诱导ASMC中这些炎性细胞因子的表达。miR-139的过表达或JAK3/STAT5信号通路的抑制逆转了长链非编码RNA NEAT1的炎症作用。长链非编码RNA NEAT1在ASMC炎症中起关键作用，并通过miR-139/JAK3/STAT5信号网络发挥其功能。
