Department of Immunology, IIS-Fundación Jiménez Díaz, Madrid, Spain.
CIBER de Enfermedades Respiratorias (CIBERES), Madrid, Spain.
J Cell Physiol. 2022 Feb;237(2):1498-1508. doi: 10.1002/jcp.30620. Epub 2021 Oct 26.
Asthma is a chronic respiratory disease produced by an aberrant immune response that originates with breathing difficulties and cough, through airway remodeling. The above pathophysiological events of asthma emerge the regulators of effectors, like epigenetics, which include microRNAs (miRNAs) who perform post-transcriptional regulation, controlling diverse pathways in respiratory diseases. The objective of the study was to determine how miR-185-5p regulates the secretion of periostin by airway structural cells, and smooth muscle cells contraction, both related to airway remodeling in asthma. We used miR-185-5p mimic and inhibitors in bronchial smooth muscle cells (BSMCs) and small airway epithelial cells (SAECs) from healthy subjects. Gene expression and protein levels of periostin (POSTN), CDC42, and RHOA were analyzed by RT-PCR and ELISA/Western blot, respectively. BSMC contractility was analyzed using cell-embedded collagen gels and measurement of intracellular calcium was performed using Fura-2. Additionally, miR-185-5p and periostin expression were evaluated in sputum from healthy and asthmatics. From these experiments, we observed that miR-185-5p modulation regulates periostin mRNA and protein in BSMCs and SAECs. A tendency for diminished miR-185-5p expression and higher periostin levels was seen in sputum cells from asthmatics compared to healthy, with an inverse correlation observed between POSTN and miR-185-5p. Inhibition of miR-185-5p produced higher BSMCs contraction induced by histamine. Calcium mobilization was not modified by miR-185-5p, showing that miR-185-5p role in BSMC contractility is performed by regulating CDC42 and RhoA pro-contractile factors instead. In conclusion, miR-185-5p is a modulator of periostin secretion by airway structural cells and of smooth muscle contraction, which can be related to asthma pathophysiology, and thus, might be a promising therapeutic target.
哮喘是一种慢性呼吸道疾病,由异常免疫反应引起,最初表现为呼吸困难和咳嗽,并伴有气道重塑。哮喘的上述病理生理事件出现了效应器的调节剂,如表观遗传学,包括 microRNAs (miRNAs),它们进行转录后调节,控制呼吸道疾病中的多种途径。本研究的目的是确定 miR-185-5p 如何调节气道结构细胞periostin 的分泌和平滑肌细胞收缩,这两者都与哮喘中的气道重塑有关。我们使用 miR-185-5p 模拟物和抑制剂在来自健康受试者的支气管平滑肌细胞 (BSMCs) 和小气道上皮细胞 (SAECs) 中进行实验。通过 RT-PCR 和 ELISA/Western blot 分别分析 periostin (POSTN)、CDC42 和 RHOA 的基因表达和蛋白水平。使用细胞嵌入胶原凝胶分析 BSMC 收缩性,并使用 Fura-2 测量细胞内钙。此外,还评估了健康人和哮喘患者痰液中的 miR-185-5p 和 periostin 表达。通过这些实验,我们观察到 miR-185-5p 调节可调节 BSMCs 和 SAECs 中的 periostin mRNA 和蛋白。与健康人相比,哮喘患者痰液细胞中的 miR-185-5p 表达下调和 periostin 水平升高的趋势更为明显,POSTN 与 miR-185-5p 之间存在负相关关系。miR-185-5p 抑制可导致组胺诱导的 BSMC 收缩增加。miR-185-5p 不改变钙动员,表明 miR-185-5p 在 BSMC 收缩性中的作用是通过调节 CDC42 和 RhoA 促收缩因子来实现的。总之,miR-185-5p 是气道结构细胞 periostin 分泌和平滑肌收缩的调节剂,可能与哮喘病理生理学有关,因此可能是一种有前途的治疗靶点。