Nowakowska Joanna, Kachel Maria, Langwiński Wojciech, Ziarniak Kamil, Szczepankiewicz Aleksandra
Molecular and Cell Biology Unit, Department of Pediatric Pulmonology, Allergy and Clinical Immunology, Poznan University of Medical Sciences, 60-572 Poznan, Poland.
Doctoral School, Poznan University of Medical Sciences, 60-812 Poznan, Poland.
Cells. 2025 Jan 12;14(2):104. doi: 10.3390/cells14020104.
Asthma is a major non-communicable disease whose pathogenesis is still not fully elucidated. One of the asthma research models is precision-cut lung slices (PCLSs), and among the therapeutic options, miRNA molecules are of great interest. The aim of our study was to investigate whether inhibition of miR-223-3p and miR328a-3p affects the inflammatory response in PCLSs derived from a rat with HDM-induced allergic inflammation and a control rat. We generated rat PCLSs and transfected them with miR-223-3p and miR-328a-3p inhibitors. RNA was isolated from PCLSs and analyzed by qPCR. We also examined the proteins in the culture medium using the Magnetic Luminex Assay. The comparison between miRNA-transfected PCLSs and non-transfected controls showed significant differences in the expression of several genes associated with allergic inflammation, including , , and , in both the rat with allergic inflammation and the control rat. In the culture medium, we found no significant differences in protein levels between rat with allergic inflammation and the control. Our study highlighted some important issues: the need to extend the model by including more biological replicates, the need to standardize culture conditions, and the need to consider co-transfection with several miRNA inhibitors when modifying miRNAs expression in the PCLS model.
哮喘是一种主要的非传染性疾病，其发病机制仍未完全阐明。哮喘研究模型之一是精密肺切片（PCLSs），在治疗选择中，miRNA分子备受关注。我们研究的目的是调查抑制miR-223-3p和miR328a-3p是否会影响来自经屋尘螨诱导的过敏性炎症大鼠和对照大鼠的PCLSs中的炎症反应。我们制备了大鼠PCLSs，并用miR-223-3p和miR-328a-3p抑制剂转染它们。从PCLSs中分离RNA并通过qPCR进行分析。我们还使用磁珠Luminex检测法检测了培养基中的蛋白质。miRNA转染的PCLSs与未转染的对照之间的比较显示，在过敏性炎症大鼠和对照大鼠中，与过敏性炎症相关的几个基因（包括 、 、 和 ）的表达存在显著差异。在培养基中，我们发现过敏性炎症大鼠和对照之间的蛋白质水平没有显著差异。我们的研究突出了一些重要问题：需要通过纳入更多生物学重复来扩展模型，需要标准化培养条件，以及在PCLS模型中改变miRNA表达时需要考虑与几种miRNA抑制剂共转染。
