Li Sen, Liang Shuling, Xie Shunyu, Chen Haixia, Huang Haoying, He Qixin, Zhang Huayan, Wang Xiaohui
Guangzhou Women and Children's Medical Center, State Key Laboratory of Respiratory Disease and Guangzhou Medical University, Guangzhou, Guangdong Province, People's Republic of China.
Guangdong Provincial Research Center for Child Health, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, Guangdong Province, People's Republic of China.
J Inflamm Res. 2024 Mar 5;17:1467-1480. doi: 10.2147/JIR.S448394. eCollection 2024.
Bronchopulmonary dysplasia (BPD) has become a major cause of morbidity and mortality in preterm infants worldwide, yet its pathogenesis and underlying mechanisms remain poorly understood. The present study sought to explore microRNA-mRNA regulatory networks and immune cells involvement in BPD through a combination of bioinformatic analysis and experimental validation.
MicroRNA and mRNA microarray datasets were obtained from the Gene Expression Omnibus (GEO) database. Differentially expressed microRNAs (DEMs) were identified in BPD patients compared to control subjects, and their target genes were predicted using miRWalk, miRNet, miRDB, and TargetScan databases. Subsequently, protein-protein interaction (PPI) and functional enrichment analyses were conducted on the target genes. 30 hub genes were screened using the Cytohubba plugin of the Cytoscape software. Additionally, mRNA microarray data was utilized to validate the expression of hub genes and to perform immune infiltration analysis. Finally, real-time PCR (RT-PCR), immunohistochemistry (IHC), and flow cytometry were conducted using a mouse model of BPD to confirm the bioinformatics findings.
Two DEMs (miR-15b-5p and miR-20a-5p) targeting genes primarily involved in the regulation of cell cycle phase transition, ubiquitin ligase complex, protein serine/threonine kinase activity, and MAPK signaling pathway were identified. and four autophagy-related genes (, and ) were differentially expressed in the mRNA microarray dataset. Analysis of immune infiltration revealed significant differences in levels of neutrophils and naive B cells between BPD patients and control subjects. RT-PCR and IHC confirmed reduced expression of in a mouse model of BPD. Although the proportion of total neutrophils did not change appreciably, the activation of neutrophils, marked by loss of CD62L, was significantly increased in BPD mice.
Downregulation of mediated by miR-15b-5p and miR-20a-5p may be associated with the development of BPD. Additionally, increased CD62L neutrophil subset might be important for the immune-mediated injury in BPD.
支气管肺发育不良（BPD）已成为全球早产儿发病和死亡的主要原因，但其发病机制和潜在机制仍知之甚少。本研究旨在通过生物信息学分析和实验验证相结合的方法，探索微小RNA-信使核糖核酸调控网络以及免疫细胞在BPD中的作用。
从基因表达综合数据库（GEO）中获取微小RNA和信使核糖核酸微阵列数据集。与对照受试者相比，在BPD患者中鉴定出差异表达的微小RNA（DEM），并使用miRWalk、miRNet、miRDB和TargetScan数据库预测其靶基因。随后，对靶基因进行蛋白质-蛋白质相互作用（PPI）和功能富集分析。使用Cytoscape软件的Cytohubba插件筛选出30个枢纽基因。此外，利用信使核糖核酸微阵列数据验证枢纽基因的表达并进行免疫浸润分析。最后，使用BPD小鼠模型进行实时聚合酶链反应（RT-PCR）、免疫组织化学（IHC）和流式细胞术，以证实生物信息学研究结果。
鉴定出两个主要参与细胞周期相变调控、泛素连接酶复合物、蛋白质丝氨酸/苏氨酸激酶活性和丝裂原活化蛋白激酶（MAPK）信号通路调控的靶基因的DEM（miR-15b-5p和miR-20a-5p）。并且四个自噬相关基因（……此处原文缺失具体基因名称……）在信使核糖核酸微阵列数据集中差异表达。免疫浸润分析显示，BPD患者和对照受试者之间中性粒细胞和幼稚B细胞水平存在显著差异。RT-PCR和IHC证实BPD小鼠模型中……此处原文缺失具体基因名称……的表达降低。尽管中性粒细胞总数比例没有明显变化，但以CD62L缺失为标志的中性粒细胞活化在BPD小鼠中显著增加。
miR-15b-5p和miR-20a-5p介导的……此处原文缺失具体基因名称……下调可能与BPD的发生发展有关。此外，CD62L中性粒细胞亚群增加可能对BPD中的免疫介导损伤很重要。
