Microarray Analysis Reveals the Changes in Circular RNA Expression and Molecular Mechanisms in Mice With Ventilator-Induced Lung Injury

Abstract

Circular RNA (circRNA) expression profiles in lung tissues from mice with and without ventilator-induced lung injury (VILI) were analyzed using high-throughput sequencing and bioinformatics to clarify their potential role in VILI pathogenesis and provide valuable molecular markers for VILI diagnosis and treatment. A VILI mouse model was established using high-tidal volume ventilation, and lung tissue was stained with HE and TUNEL. The present study used high-throughput sequencing technology to analyze the expression profile of circRNAs in the lung tissue of mice with and without VILI. Bioinformatics was used to analyze the enrichment of differentially expressed circRNAs using Gene Ontology and KEGG to predict function. Among the top 10 circRNAs with significant differential expression, we used real-time quantitative polymerase chain reaction technology (qRT-PCR) to verify the accuracy of the high-throughput sequencing results and constructed the corresponding circRNA-miRNA-mRNA-specific binding network map using software prediction. The most upregulated circRNAs were novel_circ_0000899 and novel_circ_0014815, and the most downregulated circRNAs were novel_circ_0015069. A total of 14,347 circRNAs were detected using high-throughput sequencing. Compared to the control group, 285 circRNAs were abnormally and significantly expressed in the lung tissues of VILI mice (|log2(FC)| > 1, p < 0.05). A total of 171 circRNAs were significantly upregulated, and 114 circRNAs were significantly downregulated. Gene ontology analyses indicated that the differentially expressed circRNAs were involved in multiple biological functions, such as regulation of metabolic processes, protein phosphorylation, and chromatin organization. KEGG pathway analyses revealed that the Ras signaling pathway, rap1 signaling pathway, PI3K − Akt signaling pathway, and ECM receiver interaction were related to the differentially expressed circRNAs. The qRT-PCR verification results were generally consistent with the circRNA expression trends of the high-throughput sequencing data. The circRNA-miRNA-mRNA interaction network suggested that miRNAs and mRNAs related to circRNAs played a key role in VILI. Differentially expressed circRNAs were identified in the tissues of VILI mice using high-throughput sequencing combined with bioinformatics analysis, and the results lay a foundation for further study of the mechanism of circRNAs in the occurrence and development of VILI.

Rutin alleviates ventilator-induced lung injury by inhibiting NLRP3 inflammasome activation

“…Our previous studies showed that inflammation plays an important role in the occurrence and development of VILI. 24 , 25 , 26 , 27 The network pharmacology results showed that NLRP3 inflammasome activation played an important role in the rutin-VILI target network. We further detected whether rutin reduced NLRP3 inflammasome activation in the lung tissues of mice with VILI.…”

“…The mouse VILI model was established according to our previous studies. 24 , 25 Briefly, the experimental mice were anesthetized with 1% sodium pentobarbital (100 mg/kg), and a tracheotomy was performed. A 22G trocar (BD Biosciences, United States) was inserted, and the trachea was ligated to prevent air leakage.…”

Rutin targets AKT to inhibit ferroptosis in ventilator‐induced lung injury

“…The mice were randomized into the control group or the VILI group. The mice in the VILI group were anesthetized with 1% sodium pentobarbital (100 mg/kg), and continuous MV was applied for 4 h according to our previous study (S. Chen et al, 2022). The mice in the control group were intubated but not ventilated.…”

Functional roles of circular RNAs in lung injury

“…novel_circ_0000899 and novel_circ_0014815 were identified to be the most upregulated circRNAs, whereas novel_circ_0015069 was the most downregulated circRNA. These circRNAs were found to be involved in metabolic processes, and in the pathway of Ras, Rap1, and PI3K/Akt ( Chen et al, 2022 ).…”