Emergency Center, Zhongnan Hospital of Wuhan University, 430071, Wuhan, China.
Hubei Clinical Research Center for Emergency and Resuscitation, Zhongnan Hospital of Wuhan University, 430071, Wuhan, China.
BMC Genomics. 2021 Sep 18;22(1):671. doi: 10.1186/s12864-021-07970-y.
Ventilator-induced diaphragm dysfunction (VIDD) is a common complication of life support by mechanical ventilation observed in critical patients in clinical practice and may predispose patients to severe complications such as ventilator-associated pneumonia or ventilator discontinuation failure. To date, the alterations in microRNA (miRNA) expression in the rat diaphragm in a VIDD model have not been elucidated. This study was designed to identify these alterations in expression.
Adult male Wistar rats received conventional controlled mechanical ventilation (CMV) or breathed spontaneously for 12 h. Then, their diaphragm tissues were collected for RNA extraction. The miRNA expression alterations in diaphragm tissue were investigated by high-throughput microRNA-sequencing (miRNA-seq). For targeted mRNA functional analysis, gene ontology (GO) analyses and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were subsequently conducted. qRT-PCR validation and luciferase reporter assays were performed. We successfully constructed a model of ventilator-induced diaphragm dysfunction and identified 38 significantly differentially expressed (DE) miRNAs, among which 22 miRNAs were upregulated and 16 were downregulated. GO analyses identified functional genes, and KEGG pathway analyses revealed the signaling pathways that were most highly correlated, which were the MAPK pathway, FoxO pathway and Autophagy-animal. Luciferase reporter assays showed that STAT3 was a direct target of both miR-92a-1-5p and miR-874-3p and that Trim63 was a direct target of miR-3571.
The current research supplied novel perspectives on miRNAs in the diaphragm, which may not only be implicated in diaphragm dysfunction pathogenesis but could also be considered as therapeutic targets in diaphragm dysfunction.
呼吸机诱导的膈肌功能障碍（VIDD）是临床实践中接受机械通气支持的危重症患者中常见的并发症，可能使患者易患呼吸机相关性肺炎或呼吸机撤离失败等严重并发症。迄今为止，尚未阐明 VIDD 模型中大鼠膈肌中 microRNA（miRNA）表达的改变。本研究旨在确定这些表达变化。
成年雄性 Wistar 大鼠接受常规控制机械通气（CMV）或自主呼吸 12 小时。然后，采集他们的膈肌组织进行 RNA 提取。通过高通量 microRNA 测序（miRNA-seq）研究膈肌组织中 miRNA 表达的改变。为了进行靶向 mRNA 功能分析，随后进行了基因本体论（GO）分析和京都基因与基因组百科全书（KEGG）通路分析。进行了 qRT-PCR 验证和荧光素酶报告基因检测。我们成功构建了呼吸机诱导的膈肌功能障碍模型，并鉴定出 38 个显著差异表达（DE）miRNA，其中 22 个 miRNA 上调，16 个 miRNA 下调。GO 分析鉴定出功能基因，KEGG 通路分析显示与信号通路最相关的是 MAPK 通路、FoxO 通路和 Autophagy-animal。荧光素酶报告基因检测表明 STAT3 是 miR-92a-1-5p 和 miR-874-3p 的直接靶标，Trim63 是 miR-3571 的直接靶标。
本研究为膈肌中的 miRNA 提供了新的视角，它们不仅可能与膈肌功能障碍的发病机制有关，而且可以作为膈肌功能障碍的治疗靶点。
