Department of Intensive Care Unit, Key Laboratory for Critical Care Medicine of the Ministry of Health, Emergency Medicine Research Institute, Tianjin First Center Hospital, Nankai University, Tianjin, China.
State Key Laboratory of Medicinal Chemical Biology, Research Center for Analytical Sciences, Tianjin Key Laboratory of Biosensing and Molecular Recognition, College of Chemistry, Nankai University, Tianjin, China.
Exp Lung Res. 2024;50(1):242-258. doi: 10.1080/01902148.2024.2429184. Epub 2024 Nov 25.
In this study, we identified differentially expressed genes (DEGs) and signaling pathways to gain insight into the pathogenesis of acute lung injury (ALI). C57BL/6 mice were intravenously injected with lipopolysaccharide (LPS) to establish a sepsis-induced ALI model. Hematoxylin-eosin (H&E) and enzyme-linked immunosorbent assays (ELISAs) were used to evaluate the model. Whole transcriptome sequencing was performed to identify the expression changes in lncRNAs, circRNAs, miRNAs and mRNAs in lung tissues. The crucial RNAs and the biological function of the target genes were confirmed and annotated based on bioinformatics analysis. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was employed to verify the expression levels of key lncRNAs, circRNAs, miRNAs and mRNAs in the lung tissues and human bronchoalveolar lavage (BALF). A total of 3304 (1632 upregulated and 1672 downregulated) differentially expressed mRNAs, 794 (397 up and 397 down) differentially expressed lncRNAs, 89 (58 up and 31 down) differentially expressed circRNAs, and 14 (11 up and 3 down) differentially expressed miRNAs were identified between the control and LPS lung tissues. The lncRNA ceRNA subnetwork and circRNA ceRNA subnetwork were constructed based on the observed interaction and co-expression among the differentially expressed RNAs. An analysis of the protein-protein interaction (PPI) network and hub genes revealed crucial mRNAs for circRNA-Tcf20. The lncRNA-Snhg12, miR-212-3p and miR-223-3p were upregulated in sepsis ARDS patients. CircRNA-Tcf20, were significantly downregulated in sepsis ARDS patients. The biological function analysis indicated that these genes were enriched in the TNF signaling pathway, Necroptosis signaling pathway and the PI3K-Akt signaling pathway. Our findings suggest that circRNA-Tcf20, miR-212-3p, miR-223-3p, may be new regulatory factors that participate in the pathogenesis of sepsis-related acute lung injury. CircRNA-Tcf20, lncRNA-Snhg12 and all the other RNAs may be potential biomarkers for septic ALI/ARDS.
在这项研究中，我们鉴定了差异表达基因（DEGs）和信号通路，以深入了解急性肺损伤（ALI）的发病机制。通过静脉注射脂多糖（LPS）建立 C57BL/6 小鼠脓毒症诱导的 ALI 模型。苏木精-伊红（H&E）和酶联免疫吸附测定（ELISA）用于评估模型。进行全转录组测序以鉴定肺组织中 lncRNA、circRNA、miRNA 和 mRNA 的表达变化。基于生物信息学分析，鉴定并注释关键 RNA 及其靶基因的生物学功能。实时定量逆转录聚合酶链反应（qRT-PCR）用于验证肺组织和人支气管肺泡灌洗液（BALF）中关键 lncRNA、circRNA、miRNA 和 mRNA 的表达水平。在对照和 LPS 肺组织之间共鉴定出 3304 个（1632 个上调和 1672 个下调）差异表达的 mRNA、794 个（397 个上调和 397 个下调）差异表达的 lncRNA、89 个（58 个上调和 31 个下调）差异表达的 circRNA 和 14 个（11 个上调和 3 个下调）差异表达的 miRNA。基于观察到的差异表达 RNA 之间的相互作用和共表达，构建了 lncRNA ceRNA 子网络和 circRNA ceRNA 子网络。分析蛋白质-蛋白质相互作用（PPI）网络和关键基因揭示了 circRNA-Tcf20 的关键 mRNAs。lncRNA-Snhg12、miR-212-3p 和 miR-223-3p 在脓毒症 ARDS 患者中上调。CircRNA-Tcf20 在脓毒症 ARDS 患者中显著下调。生物学功能分析表明，这些基因富集在 TNF 信号通路、坏死性凋亡信号通路和 PI3K-Akt 信号通路中。我们的研究结果表明，circRNA-Tcf20、miR-212-3p、miR-223-3p 可能是参与脓毒症相关急性肺损伤发病机制的新调节因子。CircRNA-Tcf20、lncRNA-Snhg12 和所有其他 RNA 可能是脓毒症性急性肺损伤/急性呼吸窘迫综合征的潜在生物标志物。
