Department of Internal Emergency Medicine and Critical Care, Shanghai East Hospital, Tong Ji University, Shanghai, China.
Medical School/Tongji University, Shanghai, China.
J Cell Mol Med. 2019 Oct;23(10):7111-7115. doi: 10.1111/jcmm.14577. Epub 2019 Aug 14.
Circular RNAs (circRNAs) make up a large class of non-coding RNAs and play important roles in the pathology of a variety of diseases. However, their roles in pulmonary macrophage polarization after sepsisinduced lung injury is unknown. In this study, mice were divided into two groups: Sham control group and cecal ligation and puncture (CLP)-induced ALI group. Macrophages were isolated from lung homogenates 24 hours after SCLP/CLP. We started with RNA-seq of circRNA changes in macrophages and validated by RT-PCR in the following experiments. A total of 4318 circRNAs were detected in the two groups. Of these, 11 and 126 circRNAs were found to be significantly upregulated and downregulated, respectively, compared to the control (p≤0.05, Fold Change ≥2). Differentially expressed circRNAs with a high foldchange (fold-change >4, P<0.05) were selected for validation by qRT-PCR, 10 of which were verified. Furthermore, the most differentially expressed circRNAs within all the comparisons were annotated in detail with circRNA/miRNA interaction information using miRNA target prediction software. The network of circRNA-miRNA-mRNA was illustrated by cytoscape software. Gene ontology analyses indicated the upregulated circRNAs were involved in the multiple biological functions such as regulation of mitochondrion distribution and Notch binding, while the down-regulated circRNAs mainly involved in the biological process as histone H3K27 methylation. KEGG pathway analysis revealed TGF-beta signaling pathway was related to the upregulated circRNAs. The present study provides a novel insight into the roles of circRNAs in pulmonary macrophage differentiation and polarization post septic lung injury.
环状 RNA（circRNAs）构成了一大类非编码 RNA，并在多种疾病的病理学中发挥重要作用。然而，它们在脓毒症诱导的肺损伤后肺巨噬细胞极化中的作用尚不清楚。在这项研究中，将小鼠分为两组：假手术对照（Sham control）组和盲肠结扎和穿孔（cecal ligation and puncture，CLP）诱导的急性肺损伤（ALI）组。在 CLP/盲肠结扎后 24 小时，从肺匀浆中分离巨噬细胞。我们首先对巨噬细胞中 circRNA 变化进行了 RNA-seq 分析，并在以下实验中通过 RT-PCR 进行了验证。在两组中检测到 4318 个 circRNAs。与对照组相比，其中 11 个和 126 个 circRNAs分别显著上调和下调（p≤0.05，倍数变化≥2）。选择差异表达倍数高（fold-change >4，P<0.05）的 circRNAs 通过 qRT-PCR 进行验证，其中 10 个得到验证。此外，使用 miRNA 靶标预测软件，对所有比较中差异表达最显著的 circRNAs 进行了详细注释，获得了 circRNA/miRNA 相互作用信息。通过 cytoscape 软件展示了 circRNA-miRNA-mRNA 的网络。GO 分析表明，上调的 circRNAs 参与了多个生物学功能，如调节线粒体分布和 Notch 结合，而下调的 circRNAs 主要参与了组蛋白 H3K27 甲基化等生物学过程。KEGG 通路分析表明 TGF-β 信号通路与上调的 circRNAs 相关。本研究为 circRNAs 在脓毒症诱导的肺损伤后肺巨噬细胞分化和极化中的作用提供了新的见解。
