Department of Pulmonary and Critical Care Medicine, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Shandong Key Laboratory of Infectious Respiratory Diseases, China.
Department of Pulmonary Disease, Jinan Traditional Chinese Medicine Hospital, China.
Biomed Res Int. 2021 Oct 21;2021:6831770. doi: 10.1155/2021/6831770. eCollection 2021.
Acute lung injury (ALI) is a fatal syndrome frequently induced by lipopolysaccharide (LPS) released from the bacterial cell wall. LPS could also trigger autophagy of lung bronchial epithelial cell to relieve the inflammation, while the overwhelming LPS would impair the balance of autophagy consequently inducing serious lung injury.
We observed the autophagy variation of 16HBE, human bronchial epithelial cell, under exposure to different concentrations of LPS through western blot, immunofluorescence staining, and electron microscopy. Eight strands of 16HBE were divided into two groups upon 1000 ng/ml LPS stimulation or not, which were sent to be sequenced at whole transcriptome. Subsequently, we analyzed the sequencing data in functional enrichment, pathway analysis, and candidate gene selection and constructed a hsa-miR-663b-related competing endogenous RNA (ceRNA) network.
We set a series of concentrations of LPS to stimulate 16HBE and observed the variation of autophagy in related protein expression and autophagosome count. We found that the effective concentration of LPS was 1000 ng/ml at 12 hours of exposure and sequenced the 1000 ng/ml LPS-stimulated 16HBE. As a result, a total of 750 differentially expressed genes (DEGs), 449 differentially expressed lncRNAs (DElncRNAs), 76 differentially expressed circRNAs (DEcircRNAs), and 127 differentially expressed miRNAs (DEmiRNAs) were identified. We constructed the protein-protein interaction (PPI) network to visualize the interaction between DEGs and located 36 genes to comprehend the core discrepancy between LPS-stimulated 16HBE and the negative control group. In combined analysis of differentially expressed RNAs (DERNAs), we analyzed all the targeted relationships of ceRNA in DERNAs and figured hsa-miR-663b as a central mediator in the ceRNA network to play when LPS induced the variation of autophagy in 16HBE.
Our research indicated that the hsa-miR-663b-related ceRNA network may contribute to the key regulatory mechanism in LPS-induced changes of autophagy and ALI.
急性肺损伤（ALI）是一种致命的综合征，常由细菌细胞壁释放的脂多糖（LPS）引起。LPS 也可以触发肺支气管上皮细胞的自噬来缓解炎症，而过量的 LPS 会破坏自噬的平衡，从而导致严重的肺损伤。
我们通过 Western blot、免疫荧光染色和电子显微镜观察不同浓度 LPS 暴露下 16HBE（人支气管上皮细胞）自噬的变化。将 16HBE 的 8 条链分为 LPS 刺激组和非刺激组，每组各 4 条链，用全转录组测序。随后，我们对测序数据进行功能富集、通路分析和候选基因选择，并构建了 hsa-miR-663b 相关竞争内源性 RNA（ceRNA）网络。
我们用一系列浓度的 LPS 刺激 16HBE，观察相关蛋白表达和自噬体数量的变化。我们发现 LPS 的有效浓度为 1000ng/ml，在暴露 12 小时后进行测序。结果共鉴定出 750 个差异表达基因（DEGs）、449 个差异表达长链非编码 RNA（DElncRNA）、76 个差异表达环状 RNA（DEcircRNA）和 127 个差异表达 miRNA（DEmiRNA）。我们构建了蛋白质-蛋白质相互作用（PPI）网络，以可视化 DEGs 之间的相互作用，并找到了 36 个基因，以理解 LPS 刺激的 16HBE 与阴性对照组之间的核心差异。在差异表达 RNA（DERNA）的综合分析中，我们分析了 DERNAs 中 ceRNA 的所有靶向关系，并发现 hsa-miR-663b 是 ceRNA 网络中的一个中心调节因子，在 LPS 诱导 16HBE 自噬变化时发挥作用。
我们的研究表明，hsa-miR-663b 相关的 ceRNA 网络可能有助于 LPS 诱导的自噬和 ALI 变化的关键调节机制。
