circ_0026579 通过靶向 miR-338-3p/TBL1XR1 轴减轻脂多糖（细菌来源）诱导的支气管上皮细胞炎症损伤。
Knockdown of circ_0026579 ameliorates lipopolysaccharide (bacterial origin)-induced inflammatory injury in bronchial epithelium cells by targeting miR-338-3p/TBL1XR1 axis.
机构信息
Department of Paediatrics, First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine, Hangzhou, Zhejiang, China.
Department of Paediatrics, First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine, Hangzhou, Zhejiang, China.
出版信息
Transpl Immunol. 2022 Oct;74:101635. doi: 10.1016/j.trim.2022.101635. Epub 2022 May 27.
BACKGROUND
Circular RNAs (circRNAs) play an important regulatory role in human diseases including organ allograft rejection. The aim of this study is to clarify the functional role and molecular mechanism of circ_0026579 RNA in lipopolysaccharide (LPS)-induced bronchopneumonia injury.
MATERIALS AND METHODS
Bronchial epithelial BEAS-2B cells were treated with LPS to mimic an in vitro model for bronchopneumonia. Cell viability and proliferation were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 5-ethynyl-2'-deoxyuridine (EdU) assay. Flow cytometry assay was used to assess cell apoptosis. Caspase-3 activity was analyzed by Caspase-3 activity assay kit. The expression levels of circ_0026579 RNA, miR-338-3p, and transducin β-like 1× related protein 1 (TBL1XR1) RNA were determined by RT-qPCR. The protein level was quantified by western blot assay. The correlation between miR-338-3p and circ_0026579 or TBL1XR1 was confirmed by dual-luciferase reporter and RNA immunoprecipitation assays.
RESULTS
LPS treatment repressed proliferation but induced apoptosis and inflammatory response in BEAS-2B cells. Circ_0026579 RNA was highly expressed in patients with pneumonia. Besides, the expression levels of circ_0026579 RNA and TBL1XR1 RNA/protein were upregulated, while miR-338-3p level was decreased in LPS-treated BEAS-2B cells. Knockdown of circ_0026579 RNA or TBL1XR1 protein could abolish LPS-induced cell injury in BEAS-2B cells. Furthermore, we found that circ_0026579 RNA functioned as a "sponge" for miR-338-3p to regulate TBL1XR1 expression. Additionally, silencing circ_0026579 RNA protected BEAS-2B cells from LPS-induced bronchopneumonia injury by regulating TBL1XR1 expression.
CONCLUSION
Circ_0026579 RNA knockdown promoted cell proliferation but inhibited apoptosis and inflammation in LPS-induced BEAS-2B cells through regulating miR-338-3p RNA/TBL1XR1 protein axis.
背景
环状 RNA（circRNAs）在包括器官移植排斥在内的人类疾病中发挥着重要的调控作用。本研究旨在阐明 circ_0026579 RNA 在脂多糖（LPS）诱导的支气管肺炎损伤中的功能作用和分子机制。
材料和方法
用 LPS 处理支气管上皮 BEAS-2B 细胞，模拟支气管肺炎的体外模型。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测和 5-乙炔基-2'-脱氧尿苷（EdU）检测分析细胞活力和增殖。流式细胞术检测细胞凋亡。通过 Caspase-3 活性检测试剂盒分析 Caspase-3 活性。通过 RT-qPCR 检测 circ_0026579 RNA、miR-338-3p 和转导素β样 1×相关蛋白 1（TBL1XR1）RNA 的表达水平。通过 Western blot 检测蛋白水平。通过双荧光素酶报告和 RNA 免疫沉淀检测证实 miR-338-3p 与 circ_0026579 或 TBL1XR1 的相关性。
结果
LPS 处理抑制了 BEAS-2B 细胞的增殖，但诱导了细胞凋亡和炎症反应。肺炎患者中 circ_0026579 RNA 表达水平升高。此外，LPS 处理的 BEAS-2B 细胞中 circ_0026579 RNA 和 TBL1XR1 RNA/蛋白表达水平上调，而 miR-338-3p 水平下调。circ_0026579 RNA 或 TBL1XR1 蛋白的敲低可消除 LPS 诱导的 BEAS-2B 细胞损伤。此外，我们发现 circ_0026579 RNA 作为 miR-338-3p 的“海绵”发挥作用，调节 TBL1XR1 的表达。此外，沉默 circ_0026579 RNA 通过调节 TBL1XR1 表达来保护 BEAS-2B 细胞免受 LPS 诱导的支气管肺炎损伤。
结论
circ_0026579 RNA 敲低通过调节 miR-338-3p RNA/TBL1XR1 蛋白轴促进 LPS 诱导的 BEAS-2B 细胞增殖，但抑制细胞凋亡和炎症。
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