PDF(Pubmed)
Abstract：
Circular RNAs have emerged as important regulators in the pathogenesis of human diseases, including infantile pneumonia (IP). In this study, we aimed to explore the effects of circ_0035292 on lipopolysaccharide (LPS)-treated Wistsar Institute (WI)-38 cells.
Quantitative real-time polymerase chain reaction and western blot were executed to detect the levels of circ_0035292, microRNA-370-3p (miR-370-3p) and transducin β-like 1X related protein 1 (TBL1XR1). Cell counting kit-8, 5-ethynyl-2\'-deoxyuridine, and flow cytometry assessed cell proliferation and apoptosis. Concentrations of inflammatory factors were examined with enzyme linked immunosorbent assay kits. Dual-luciferase reporter assay and RNA immunoprecipitation were adopted to analyze binding between miR-370-3p and circ_0035292 or TBL1XR1.
Circ_0035292 level was increased in IP patients and LPS-triggered WI-38 cells. Circ_0035292 knockdown rescued LPS-mediated WI-38 cell proliferation suppression and WI-38 cell apoptosis and inflammation promotion. Circ_0035292 interacted with miR-370-3p and miR-370-3p directly targeted TBL1XR1. Moreover, miR-370-3p overexpression alleviated LPS-induced WI-38 cell apoptosis and inflammatory injury, which was abrogated via TBL1XR1 upregulation. Circ_0035292 absence inhibited the NF-κB pathway.
Knockdown of circ_0035292 rescued LPS-triggered WI-38 cell injury via miR-370-3p/TBL1XR1 axis and NF-κB pathway.
Quantitative real-time polymerase chain reaction and western blot were executed to detect the levels of circ_0035292, microRNA-370-3p (miR-370-3p) and transducin β-like 1X related protein 1 (TBL1XR1). Cell counting kit-8, 5-ethynyl-2\'-deoxyuridine, and flow cytometry assessed cell proliferation and apoptosis. Concentrations of inflammatory factors were examined with enzyme linked immunosorbent assay kits. Dual-luciferase reporter assay and RNA immunoprecipitation were adopted to analyze binding between miR-370-3p and circ_0035292 or TBL1XR1.
Circ_0035292 level was increased in IP patients and LPS-triggered WI-38 cells. Circ_0035292 knockdown rescued LPS-mediated WI-38 cell proliferation suppression and WI-38 cell apoptosis and inflammation promotion. Circ_0035292 interacted with miR-370-3p and miR-370-3p directly targeted TBL1XR1. Moreover, miR-370-3p overexpression alleviated LPS-induced WI-38 cell apoptosis and inflammatory injury, which was abrogated via TBL1XR1 upregulation. Circ_0035292 absence inhibited the NF-κB pathway.
Knockdown of circ_0035292 rescued LPS-triggered WI-38 cell injury via miR-370-3p/TBL1XR1 axis and NF-κB pathway.
摘要：
背景：环状RNA已成为人类疾病发病机制中的重要调节因子，包括小儿肺炎（IP）。在这项研究中,我们旨在探讨circ_0035292对脂多糖（LPS）处理的Wistsar研究所（WI）-38细胞的影响。
方法：进行定量实时聚合酶链反应和蛋白质印迹检测circ_0035292，microRNA-370-3p（miR-370-3p）和转导蛋白β样1X相关蛋白1（TBL1XR1）的水平。细胞计数试剂盒-8,5-乙炔基-2'-脱氧尿苷,流式细胞术评估细胞增殖和凋亡。用酶联免疫吸附测定试剂盒检测炎症因子的浓度。采用双荧光素酶报告基因法和RNA免疫沉淀法分析miR-370-3p与circ_0035292或TBL1XR1的结合。
结果：Circ_0035292水平在IP患者和LPS触发的WI-38细胞中升高。Circ_0035292敲低拯救了LPS介导的WI-38细胞增殖抑制和WI-38细胞凋亡和炎症促进。Circ_0035292与miR-370-3p相互作用，miR-370-3p直接靶向TBL1XR1。此外，miR-370-3p过表达减轻LPS诱导的WI-38细胞凋亡和炎症损伤,通过TBL1XR1上调而废除。Circ_0035292缺失抑制了NF-κB途径。
结论：circ_0035292的敲低通过miR-370-3p/TBL1XR1轴和NF-κB途径拯救LPS引发的WI-38细胞损伤。
方法：进行定量实时聚合酶链反应和蛋白质印迹检测circ_0035292，microRNA-370-3p（miR-370-3p）和转导蛋白β样1X相关蛋白1（TBL1XR1）的水平。细胞计数试剂盒-8,5-乙炔基-2'-脱氧尿苷,流式细胞术评估细胞增殖和凋亡。用酶联免疫吸附测定试剂盒检测炎症因子的浓度。采用双荧光素酶报告基因法和RNA免疫沉淀法分析miR-370-3p与circ_0035292或TBL1XR1的结合。
结果：Circ_0035292水平在IP患者和LPS触发的WI-38细胞中升高。Circ_0035292敲低拯救了LPS介导的WI-38细胞增殖抑制和WI-38细胞凋亡和炎症促进。Circ_0035292与miR-370-3p相互作用，miR-370-3p直接靶向TBL1XR1。此外，miR-370-3p过表达减轻LPS诱导的WI-38细胞凋亡和炎症损伤,通过TBL1XR1上调而废除。Circ_0035292缺失抑制了NF-κB途径。
结论：circ_0035292的敲低通过miR-370-3p/TBL1XR1轴和NF-κB途径拯救LPS引发的WI-38细胞损伤。
