Hu Wei, Wang Qin, Luo Zhichun, Shi Yaqiong, Zhang Liangping, Zhang Zhijun, Liu Jianlin, Liu Kelan
Intensive care unit, Liyang People's Hospital, Liyang, China.
Gen Physiol Biophys. 2023 Jan;42(1):37-47. doi: 10.4149/gpb_2022054.
Circular RNAs (circRNAs) have important regulation in in sepsis-related acute lung injury (ALI). Circ_0001498 was significantly overexpressed in sepsis-induced acute respiratory distress syndrome. The aims of this study were to explore role and mechanism of circ_0001498 in lipopolysaccharide (LPS)-treated WI-38 cells. Human samples were collected from 56 sepsis patients and 46 healthy volunteers at Liyang People's Hospital. Circ_0001498, microRNA-574-5p (miR-574-5p) or sex-determining region Y-related high-mobility-group box 6 (SOX6) levels were detected via reverse transcription-quantitative polymerase chain reaction assay. Cell viability was determined through Cell Counting Kit-8 assay. Apoptosis rate was examined by flow cytometry. Western blot was used for measurement of proteins. Inflammatory cytokines were detected via enzyme-linked immunosorbent assay. Target relation was analyzed via dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Circ_0001498 was overexpressed in sepsisrelated ALI patients and LPS-treated WI-38 cells. Silencing circ_0001498 reduced LPS-induced cell apoptosis and inflammation. Circ_0001498 interacted with miR-574-5p. The regulation of circ_0001498 knockdown was abolished by miR-574-5p inhibitor. Furthermore, miR-574-5p directly targeted SOX6 and circ_0001498 upregulated SOX6 via targeting miR-574-5p. Overexpression of miR-574-5p alleviated LPS-induced cell injury by downregulating SOX6. This research identified that circ_0001498 facilitated sepsis-related ALI progression by targeting miR-574-5p to upregulate SOX6.
环状RNA(circRNAs)在脓毒症相关急性肺损伤(ALI)中具有重要调控作用。Circ_0001498在脓毒症诱导的急性呼吸窘迫综合征中显著过表达。本研究旨在探讨circ_0001498在脂多糖(LPS)处理的WI-38细胞中的作用及机制。在溧阳市人民医院收集了56例脓毒症患者和46例健康志愿者的人体样本。通过逆转录-定量聚合酶链反应检测Circ_0001498、微小RNA-574-5p(miR-574-5p)或性别决定区Y相关高迁移率族蛋白盒6(SOX6)水平。通过细胞计数试剂盒-8检测法测定细胞活力。通过流式细胞术检测凋亡率。采用蛋白质印迹法检测蛋白质。通过酶联免疫吸附测定法检测炎性细胞因子。通过双荧光素酶报告基因检测法和RNA免疫沉淀(RIP)检测法分析靶标关系。Circ_0001498在脓毒症相关ALI患者和LPS处理的WI-38细胞中过表达。沉默Circ_0001498可减少LPS诱导的细胞凋亡和炎症。Circ_0001498与miR-574-5p相互作用。miR-574-5p抑制剂消除了Circ_0001498敲低的调控作用。此外,miR-574-5p直接靶向SOX6,且Circ_0001498通过靶向miR-574-5p上调SOX6。miR-574-5p的过表达通过下调SOX6减轻LPS诱导的细胞损伤。本研究确定Circ_0001498通过靶向miR-574-5p上调SOX6促进脓毒症相关ALI进展。