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Circ_0001498通过与miR-574-5p相互作用上调SOX6,促进脂多糖诱导的脓毒症相关急性肺损伤中的肺细胞凋亡和炎症。

Circ_0001498 contributes to lipopolysaccharide-induced lung cell apoptosis and inflammation in sepsis-related acute lung injury via upregulating SOX6 by interacting with miR-574-5p.

作者信息

Hu Wei, Wang Qin, Luo Zhichun, Shi Yaqiong, Zhang Liangping, Zhang Zhijun, Liu Jianlin, Liu Kelan

机构信息

Intensive care unit, Liyang People's Hospital, Liyang, China.

出版信息

Gen Physiol Biophys. 2023 Jan;42(1):37-47. doi: 10.4149/gpb_2022054.

Abstract

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.

摘要

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。

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