Abstract：
Circular RNAs (circRNAs) have been confirmed to mediate infantile pneumonia development. In this, we investigated the role and new mechanism of circ_0035292 regulating infantile pneumonia progression. Lipopolysaccharide (LPS)-treated WI-38 cells were used to mimic infantile pneumonia cell injury models. Quantitative real-time PCR was used to measure circ_0035292, microRNA (miR)-494-3p and toll-like receptor 4 (TLR4). Cell proliferation and apoptosis were assessed by MTT assay, EdU assay, and flow cytometry. Protein expression was tested using western blot analysis. Inflammation and oxidative stress were evaluated by measuring IL-6, IL-1β, MDA and SOD levels using ELISA assay and corresponding kits. RNA interaction was confirmed by dual-luciferase reporter assay and RIP assay. Circ_0035292 had elevated expression in infantile pneumonia patients and LPS-induced WI-38 cells. Silenced circ_0035292 could enhance WI-38 cell proliferation, while suppress apoptosis, inflammation and oxidative stress under LPS treatment. Mechanically, circ_0035292 targeted miR-494-3p to positively regulate TLR4. The rescue experiments indicated that miR-494-3p inhibitor abolished the function of circ_0035292 knockdown, and TLR4 overexpression reversed the inhibitory effect of miR-494-3p on LPS-induced WI-38 cell injury. Circ_0035292 might be a potential target for infantile pneumonia treatment, which knockdown could relieve LPS-induced cell injury via the regulation of miR-494-3p/TLR4 axis.
摘要：
环状RNA(circularRNAs,circRNAs)已被证实介导婴儿肺炎的发展。在此,我们研究了circ_0035292调节小儿肺炎进展的作用和新机制。脂多糖(LPS)处理的WI-38细胞用于模拟婴儿肺炎细胞损伤模型。定量实时PCR用于测量circ_0035292、微小RNA(miR)-494-3p和toll样受体4(TLR4)。MTT法检测细胞增殖和凋亡,EdU分析，和流式细胞术。使用蛋白质印迹分析测试蛋白质表达。炎症和氧化应激通过测量IL-6，IL-1β，使用ELISA测定和相应试剂盒的MDA和SOD水平。通过双荧光素酶报告基因测定和RIP测定确认RNA相互作用。Circ_0035292在婴儿肺炎患者和LPS诱导的WI-38细胞中具有升高的表达。沉默的circ_0035292可以增强WI-38细胞的增殖，在抑制细胞凋亡的同时，LPS处理下的炎症和氧化应激。机械上,circ_0035292靶向miR-494-3p以正向调节TLR4。拯救实验表明，miR-494-3p抑制剂消除了circ_0035292敲低的功能，TLR4过表达逆转了miR-494-3p对LPS诱导的WI-38细胞损伤的抑制作用。Circ_0035292可能是婴儿肺炎治疗的潜在目标，该基因敲低可以通过调节miR-494-3p/TLR4轴减轻LPS诱导的细胞损伤。
