Fang Xing, Mei Wenjing, Zeng Rihua, Zou Li, Zeng Xuefei, Tang Shanghong
Department of PICU, Huizhou Central People's Hospital, Huizhou, Guangdong, China.
Shock. 2023 May 1;59(5):820-828. doi: 10.1097/SHK.0000000000002111. Epub 2023 Mar 5.
Background: Infantile pneumonia is a respiratory infection disease, seriously threatening the life of neonatal patients. Circular RNA (circRNA) dysregulation is reported to be involved in pneumonia pathogenesis. Circ_0012535 was previously displayed to be upregulated in blood samples of patients with community-acquired pneumonia. However, circ_0012535's role in this disorder remains unclear. We thus aim to unveil the functions of circ_0012535 in infantile pneumonia. Methods: Fetal lung fibroblasts (WI38) treated with LPS were used as pneumonia cell models. Expression analysis for circ_0012535, miR-338-3p and IL6R was performed using quantitative real-time polymerase chain reaction. Cell counting kit 88), 5-ethynyl-2'-deoxyuridine, and flow cytometry assays were implemented for cell function detection. The release of inflammatory factors, and superoxide dismutase activity and malonaldehyde content were ascertained using commercial kits. The putative binding between miR-338-3p and circ_0012535 or IL6R was validated by dual-luciferase analysis, RIP analysis, and pull-down analysis. Results: Circ_0012535 was highly expressed in LPS-treated WI38 cells. Knockdown of circ_0012535 recovered LPS-inhibited cell viability and proliferation and attenuated LPS-induced cell apoptosis, cell cycle arrest, inflammation, and oxidative stress. Circ_0012535 bound to miR-338-3p and negatively regulated miR-338-3p expression. Inhibition of miR-338-3p reversed the role of circ_0012535 knockdown, thereby recovering LPS-induced WI38 cell apoptosis and inflammation. MiR-338-3p bound to IL6R 3'UTR, and circ_0012535 shared miR-338-3p binding site with IL6R. IL6R overexpression reversed the role of miR-338-3p, thereby recovering LPS-induced WI38 cell apoptosis and inflammation. Conclusion: Circ_0012535 supported LPS-induced WI38 cell apoptosis and inflammation to promote the progression of infantile pneumonia, and circ_0012535 functioned partly by targeting the miR-338-3p/IL6R signaling.
婴儿肺炎是一种呼吸道感染疾病，严重威胁新生儿患者的生命。据报道，环状RNA（circRNA）失调参与肺炎发病机制。先前显示circ_0012535在社区获得性肺炎患者的血液样本中上调。然而，circ_0012535在这种疾病中的作用仍不清楚。因此，我们旨在揭示circ_0012535在婴儿肺炎中的功能。方法：用脂多糖（LPS）处理的胎儿肺成纤维细胞（WI38）作为肺炎细胞模型。使用定量实时聚合酶链反应对circ_0012535、miR-338-3p和IL6R进行表达分析。采用细胞计数试剂盒88）、5-乙炔基-2'-脱氧尿苷和流式细胞术检测细胞功能。使用商业试剂盒测定炎性因子的释放、超氧化物歧化酶活性和丙二醛含量。通过双荧光素酶分析、RNA免疫沉淀分析和下拉分析验证miR-338-3p与circ_0012535或IL6R之间的假定结合。结果：circ_0012535在LPS处理的WI38细胞中高表达。敲低circ_0012535可恢复LPS抑制的细胞活力和增殖，并减轻LPS诱导的细胞凋亡、细胞周期阻滞、炎症和氧化应激。circ_0012535与miR-338-3p结合并负调节miR-338-3p表达。抑制miR-338-3p可逆转circ_0012535敲低的作用，从而恢复LPS诱导的WI38细胞凋亡和炎症。miR-338-3p与IL6R 3'非翻译区结合，circ_0012535与IL6R共享miR-338-3p结合位点。IL6R过表达可逆转miR-338-3p的作用，从而恢复LPS诱导的WI38细胞凋亡和炎症。结论：circ_0012535支持LPS诱导的WI38细胞凋亡和炎症，促进婴儿肺炎的进展，并且circ_0012535部分通过靶向miR-338-3p/IL6R信号发挥作用。
