环状SLCO3A1缺失通过miR-424-5p/HMGB3途径减轻脂多糖诱导的人肺泡上皮细胞炎症和凋亡。
CircSLCO3A1 depletion ameliorates lipopolysaccharide-induced inflammation and apoptosis of human pulmonary alveolar epithelial cells through the miR-424-5p/HMGB3 pathway.
作者信息
Li Yan, Zhang Chunmei, Zhao Zhongyan
机构信息
Department of Emergency Medicine, China-Japan Union Hospital of Jilin University, Changchun, Jilin China.
Department of Critical Medicine, China-Japan Union Hospital of Jilin University, No. 126, Xiantai Street, Changchun, 130033 Jilin China.
出版信息
Mol Cell Toxicol. 2023 May 25:1-12. doi: 10.1007/s13273-023-00341-6.
BACKGROUND
Recent studies have shown the pathogenesis of acute lung injury (ALI) involves circular RNA (circRNA). However, there are no data on the role of circSLCO3A1 in ALI and the underlying mechanism.
OBJECTIVE
ALI-like cell injury was induced by stimulating human pulmonary alveolar epithelial cells (HPAEpiCs) using lipopolysaccharide (LPS). The expression of circSLCO3A1, miR-424-5p and high mobility group box 3 (HMGB3) was detected by quantitative real-time polymerase chain reaction. Cell viability and cell apoptosis were assessed by cell counting kit-8 (CCK-8) assay and flow cytometry analysis, respectively. Enzyme-linked immunosorbent assay was performed to determine the production of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) and monocyte chemotactic protein 1 (MCP-1). Caspase-3 activity was detected by caspase-3 activity assay. Protein expression of inducible NOS (iNOS), cyclooxygenase-2 (COX2), p-p65 and p65 was analyzed by Western blot. The interactions among circSLCO3A1, miR-424-5p and HMGB3 were identified by dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA pull-down assay.
RESULTS
CircSLCO3A1 and HMGB3 expression were significantly increased, while miR-424-5p was decreased in LPS-treated HPAEpiCs and the serum of septic ALI patients in comparison with controls. CircSLCO3A1 knockdown assuaged LPS-induced HPAEpiC inflammation and apoptosis. Besides, circSLCO3A1 targeted miR-424-5p and regulated LPS-triggered HPAEpiC inflammation and apoptosis by binding to miR-424-5p. Under the treatment of LPS, miR-424-5p mediated HPAEpiC disorders by targeting HMGB3. Importantly, circSLCO3A1 modulated HMGB3 production by interacting with miR-424-5p.
CONCLUSION
CircSLCO3A1 absence assuaged LPS-induced HPAEpiC inflammation and apoptosis through the miR-424-5p/HMGB3 axis.
HIGHLIGHTS
CircSLCO3A1 expression was upregulated in LPS-induced HPAEpiCs and sepsis-induced ALI patients.CircSLCO3A1 depletion protected against LPS-induced HPAEpiC disorders.CircSLCO3A1 bound to miR-424-5p in HPAEpiCs.MiR-424-5p targeted HMGB3 in HPAEpiCs.CircSLCO3A1 regulated HMGB3 expression through miR-424-5p.
SUPPLEMENTARY INFORMATION
The online version contains supplementary material available at 10.1007/s13273-023-00341-6.
背景
近期研究表明，急性肺损伤（ALI）的发病机制涉及环状RNA（circRNA）。然而，关于circSLCO3A1在ALI中的作用及潜在机制尚无相关数据。
目的
使用脂多糖（LPS）刺激人肺泡上皮细胞（HPAEpiCs）诱导ALI样细胞损伤。通过定量实时聚合酶链反应检测circSLCO3A1、miR-424-5p和高迁移率族蛋白B3（HMGB3）的表达。分别采用细胞计数试剂盒-8（CCK-8）法和流式细胞术分析评估细胞活力和细胞凋亡。进行酶联免疫吸附测定以确定白细胞介素-1β（IL-1β）、IL-6、肿瘤坏死因子-α（TNF-α）和单核细胞趋化蛋白1（MCP-1）的产生。通过caspase-3活性测定检测caspase-3活性。采用蛋白质印迹法分析诱导型一氧化氮合酶（iNOS）、环氧化酶-2（COX2）、p-p65和p65的蛋白表达。通过双荧光素酶报告基因测定、RNA免疫沉淀测定和RNA下拉测定确定circSLCO3A1、miR-424-5p和HMGB3之间的相互作用。
结果
与对照组相比，LPS处理的HPAEpiCs和脓毒症ALI患者血清中circSLCO3A1和HMGB3表达显著增加，而miR-424-5p表达降低。敲低circSLCO3A1可减轻LPS诱导的HPAEpiC炎症和凋亡。此外，circSLCO3A1靶向miR-424-5p并通过与miR-424-5p结合调节LPS触发的HPAEpiC炎症和凋亡。在LPS处理下，miR-424-5p通过靶向HMGB3介导HPAEpiC功能紊乱。重要的是，circSLCO3A1通过与miR-424-5p相互作用调节HMGB3的产生。
结论
circSLCO3A1缺失通过miR-424-5p/HMGB3轴减轻LPS诱导的HPAEpiC炎症和凋亡。
要点
circSLCO3A1在LPS诱导的HPAEpiCs和脓毒症诱导的ALI患者中表达上调。circSLCO3A1缺失可预防LPS诱导的HPAEpiC功能紊乱。circSLCO3A1在HPAEpiCs中与miR-424-5p结合。miR-424-5p在HPAEpiCs中靶向HMGB3。circSLCO3A1通过miR-424-5p调节HMGB3表达。
补充信息
在线版本包含可在10.1007/s13273-023-00341-6获取的补充材料。
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