ABSTRACT

This study aimed to identify the role of lncRNA TUG1 with miR-221-3p on mice with lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS). Animal model was established, and lung tissue histopathologic status and permeability were detected by hematoxylin-eosin (HE) or Evans blue dye assay respectively. Levels of inflammation cytokines, lncRNA TUG1, miR-221-3p, sprouty related EVH1 domain-containing 2 (SPRED2), and phosphorylated (p)-ERK1/2 were determined by ELISA, qRT-PCR or Western blot. Pulmonary impairment and apoptosis were examined by flow cytometry. We observed that LPS up-regulated levels of tumor necrosis factor-α (TNF-α), Interleukin-1β (1L-1β), and ERK1/2 phosphorylation, and reduced SPRED2 levels, which were rescued by overexpressed lncRNA TUG1. StarBase and dual-luciferase reporter assay verified that miR-221-3p was targeted by lncRNA TUG1. MiR-221-3p could reverse the effect of lncRNA TUG1 on cell apoptosis, levels of TNF-α, IL-1β, SPRED2, and p-ERK1/2. Therefore, overexpressed lncRNA TUG1 attenuated LPS-induced pulmonary impairment in ARDS mice via regulating miR-221-3p/SPRED2 axis.

摘要

这项研究旨在确定具有miR-221-3p的lncRNA TUG1在脂多糖（LPS）诱导的急性呼吸窘迫综合征（ARDS）小鼠中的作用。建立动物模型，分别用苏木精-伊红（HE）或伊文思蓝染料法检测肺组织的病理状态和通透性。通过ELISA，qRT-PCR或Western blot测定炎症细胞因子，lncRNA TUG1，miR-221-3p，含发芽相关EVH1域的2（SPRED2）和磷酸化（p）-ERK1 / 2的水平。通过流式细胞术检查肺损伤和细胞凋亡。我们观察到LPS上调了肿瘤坏死因子-α（TNF-α），白细胞介素-1β（1L-1β）和ERK1 / 2磷酸化水平，并降低了SPRED2水平，这可以通过过量表达的lncRNA TUG1来挽救。StarBase和双荧光素酶报告基因检测证实了miR-221-3p被lncRNA TUG1靶向。MiR-221-3p可以逆转lncRNA TUG1对细胞凋亡，TNF-α，IL-1β，SPRED2和p-ERK1 / 2的影响。因此，过表达的lncRNA TUG1通过调节miR-221-3p / SPRED2轴减轻了ARDS小鼠中LPS诱导的肺损伤。