Department of Neonatology, Jingzhou Central Hospital (Jingzhou Hospital Affiliated to Yangtze University), Jingzhou, Hubei, China.
Department of Neonatology, Jingzhou Central Hospital (Jingzhou Hospital Affiliated to Yangtze University), Jingzhou, Hubei, China.
Microb Pathog. 2022 Dec;173(Pt A):105819. doi: 10.1016/j.micpath.2022.105819. Epub 2022 Oct 8.
Neonatal pneumonia is a common illness in the neonatal period with a high fatality rate. Accumulating proofs have attested to the crucial role of circular RNAs (circRNAs) in pneumonia. This study was intended to expound on the function of circ_0038467 and the underlying mechanism in lipopolysaccharide (LPS)-stimulated 16HBE cell injury in neonatal pneumonia.
16HBE cells were exposed to LPS to establish an in vitro neonatal pneumonia cell model. Quantitative real-time polymerase chain reaction (qRT-PCR) was implemented for detecting the levels of circ_0038467, microRNA-545-3p (miR-545-3p), and tumor necrosis factor receptor-associated factor 1 (TRAF1) in neonatal pneumonia serums and LPS-treated 16HBE cells. Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) incorporation, and flow cytometry assays were used to examine cell viability, proliferation, and apoptosis, respectively. The protein abundances of proliferation/apoptosis/inflammation-correlated makers and TRAF1 were tested by Western blot. RNase R and Actinomycin D assays were implemented to determine the features of circ_0038467. The mutual effect between miR-545-3p and circ_0038467 or TRAF1 was affirmed by a dual-luciferase reporter and RNA pull-down assay assays.
Circ_0038467 was upregulated in neonatal pneumonia serum specimens and LPS-triggered 16HBE cells. LPS administration restrained 16HBE cell proliferation and promoted apoptosis and inflammation, whereas circ_0038467 silence recovered these influences. Meanwhile, miR-545-3p was targeted by circ_0038467, and circ_0038467 could modulate LPS-treated 16HBE cell injury through absorbing miR-545-3p. Furthermore, circ_0038467 controlled TRAF1 level via segregating miR-545-3p. Moreover, TRAF1 overexpression relieved the suppressive impact of circ_0038467 silence in LPS-triggered 16HBE cell detriment.
Circ_0038467 knockdown mitigated LPS-exposed 16HBE cell damage through regulating miR-545-3p/PPARA axis.
新生儿肺炎是新生儿期的一种常见疾病，死亡率较高。越来越多的证据证明环状 RNA（circRNA）在肺炎中起着关键作用。本研究旨在阐述 circ_0038467 在脂多糖（LPS）刺激的新生儿肺炎 16HBE 细胞损伤中的作用及其潜在机制。
用 LPS 处理 16HBE 细胞，建立体外新生儿肺炎细胞模型。采用实时定量聚合酶链反应（qRT-PCR）检测新生儿肺炎血清和 LPS 处理的 16HBE 细胞中 circ_0038467、微小 RNA-545-3p（miR-545-3p）和肿瘤坏死因子受体相关因子 1（TRAF1）的水平。细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）掺入和流式细胞术分别用于检测细胞活力、增殖和凋亡。Western blot 检测增殖/凋亡/炎症相关标志物和 TRAF1 的蛋白含量。采用 RNase R 和放线菌素 D 测定来确定 circ_0038467 的特征。通过双荧光素酶报告和 RNA 下拉实验证实了 miR-545-3p 与 circ_0038467 或 TRAF1 之间的相互作用。
circ_0038467 在新生儿肺炎血清标本和 LPS 诱导的 16HBE 细胞中上调。LPS 处理抑制 16HBE 细胞增殖，促进细胞凋亡和炎症，而沉默 circ_0038467 则恢复了这些影响。同时，circ_0038467 靶向 miR-545-3p，通过吸收 miR-545-3p 调节 LPS 处理的 16HBE 细胞损伤。此外，circ_0038467 通过分离 miR-545-3p 来控制 TRAF1 水平。此外，TRAF1 过表达减轻了 circ_0038467 沉默在 LPS 诱导的 16HBE 细胞损伤中的抑制作用。
circ_0038467 敲低通过调节 miR-545-3p/PPARA 轴减轻 LPS 暴露的 16HBE 细胞损伤。
