MicroRNA‑16 regulates lipopolysaccharide‑induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

Li,Xiaobo; Chu,Qian; Wang,Huaqi

doi:10.3892/etm.2021.10414

MicroRNA‑16 regulates lipopolysaccharide‑induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

Authors:
- Xiaobo Li
- Qian Chu
- Huaqi Wang
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Affiliations: Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China
Published online on: July 12, 2021 https://doi.org/10.3892/etm.2021.10414
Article Number: 982
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Acute lung injury (ALI) is mainly caused by inflammation and is associated with high mortality rates. Emerging evidence has suggested that microRNAs (miRNAs or miRs) serve a significant function in ALI. However, the fundamental mechanism underlying ALI remain to be fully elucidated. Although miR‑16 has been reported to be involved in the occurrence and development of a number of diseases its association with ALI has not been previously investigated. Therefore, the present study aimed to explore the role of miR‑16 in the lipopolysaccharide (LPS)‑induced ALI model. The expression levels of tumor necrosis factor α (TNF‑α), interleukin (IL)‑1β and IL‑6 were measured by ELISA in the blood samples of rats with ALI and in the normal human bronchial epithelial (NHBE) cell line. The role of miR‑16 in inflammation was evaluated using gene overexpression and silencing experiments in NHBE cells by reverse transcription‑quantitative PCR. In addition, the expression levels of inflammatory factors TNF‑α, IL‑1β and IL‑6 were also determined using ELISA. The potential interaction between miR‑16 and TLR4 was assessed using bioinformatics analysis by the TargetScan database and then verified in 293T cells using luciferase reporter assay. The expression of miR‑16 was notably decreased in the lung tissues of rats with LPS‑induced ALI compared with the PBS treated‑group. Additionally, the levels of the proinflammatory cytokines TNF‑α, IL‑1β and IL‑6 were reduced following transfection of NHBE cells with miR‑16 mimics compared with those in the miR‑negative control group. Western blot analysis revealed that miR‑16 overexpression could downregulate TLR4 expression in NHBE cells compared with that in the miR‑NC group. Luciferase reporter assay confirmed that TLR4 may be directly targeted by miR‑16. The effect of miR‑16 on TLR4 was rescued in NHBE cells following treatment with LPS. Overall, these aforementioned findings suggest that miR‑16 may serve a protective role against LPS‑mediated inflammatory responses in NHBE cells by regulating TLR4, where this mechanism may be considered to be a novel approach for treating ALI in the future.

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