MicroRNA-182-5p regulates the MAPK/NF-κB pathway by targeting <i>MAPK1</i> to treat acute lung injury

SHEN Xiaoli; ZHUANG Xueming; QIAN Mengshu; MA Weixiong; CHEN Yueyang; LU Chaoming; FANG Xutao; WANG Shibo

doi:10.7619/jcmp.20214224

Journal of Clinical Medicine in Practice > 2022 > 26(5): 79-85. > DOI: 10.7619/jcmp.20214224

SHEN Xiaoli, ZHUANG Xueming, QIAN Mengshu, MA Weixiong, CHEN Yueyang, LU Chaoming, FANG Xutao, WANG Shibo. MicroRNA-182-5p regulates the MAPK/NF-κB pathway by targeting MAPK1 to treat acute lung injury[J]. Journal of Clinical Medicine in Practice, 2022, 26(5): 79-85. DOI: 10.7619/jcmp.20214224

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MicroRNA-182-5p regulates the MAPK/NF-κB pathway by targeting MAPK1 to treat acute lung injury

1.
Department of Emergency, the 904th Hospital of the Joint Logistics Support Force of the Chinese People′s Liberation Army, Wuxi, Jiangsu, 214000
2.
Department of Thoracic Surgery, the 904th Hospital of the Joint Logistics Support Force of the Chinese People′s Liberation Army, Wuxi, Jiangsu, 214000

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Received Date: October 26, 2021
Available Online: March 24, 2022
Published Date: March 14, 2022

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Abstract

Abstract

Objective To explore the effect of miR-182-5p on lipopolysaccharide (LPS) induced acute lung injury (ALI) and its molecular mechanism through molecular biological methods.
Methods Fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-182-5p and MAPK1 in the LPS-induced ALI model. Dual luciferase reporter gene was used to detect the relationship between miR-182-5p and MAPK1. ALI models were treated with Ad-MAPK1, Ad-miR-182-5p and Ad-MAPK1+Ad-miR-182-5p, respectively, and the changes of lung injury index and pulmonary fibrosis were detected by Hematoxylin-eosin staining (HE staining) and Masson staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and myeloperoxidase (MPO) in lung tissues of all treatment groups were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of nuclear factor κB(NF-κB) and NF-κB inhibitor protein α (IκBα) were detected by western blot (WB).
Results This study showed that the expression level of miR-182-5p gradually increased at 0 to 12 hours after ALI, and then decreased at >12 to 24 hours; the expression level of MAPK1 gradually decreased at 0 to 12 hours and increased at >12 to 24 hours. Further genetic interference of ALI showed that the overexpression of miR-182-5p could significantly down-regulate the lung injury index, lung fibrosis and the levels of pro-inflammatory cytokines, while the overexpression of MAPK1 aggravated the levels of lung injury index, lung fibrosis and pro-inflammatory cytokines. Cell experiments in vitro showed that LPS stimulation significantly increased the protein expression of NF-κB and inhibited the expression of IκBα at the same time. The miR-182-5p was able to inhibit the expression of NF-κB and promote the expression of IκBα; on the contrary, the overexpression of MAPK1 reversed this phenomenon.
Conclusion MiR-182-5p plays a therapeutic role in ALI by inhibiting the MAPK/NF-κB signaling pathway, and the data of this study may contribute to an in-depth understanding of the potential molecular mechanism of ALI and the development of new therapeutic methods for ALI.

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References (21)

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MicroRNA-182-5p regulates the MAPK/NF-κB pathway by targeting MAPK1 to treat acute lung injury