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Downregulation of lncRNA NEAT1 Ameliorates LPS-Induced Inflammatory Responses by Promoting Macrophage M2 Polarization via miR-125a-5p/TRAF6/TAK1 Axis

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Abstract

The lncRNA nuclear enriched abundant transcript 1 (NEAT1) promotes sepsis-inflammatory responses and acute kidney injury (AKI), but little known about the underlying mechanisms. This study aims to investigate the roles of NEAT1 in regulating macrophage polarization and its potential for alleviating inflammatory responses during sepsis pathogenesis. Mouse RAW264.7 macrophages were treated with lipopolysaccharide (LPS) as a cellular inflammatory model. NEAT1 shRNA, miR-125a-5p mimics, and TRAF6-overexpressing vector were used to transfect RAW264.7 cells. NEAT1, miR-125a-5p, and mRNA levels of functional genes were detected by quantitative RT-PCR. Protein abundances were analyzed by western blotting. Macrophage polarization was evaluated by flow cytometry. The bindings of miR-125a-5p with NEAT1 or TRAF6 gene were validated by dual luciferase reporter assay. LPS treatment promoted NEAT1 and suppressed miR-125a-5p expression in mouse macrophage cells. NEAT1 silencing by shRNAs promoted macrophage M2 polarization under LPS treatment, which upregulated miR-125a-5p expression, repressed TRAF6 expression and TAK1 protein phosphorylation in macrophages. These cellular and molecular changes induced by NEAT1 shRNAs were abrogated by miR-125a-5p inhibitors. Moreover, miR-125a-5p mimics suppressed TRAF6 expression and TAK1 protein phosphorylation in LPS-treated macrophages, thus causing macrophage M2 polarization under LPS treatment. TRAF6 overexpression abrogated the miR-125a-5p mimics-induced macrophage M2 polarization. miR-125a-5p could directly bind to NEAT1 or TRAF6 gene in macrophages. lncRNA NEAT1 knockdown ameliorates LPS-induced inflammation by promoting macrophage M2 polarization via miR-125a-5p/TRAF6/TAK1 axis.

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Abbreviations

AAM:

Alternatively activated macrophage

AKI:

Acute kidney injury

AMPK:

AMP-activated protein kinase

Arg-1:

Arginase-1

ATAD2:

ATPase family AAA domain-containing protein 2

ceRNA:

Competing endogenous RNA

E2F3:

E2F transcription factor 3

iNOS:

Inducible nitric oxide synthase

IL-6:

Interleukin-6;

KLF13:

Krueppel-like factor 13

lncRNAs:

Long non-coding RNAs

LPS:

Lipopolysaccharide

miRNAs:

MicroRNAs

NEAT1:

Nuclear enriched abundant transcript 1

NF-κB:

Nuclear factor-kappa B

PBMCs:

Peripheral blood mononuclear cells

TAK1:

Transforming growth factor-activated kinase 1

TRAF6:

Tumor necrosis factor receptor–associated factor 6

TLR:

Toll-like receptor

TNF-α:

Tumor necrosis factor-alpha

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Authors and Affiliations

  1. Department of Medical Intensive Care Unit, General Hospital of Southern Theater Command, The First School of Clinical Medicine, Southern Medical University, PLA, No.111, Liuhua Road, Guangzhou, 510515, Guangdong Province, People’s Republic of China

    Wei Wang & Zhen-Hui Guo

  2. Department of Emergency, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, 570311, People’s Republic of China

    Wei Wang

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Wang, W., Guo, ZH. Downregulation of lncRNA NEAT1 Ameliorates LPS-Induced Inflammatory Responses by Promoting Macrophage M2 Polarization via miR-125a-5p/TRAF6/TAK1 Axis. Inflammation 43, 1548–1560 (2020). https://doi.org/10.1007/s10753-020-01231-y

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