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Published on: May 31, 2018
NEAT1 Promotes LPS-induced Inflammatory Injury in Macrophages by Regulating MiR-17-5p/TLR4
Yanhui Li1, Wei Guo2, Yeping Cai2
1ChenZhou NO.1 People's Hospital LuoJiaJin, ChenZhou China.
Background:
The inflammatory response of macrophages is responsible for sepsis. Long noncoding RNA nuclear enriched abundant transcript 1 (NEAT1) has been reported to be involved in sepsis development. However, its underlying mechanism remains largely unclear. This study aims to investigate the effect of NEAT1 on inflammatory response of macrophages and explore the regulatory network of NEAT1/microRNA-17-5p (miR-17-5p)/Toll-like receptor 4 (TLR4).
Methods:
The serum samples of 68 sepsis patients and 32 heathy controls were collected. THP-1 macrophages were treated with lipopolysaccharide (LPS) to induce inflammatory injury model of sepsis. The expressions of NEAT1, miR-17-5p and TLR4 were measured by quantitative real-time polymerase chain reaction or western blot. The inflammatory response was investigated by levels of inflammatory cytokines, tumor necrosis factor-alpha (TNF-ɑ), interleukin-1beta (IL-1β) and IL-6 as well as nitric oxide (NO) production. The interaction among NEAT1, miR-17-5p and TLR4 were investigated by bioinformatics analysis, luciferase reporter assay and RNA pull-down.
Results:
NEAT1 expression was enhanced in patient serum and associated with severity of sepsis. Knockdown of NEAT1 inhibited levels of TNF-ɑ, IL-1β, IL-6 and NO release in LPS-treated macrophages. miR-17-5p is bound to NEAT1 and its abrogation reversed NEAT1 knockdown-mediated inhibition of inflammatory response in LPS-treated macrophages. Overexpression of miR-17-5p weakened LPS-induced inflammatory response. TLR4 as a target of miR-17-5p was regulated by NEAT1 and miR-17-5p. TLR4 res-to ration alleviated silencing NEAT1-induced inflammatory suppression.
Conclusion:
Silence of NEAT1 suppressed LPS-induced inflammatory response of macrophages by mediating miR-17-5p and TLR4, indicating that NEAT1 might be a promising target for sepsis treatment.
Insights
Long noncoding RNA NEAT1 promotes sepsis by increasing inflammatory responses in macrophages. Silencing NEAT1 reduces inflammation via the microRNA-17-5p/TLR4 pathway, suggesting NEAT1 as a potential sepsis therapeutic target.
Area of Science:
- Molecular Biology
- Immunology
- Genetics
Background:
- Sepsis pathogenesis involves macrophage inflammatory responses.
- Long noncoding RNA NEAT1 is implicated in sepsis development, but its mechanism is unclear.
- This study investigates NEAT1's role in macrophage inflammation and the NEAT1/microRNA-17-5p/Toll-like receptor 4 regulatory network.
Purpose of the Study:
- To elucidate the mechanism of NEAT1 in sepsis-induced macrophage inflammation.
- To explore the regulatory network involving NEAT1, microRNA-17-5p (miR-17-5p), and Toll-like receptor 4 (TLR4).
Main Methods:
- Collected serum from sepsis patients and healthy controls.
- Induced a sepsis inflammatory injury model in THP-1 macrophages using lipopolysaccharide (LPS).
- Quantified NEAT1, miR-17-5p, and TLR4 expression; measured inflammatory cytokines (TNF-ɑ, IL-1β, IL-6) and nitric oxide (NO); utilized bioinformatics, luciferase reporter assays, and RNA pull-down assays.
Main Results:
- Elevated NEAT1 expression in sepsis patients correlated with disease severity.
- NEAT1 knockdown suppressed LPS-induced inflammatory cytokine and NO production in macrophages.
- NEAT1 regulates miR-17-5p, which targets TLR4; this interaction mediates the inflammatory response.
- Restoring TLR4 expression reversed the anti-inflammatory effects of NEAT1 silencing.
Conclusions:
- NEAT1 silencing mitigates LPS-induced macrophage inflammation by modulating the miR-17-5p/TLR4 axis.
- NEAT1 represents a potential therapeutic target for sepsis treatment.
