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Updated: Jan 1, 2026
Bone Marrow-derived Macrophage Production
Published on: November 22, 2013
Role and mechanism of miR-144-5p in LPS-induced macrophages
Guozhu Zhou1,2, Yuwei Li3, Jianping Ni2
1Third Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210000, P.R. China.
Abstract:
The aim of the present study was to explore the possible role of microRNA-144-5p (miR-144-5p) in rheumatoid arthritis (RA) and the associated mechanism. Following the induction of THP-1 cell differentiation into macrophages by phorbol ester (100 ng/ml) treatment, an in vitro inflammatory model of RA was established by treating the THP-1 macrophages with 1 µg/ml lipopolysaccharide (LPS). The level of miR-144-5p was subsequently measured using reverse transcription-quantitative PCR, which found that the expression of miR-144-5p was significantly reduced in LPS-treated THP-1 macrophages. Bioinformatics analysis and a dual-luciferase reporter assay were used to predict and confirm TLR2 as a direct target of miR-144-5p, respectively. Toll-like receptor 2 (TLR2) was then validated as a target gene of miR-144-5p. The effects of miR-144-5p upregulation and TLR2 silencing on LPS-treated THP-1 macrophages were then determined by transfection with miR-144-5p mimic and TLR2-siRNA, respectively. Cell viability was subsequently measured using a Cell Counting Kit-8 assay, whilst the expression of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-8 secreted by THP-1 macrophages was measured using ELISA. Western blotting was performed to measure p65 phosphorylation (p-p65) in the NK-κB signaling pathway. It was found that miR-144-5p overexpression reduced macrophage cell viability, reduced the expression of TNF-α, IL-6 and IL-8, and reduced the expression of TLR2 and p-p65 compared with the control group. Likewise, TLR2 silencing also reduced macrophage cell viability and reduced the expression of TNF-α, IL-6 and IL-8 in THP-1 macrophages. In conclusion, the data from the present study suggested that miR-144-5p overexpression reduced THP-1 macrophage cell viability and inhibited the expression of TNF-α, IL-6 and IL-8 in cells, possibly by inhibiting the expression of TLR2 and suppressing the activation of NK-κB signaling. Therefore, miR-144-5p may serve as a novel therapeutic target for the treatment of RA.
Insights
MicroRNA-144-5p (miR-144-5p) levels decrease in rheumatoid arthritis (RA) macrophages. Upregulating miR-144-5p or silencing Toll-like receptor 2 (TLR2) reduces inflammation and cell viability, suggesting miR-144-5p as a potential RA therapeutic target.
Area of Science:
- Molecular Biology
- Immunology
- Rheumatology
Background:
- Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by joint inflammation.
- Macrophages play a critical role in RA pathogenesis.
- MicroRNAs (miRNAs) are implicated in regulating inflammatory responses in RA.
Purpose of the Study:
- To investigate the role of microRNA-144-5p (miR-144-5p) in rheumatoid arthritis (RA).
- To elucidate the underlying mechanism of miR-144-5p in RA pathogenesis.
- To explore miR-144-5p as a potential therapeutic target for RA.
Main Methods:
- Established an in vitro RA model using lipopolysaccharide (LPS)-treated THP-1 macrophages.
- Quantified miR-144-5p expression using reverse transcription-quantitative PCR.
- Identified Toll-like receptor 2 (TLR2) as a direct target of miR-144-5p via bioinformatics and dual-luciferase reporter assays.
- Assessed the effects of miR-144-5p mimic and TLR2-siRNA transfection on cell viability, inflammatory cytokine expression (TNF-α, IL-6, IL-8), and NF-κB signaling pathway activation (p-p65).
Main Results:
- LPS treatment significantly reduced miR-144-5p expression in THP-1 macrophages.
- miR-144-5p overexpression decreased macrophage cell viability and suppressed the release of TNF-α, IL-6, and IL-8.
- miR-144-5p upregulation also reduced TLR2 expression and NF-κB pathway activation (p-p65).
- Silencing TLR2 mimicked the effects of miR-144-5p overexpression, reducing cell viability and inflammatory cytokine production.
Conclusions:
- miR-144-5p plays an inhibitory role in RA by targeting TLR2 and suppressing NF-κB signaling.
- Overexpression of miR-144-5p reduces macrophage inflammatory responses and cell viability.
- miR-144-5p represents a promising therapeutic target for rheumatoid arthritis treatment.
