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Role and mechanism of miR‑144‑5p in LPS‑induced macrophages

  • Authors:
    • Guozhu Zhou
    • Yuwei Li
    • Jianping Ni
    • Ping Jiang
    • Zili Bao
  • View Affiliations / Copyright

    Affiliations: Third Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210000, P.R. China, Department of Orthopedics, Suzhou Hospital of Traditional Chinese Medicine, Suzhou, Jiangsu 215000, P.R. China, Department of Orthopedics, Jingjiang Chinese Medicine Hospital, Taizhou, Jiangsu 214500, P.R. China
    Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 241-247
    |
    Published online on: November 19, 2019
       https://doi.org/10.3892/etm.2019.8218
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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.
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Copy and paste a formatted citation
Spandidos Publications style
Zhou G, Li Y, Ni J, Jiang P and Bao Z: Role and mechanism of miR‑144‑5p in LPS‑induced macrophages. Exp Ther Med 19: 241-247, 2020.
APA
Zhou, G., Li, Y., Ni, J., Jiang, P., & Bao, Z. (2020). Role and mechanism of miR‑144‑5p in LPS‑induced macrophages. Experimental and Therapeutic Medicine, 19, 241-247. https://doi.org/10.3892/etm.2019.8218
MLA
Zhou, G., Li, Y., Ni, J., Jiang, P., Bao, Z."Role and mechanism of miR‑144‑5p in LPS‑induced macrophages". Experimental and Therapeutic Medicine 19.1 (2020): 241-247.
Chicago
Zhou, G., Li, Y., Ni, J., Jiang, P., Bao, Z."Role and mechanism of miR‑144‑5p in LPS‑induced macrophages". Experimental and Therapeutic Medicine 19, no. 1 (2020): 241-247. https://doi.org/10.3892/etm.2019.8218
Copy and paste a formatted citation
x
Spandidos Publications style
Zhou G, Li Y, Ni J, Jiang P and Bao Z: Role and mechanism of miR‑144‑5p in LPS‑induced macrophages. Exp Ther Med 19: 241-247, 2020.
APA
Zhou, G., Li, Y., Ni, J., Jiang, P., & Bao, Z. (2020). Role and mechanism of miR‑144‑5p in LPS‑induced macrophages. Experimental and Therapeutic Medicine, 19, 241-247. https://doi.org/10.3892/etm.2019.8218
MLA
Zhou, G., Li, Y., Ni, J., Jiang, P., Bao, Z."Role and mechanism of miR‑144‑5p in LPS‑induced macrophages". Experimental and Therapeutic Medicine 19.1 (2020): 241-247.
Chicago
Zhou, G., Li, Y., Ni, J., Jiang, P., Bao, Z."Role and mechanism of miR‑144‑5p in LPS‑induced macrophages". Experimental and Therapeutic Medicine 19, no. 1 (2020): 241-247. https://doi.org/10.3892/etm.2019.8218
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