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The negative feedback regulation of microRNA-146a in human periodontal ligament cells after Porphyromonas gingivalis lipopolysaccharide stimulation

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Abstract

Objective

Toll-like receptors (TLRs) pathway has been demonstrated to play an important role in periodontitis. However, the regulatory mechanism of microRNAs (miRNAs) on TLRs pathway is still unclear. Hence, this study is to explore the function of miRNA-146a in inflammatory reaction induced by Porphyromonas gingivalis lipopolysaccharide (LPS) in human periodontal ligament cells (hPDLCs).

Methods

Cells were treated with 1 or 10 μg/ml P. gingivalis LPS. The expression of TLR2, TLR4 and miRNA-146a were measured by real-time polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was applied to detect nuclear factor (NF)-κ B p65 nuclear activity, interleukin-1β (IL-1β), IL-6, IL-8 and tumor necrosis factor-α (TNF-α). To examine the underlying mechanisms, cells were exposed to anti-TLR2/4 mAb or miRNA-146a inhibitor/mimic and evaluated by real-time PCR and ELISA.

Results

10 μg/ml P. gingivalis LPS increased the expressions of TLR2 (3.79 ± 0.31), TLR4 (2.21 ± 0.31), and miRNA-146a (4.91 ± 0.87), NF-κ B p65 nuclear activity (6.51 ± 0.77 fold) (p < 0.05). 1 μg/ml P. gingivalis LPS induced TLR2 (3.05 ± 0.23), miRNA-146a (3.66 ± 0.83) and NF-κ B p65 nuclear activity (4.06 ± 0.78 fold) (p < 0.05), except TLR4 (1.11 ± 0.30, p > 0.05). Also, cytokines production increased (p < 0.05). The up-regulation of miRNA-146a could be blocked by anti-TLR2/4 mAb (p < 0.05). After the blockage of miRNA-146a, TLR2, TLR4, NF-κ B p65 nuclear activity and proinflammatory cytokines increased. However, after application of miRNA-146a mimic, the levels of these indexes decreased obviously (p < 0.05).

Conclusion

MiRNA-146a functions as a negative feedback regulator via down-regulating proinflammatory cytokine secretion and blocking TLRs signaling pathway in hPDLCs after P. gingivalis LPS stimulation.

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Acknowledgments

This work was supported by Science and Technology Development Project of Tianjin Education Commission (2010134) and National Natural Science Foundation (81100758/H1405).

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Authors and Affiliations

  1. Department of Periodontics, Institute of Stomatology, School of Dentistry, Hospital of Stomatology, Tianjin Medical University, Tianjin, 300070, China

    Shao-Yun Jiang, Dong Xue, Yun-Yun Dong, Cong-Cong Wei & Jia-Yin Deng

  2. Department of Periodontology, Shanghai Research Institute of Stomatology, Ninth People’s Hospital, Shanghai Key Laboratory of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China

    Yu-Feng Xie

  3. Department of Oral Surgery, Institute of Stomatology, School of Dentistry, Hospital of Stomatology, Tianjin Medical University, Tianjin, 300070, China

    Dong-Wang Zhu

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  1. Shao-Yun Jiang
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Correspondence to Shao-Yun Jiang.

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Jiang, SY., Xue, D., Xie, YF. et al. The negative feedback regulation of microRNA-146a in human periodontal ligament cells after Porphyromonas gingivalis lipopolysaccharide stimulation. Inflamm. Res. 64, 441–451 (2015). https://doi.org/10.1007/s00011-015-0824-y

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  • DOI: https://doi.org/10.1007/s00011-015-0824-y

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