miRNA-146 negatively regulates the production of pro-inflammatory cytokines via NF-κB signalling in human gingival fibroblasts

Journal of Inflammation

Zhi-kai Lin¹  Jia-chen Dong¹  Qiu-man Guo¹  Zhong-chen Song¹  Shao-yun Jiang¹  Rong Shu¹  Yu-feng Xie¹ 

[1] Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology, Shanghai, China

关键词: NF-κB;    Pro-inflammatory cytokines;    Human gingival fibroblasts;    miRNA-146;   

Others  :  1140712
DOI  :  10.1186/s12950-014-0038-z

 received in 2014-01-23, accepted in 2014-11-15,  发布年份 2014

【 摘 要 】

Objective

In human gingival fibroblasts (HGFs), TLR4 recognises Pathogen-associated molecular patterns (PAMPs), such as LPS, and subsequently activates downstream signals that lead to the production of pro-inflammatory cytokines. The aim of this study was to explore the mechanisms of LPS-induced miRNA-146 regulation of TLR4 signals in HGFs.

Materials and methods

HGFs were treated with Porphyromonas gingivalis (P.g) LPS, the cells were harvested, and kinase phosphorylation levels were detected by western blot. Selective pharmacological inhibitors and agonists were used to block or activate the relevant kinases, miRNA-146a/b expression levels were detected by real-time PCR, and IL-1, IL-6, and TNF-α production were measured by enzyme-linked immunosorbent assays (ELISA). A luciferase reporter plasmid containing miRNA-146a/b promoter was tested in terms of p50/p65 regulation.

Results

After P.g LPS treatment, NF-κB and Erk1/2 were strongly activated in HGFs. miRNA-146a/b, IL-1, IL-6 and TNF-α levels were down-regulated when NF-κB inhibitor was used. p50/p65 strongly activated miRNA-146a/b promoter as measured with the luciferase assay.

Conclusion

In TLR4 signalling in HGFs, both miRNA-146a and miRNA-146b are downstream targets of NF-κB, but not of AP-1 signalling. miRNA-146a/b expression was specifically dependent on NF-κB but not Erk1/2 or JNK signalling.

【 授权许可】

   
2014 Xie et al.; licensee BioMed Central.

【 图 表 】

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