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Updated: Aug 11, 2025
Three-dimensional Inflammatory Human Tissue Equivalents of Gingiva
Published on: April 3, 2018
C5aR antagonist inhibits LPS-induced inflammation in human gingival fibroblasts via NF-κB and MAPK signaling pathways
Yan Chen1,2, Yang Liu3, Hao Li2
1The First Affiliated Hospital of Harbin Medical University, Department of Oral Maxillofacial Surgery, Harbin, Heilongjiang, China.
Objective:
Abnormal complement activation is associated with periodontitis. W54011 is a novel non-peptide C5aR antagonist (C5aRA) that exhibits favorable anti-inflammatory effects in various inflammatory models. However, whether W54011 inhibits periodontitis has not yet been fully elucidated. To address this, we have investigated the probable anti-inflammatory mechanism of W54011 in LPS-treated inflammation in human gingival fibroblasts (HGFs).
Methodology:
HGFs were isolated from healthy gingival tissue samples using the tissue block method and were identified with immunofluorescence staining. The CCK8 assay and reverse transcription-PCR (RT-PCR) were used to select the optimal induction conditions for Lipopolysaccharide (LPS) and C5aRA (according to supplementary data S1, S2 and S3). The levels of inflammatory cytokines, C5aR, and the activation of NF-κB/MAPK signaling pathways were determined by RT-quantitative PCR (RT-qPCR) and Western blotting.
Results:
Immunofluorescence results showed that vimentin and FSP-1 were positive in HGFs and Keratin was negative in HGFs. Immunofluorescence staining demonstrated that C5aRA inhibited LPS-stimulated nuclear translocation of p-p65. RT-qPCR and Western blotting showed that C5aRA reduced the expression of IL-1β, IL-6, TNF-α, C5aR, p-p65, p-IκBα, p-JNK, p-c-JUN, and TLR4 in LPS-induced HGFs.
Conclusion:
These findings suggested that C5aRA attenuated the release of inflammatory cytokines in LPS-induced HGFs by blocking the activation of the NF-κB and MAPK signaling pathways.
Insights
This study shows that C5aR antagonist (C5aRA) reduces inflammation in human gingival fibroblasts by blocking key signaling pathways. This suggests C5aRA may be a potential treatment for periodontitis.
Area of Science:
- Oral biology and immunology
- Inflammation research
- Pharmacology
Background:
- Abnormal complement activation is linked to periodontitis.
- W54011 (C5aRA) is a novel anti-inflammatory agent.
- The effect of W54011 on periodontitis requires further investigation.
Purpose of the Study:
- To investigate the anti-inflammatory mechanism of W54011 in lipopolysaccharide (LPS)-induced human gingival fibroblasts (HGFs).
- To determine if W54011 can inhibit periodontitis-related inflammation.
Main Methods:
- Human gingival fibroblasts (HGFs) were isolated and characterized.
- Optimal LPS and C5aRA conditions were determined using CCK8 and RT-PCR.
- Inflammatory cytokine levels, C5aR expression, and NF-κB/MAPK pathway activation were assessed via RT-qPCR and Western blotting.
Main Results:
- C5aRA inhibited LPS-induced nuclear translocation of p-p65.
- C5aRA reduced the expression of IL-1β, IL-6, TNF-α, C5aR, p-p65, p-IκBα, p-JNK, p-c-JUN, and TLR4.
- Immunofluorescence confirmed HGF identity and C5aRA's effect on p-p65 translocation.
Conclusions:
- C5aRA attenuates inflammatory cytokine release in LPS-induced HGFs.
- The anti-inflammatory effect is mediated by blocking NF-κB and MAPK signaling pathways.
- C5aRA shows potential for managing periodontitis-associated inflammation.
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