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Cl-amidine attenuates lipopolysaccharide-induced inflammation in human gingival fibroblasts via the JNK/MAPK, NF-κB, and Nrf2 signalling pathways

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A Correction to this article was published on 15 February 2023

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Abstract

Cl-amidine has been reported to have anti-inflammatory properties in a variety of diseases. However, the role of Cl-amidine in periodontal disease remains unclear. Here, the purpose of this study was to investigate the effect of Cl-amidine on lipopolysaccharide (LPS)-induced inflammation in human gingival fibroblasts (HGFs). The cytotoxic effect of Cl-amidine was measured with the Cell Counting Kit-8 (CCK-8) assay and Annexin V-FITC/PI staining. The protein levels of IL-6 and IL-8 in culture supernatants were measured with enzyme-linked immunosorbent assay (ELISA). The mRNA levels of inflammatory cytokines, TLR4 and MyD88 were assessed by quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The expression patterns of IL-6, TNF-ɑ, and IL-1β in HGFs were tested with western blot. The levels of NF-κB, MAPK, and Nrf2 pathway-related proteins were detected by western blot. Immunofluorescence (IF) staining was used to examine the nuclear translocation of NF-κB p65. Moreover, a rat gingivitis model was established to further clarify the role of Cl-amidine. Our results showed that Cl-amidine suppressed LPS-induced gingival inflammation both in vitro and in vivo. Mechanistically, Cl-amidine inhibited LPS-induced MyD88 expression, NF-κB activation, and JNK phosphorylation. Additionally, Cl-amidine upregulated Nrf2 and Ho-1 expression both with and without LPS stimulation but did not alter ROS levels or Keap1 expression. Overall, our data suggest that Cl-amidine acts as an inhibitor of LPS-induced gingival inflammation via the JNK/MAPK, NF-κB, and Nrf2 signalling pathways.

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Acknowledgements

This work was supported by the grants from the Natural Science Foundation of Shandong Province (No. ZR2021MC126).

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Author notes

  1. Linlin Zheng and Xiangqin Qi have contributed equally to this work.

Authors and Affiliations

  1. Department of Stomatology, Zibo Central Hospital, Zibo, China

    Jianxin Du & Ning Wang

  2. Department of Cardiac Intensive Care Unit, Zibo Central Hospital, Zibo, China

    Hui Sun

  3. Department of Plastic Surgery, Zibo Central Hospital, Zibo, China

    Linlin Zheng

  4. Department of Ultrasonography, Zibo Central Hospital, No 54 Gongqingtuan West Road, Zibo, 255036, China

    Xiangqin Qi

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Correspondence to Xiangqin Qi.

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Ethical consent was granted from the Ethics Committee of Zibo Central Hospital for studies involving patients. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Ethical approval of the animal study was granted by the Animal Ethics Committee of Zibo Central Hospital.

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Du, J., Wang, N., Sun, H. et al. Cl-amidine attenuates lipopolysaccharide-induced inflammation in human gingival fibroblasts via the JNK/MAPK, NF-κB, and Nrf2 signalling pathways. Human Cell 36, 223–233 (2023). https://doi.org/10.1007/s13577-022-00822-1

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