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A role for the calcium-sensing receptor in the expression of inflammatory mediators in LPS-treated human dental pulp cells

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Abstract

The aim of this study is to investigate the role of calcium-sensing receptor (CaSR) in the expression of inflammatory mediators of lipopolysaccharide (LPS)-treated human dental pulp cells (hDPCs). The expression profile of CaSR in LPS-simulated hDPCs was detected using immunofluorescence, real time quantitative PCR (RT-qPCR), and Western blot analyses. Then, its regulatory effects on the expression of specific inflammatory mediators such as interleukin (IL)-1β, IL-6, cyclooxygenase 2 (COX2)-derived prostaglandin E2 (PGE2), tumor necrosis factor (TNF)-α, and IL-10 were determined by RT-qPCR and enzyme-linked immunosorbent assay (ELISA). LPS significantly downregulated the gene expression of CaSR, but upregulated its protein expression level in hDPCs. Treatments by CaSR agonist R568 or its antagonist Calhex231, and their combinations with protein kinase B (AKT) inhibitor LY294002 showed obvious effects on the expression of selected inflammatory mediators in a time-dependent manner. Meanwhile, an opposite direction was found between the action of R568 and Calhex231, as well as the expression of the pro- (IL-1β, IL-6, COX2-derived PGE2, and TNF-α) and anti-inflammatory (IL-10) mediators. The results provide the first evidence that CaSR-phosphatidylinositol-3 kinase (PI3K)-AKT-signaling pathway is involved in the release of inflammatory mediators in LPS-treated hDPCs, suggesting that the activation or blockade of CaSR may provide a novel therapeutic strategy for the treatment of pulp inflammatory diseases.

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Data availability

The datasets used and/or analyzed during the current study available from the corresponding author on reasonable request.

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Funding

This study was supported by the Guangdong Basic and Applied Basic Research Foundation (Grant No. 2019A1515010072), and National Science Foundation of China (Grant No. 81700957).

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Authors and Affiliations

  1. Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, No. 56 Lingyuan Xi Road, Guangzhou, 510055, Guangdong, People’s Republic of China

    Shaofeng An, Yanhuo Chen, Ting Yang, Yihua Huang & Yiwei Liu

  2. Guangdong Province Key Laboratory of Stomatology, Sun Yat-Sen University, No. 74 Zhongshan Er Road, Guangzhou, 510080, Guangdong, People’s Republic of China

    Shaofeng An, Yanhuo Chen, Ting Yang, Yihua Huang & Yiwei Liu

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  1. Shaofeng An
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  2. Yanhuo Chen
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  3. Ting Yang
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  4. Yihua Huang
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  5. Yiwei Liu
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Contributions

YC, YL and TY performed the experiments and analyzed the data. SA and YH designed and supervised this project jointly. SA analyzed the data, drafted the first manuscript and revised the manuscript finally.

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Correspondence to Shaofeng An.

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The authors deny any conflict of interest related to this study.

Ethical approval

All experimental protocols were approved by the Ethics Committee of Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, China (No. KQEC-2022-18-01).

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An, S., Chen, Y., Yang, T. et al. A role for the calcium-sensing receptor in the expression of inflammatory mediators in LPS-treated human dental pulp cells. Mol Cell Biochem 477, 2871–2881 (2022). https://doi.org/10.1007/s11010-022-04486-1

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