Lipopolysaccharide‐mediated ATP signaling regulates interleukin‐6 mRNA expression via the P2‐purinoceptor in human dental pulp cells

Author:

Orimoto Ai1ORCID,Kitamura Chiaki1,Ono Kentaro2

Affiliation:

1. Division of Endodontics and Restorative Dentistry Kyushu Dental University Fukuoka Japan

2. Division of Physiology Kyushu Dental University Fukuoka Japan

Abstract

AbstractDental pulp cells play a crucial role in maintaining the balance of the pulp tissue. They actively respond to bacterial inflammation by producing proinflammatory cytokines, particularly interleukin‐6 (IL‐6). While many cell types release adenosine triphosphate (ATP) in response to various stimuli, the mechanisms and significance of ATP release in dental pulp cells under inflammatory conditions are not well understood. This study aimed to investigate ATP release and its relationship with IL‐6 during the inflammatory response in immortalized human dental pulp stem cells (hDPSC‐K4DT) following lipopolysaccharide (LPS) stimulation. We found that hDPSC‐K4DT cells released ATP extracellularly when exposed to LPS concentrations above 10 μg/mL. ATP release was exclusively attenuated by N‐ethylmaleimide, whereas other inhibitors, including clodronic acid (a vesicular nucleotide transporter inhibitor), probenecid (a selective pannexin‐1 channel inhibitor), meclofenamic acid (a selective connexin 43 inhibitor), suramin (a nonspecific P2 receptor inhibitor), and KN‐62 (a specific P2X7 antagonist), did not exhibit any effect. Additionally, LPS increased IL‐6 mRNA expression, which was mitigated by the ATPase apyrase enzyme, N‐ethylmaleimide, and suramin, but not by KN‐62. Moreover, exogenous ATP induced IL‐6 mRNA expression, whereas ATPase apyrase, N‐ethylmaleimide, and suramin, but not KN‐62, diminished ATP‐induced IL‐6 mRNA expression. Overall, our findings suggest that LPS‐induced ATP release stimulates the IL‐6 pathway through P2‐purinoceptor, indicating that ATP may function as an anti‐inflammatory signal, contributing to the maintenance of dental pulp homeostasis.

Funder

Japan Society for the Promotion of Science

Publisher

Wiley

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