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P 2 X 7 receptor– P annexin1 interaction mediates stress‐induced interleukin‐1 beta expression in human periodontal ligament cells
Author(s) -
Kanjanamekanant K.,
Luckprom P.,
Pavasant P.
Publication year - 2014
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/jre.12139
Subject(s) - pannexin , microbiology and biotechnology , receptor , chemistry , periodontal fiber , small interfering rna , biology , gap junction , connexin , biochemistry , medicine , transfection , intracellular , dentistry , gene
Background and Objective Pannexin 1 (Panx1) has been found to form nonjunctional hemichannels. It is also proposed to combine with the P2X7 receptor, forming a complex involved in adenosine triphosphate ( ATP) ‐induced interleukin‐1beta ( IL ‐1β) release in macrophages. Previously, we reported that mechanical stress induced IL ‐1β expression via the ATP /P2X7 receptor‐dependent pathway in human periodontal ligament ( HPDL ) cells and that ATP was released through the connexin 43 (Cx43) hemichannel. In the present work, we examined the role of P anx1 in stress‐induced IL ‐1β induction in HPDL cells. Material and Methods Cultured HPDL cells were treated with compressive loading or ATP to stimulate IL ‐1β expression. Inhibitors, antagonists and the small interfering RNA technique were used to investigate the involvement of P anx1 in IL ‐1β induction. Co‐immunoprecipitation (Co‐ IP ) and immunostaining were used to determine the association of Panx1 with the P 2X7 receptor. The IL ‐1β release mechanism was analyzed using inhibitors. Results Blocking Panx1 significantly decreased ATP release, as well as IL ‐1β up‐regulation, upon stimulation with stress or ATP . Co‐ IP revealed the association of Panx1 and the P2X7 receptor in HPDL cells, which was increased in response to mechanical loading. Pretreatment with vesicular trafficking inhibitors significantly reduced the amount of IL ‐1β released from stimulated cells, suggesting that IL ‐1β might be released through vesicles. Conclusion We clearly illustrated the contribution of P anx1 in ATP release, as well as in IL ‐1β induction in HPDL cells. The association of P anx1 and the P2X7 receptor might be required for IL ‐1β induction, and their possible novel role in IL ‐1β vesicular release was indicated.