Int J Oral Biol.  2016 Sep;41(3):119-123. 10.11620/IJOB.2016.41.3.119.

Activation of acetylcholine receptor elicits intracellular Ca2+ mobilization, transient cytotoxicity, and induction of RANKL expression

• Heo SJ
• Kim MS

• KMID: 2354199
• DOI: http://doi.org/10.11620/IJOB.2016.41.3.119

Abstract

Acetylcholine receptors (AChR) including muscarinic and nicotinic AChR are widely expressed and mediate a variety of physiological cellular responses in neuronal and non-neuronal cells. Notably, a functional cholinergic system exists in oral epithelial cells, and nicotinic AChR (nAChR) mediates cholinergic anti-inflammatory responses. However, the pathophysiological roles of AChR in periodontitis are unclear. Here, we show that activation of AChR elicits increased cytosolic Ca²âº ([Ca²âº]áµ¢), transient cytotoxicity, and induction of receptor activator of nuclear factor kappa-B ligand (RANKL) expression. Intracellular Ca²âº mobilization in human gingival fibroblast-1 (hGF-1) cells was measured using the fluorescent Ca²âº indicator, fura-2/AM. Cytotoxicity and induction of gene expression were evaluated by measuring the release of glucose-6-phosphate dehydrogenase and RT-PCR. Activation of AChR in hGF-1 cells by carbachol (Cch) induced [Ca²âº]áµ¢ increase in a dose-dependent manner. Treatment with a high concentration of Cch on hGF-1 cells caused transient cytotoxicity. Notably, treatment of hGF-1 cells with Cch resulted in upregulated RANKL expression. The findings may indicate potential roles of AChR in gingival fibroblast cells in bone remodeling.

Keyword

acetylcholine receptor; gingival fibroblast; intracellular Ca²⁺ mobilization; RANKL; Osteoprotegerin

MeSH Terms

Acetylcholine*
Bone Remodeling
Carbachol
Cytosol
Epithelial Cells
Fibroblasts
Gene Expression
Glucosephosphate Dehydrogenase
Humans
Neurons
Osteoprotegerin
Periodontitis
Receptors, Cholinergic
Acetylcholine
Carbachol
Glucosephosphate Dehydrogenase
Osteoprotegerin
Receptors, Cholinergic