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 Ca2+ ([Ca2+]i), transient cytotoxicity, and induction of receptor activator of nuclear factor kappa-B ligand (RANKL) expression. Intracellular Ca2+ mobilization in human gingival fibroblast-1 (hGF-1) cells was measured using the fluorescent Ca2+ 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 [Ca2+]i 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.

서 론
 실험방법 및 실험재료
  세포 배양 및 시약
  세포독성 실험
  RT-PCR 및 PCR
  통계학적 유의성 검사
 결 과
  Cch 자극 정도에 따른 hGF-1 세포 내 [Ca2+]i증가
  고농도 Cch 자극에 따른 hGF-1 세포 세포독성 확인
  고농도 Cch 자극에 의한 hGF-1 세포 내 골 재형성 관련 유전자 발현
 고 찰
 References

• Seong-Jong Heo(Department of Oral Physiology, and Institute of Biomaterial-Implant, School of Dentistry, Wonkwang University, Iksan 54538, Republic of Korea)
• Min Seuk Kim(Department of Oral Physiology, and Institute of Biomaterial-Implant, School of Dentistry, Wonkwang University, Iksan 54538, Republic of Korea) Corresponding Author