Kim Se-Min, Lee Bin-Na, Koh Jeong-Tae, Chang Hoon-Sang, Hwang In-Nam, Oh Won-Mann, Min Kyung-San, Hwang Yun-Chan
Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University, Youngbong-ro 77, Buk-gu, Gwangju, 61186, Korea.
Department of Pharmacology and Dental Therapeutics, School of Dentistry, Dental Science Research Institute, Chonnam National University, Youngbong-ro 77, Buk-gu, Gwangju, 61186, Korea.
BMC Oral Health. 2017 May 26;17(1):89. doi: 10.1186/s12903-017-0379-0.
Chlormadinone acetate (CMA) is a derivative of progesterone and is used as an oral contraceptive. The aim of this study was to investigate the effects of CMA on odontogenic differentiation and mineralization of human dental pulp cells (hDPCs) and related signaling pathways.
Cell viability was determined by the water-soluble tetrazolium (WST)-1 assay. Odontogenic differentiation of hDPCs was evaluated by real-time polymerase chain reaction using odontogenic marker genes, such as alkaline phosphatase (ALP), osteocalcin (OCN), dentin sialophosphoprotein (DSPP), and dentin matrix protein-1 (DMP-1). Mineralization of hDPCs was evaluated by ALP staining and alizarin red staining. The extracellular signal-regulated kinase (ERK) pathway was examined by Western blot analysis.
There was no statistically significant difference in cell viability between the control and CMA-treated groups. Our analysis of odontogenic marker genes indicated that CMA enhanced the expression of those genes. CMA-treated hDPCs showed increased ALP activity and formation of mineralized nodules, compared with control-treated cells. In addition, CMA stimulation resulted in phosphorylation of ERK and resulted in inhibition of downstream molecules by the ERK inhibitor U0126.
These findings suggest that CMA improves odontogenic differentiation and mineralization of hDPCs through the ERK signaling pathway.
醋酸氯地孕酮(CMA)是一种孕激素衍生物,用作口服避孕药。本研究的目的是探讨CMA对人牙髓细胞(hDPCs)牙源性分化和矿化以及相关信号通路的影响。
采用水溶性四氮唑(WST)-1法测定细胞活力。通过使用牙源性标记基因,如碱性磷酸酶(ALP)、骨钙素(OCN)、牙本质涎磷蛋白(DSPP)和牙本质基质蛋白-1(DMP-1)的实时聚合酶链反应来评估hDPCs的牙源性分化。通过ALP染色和茜素红染色评估hDPCs的矿化情况。通过蛋白质印迹分析检测细胞外信号调节激酶(ERK)通路。
对照组和CMA处理组之间的细胞活力无统计学显著差异。我们对牙源性标记基因的分析表明,CMA增强了这些基因的表达。与对照组处理的细胞相比,CMA处理的hDPCs显示出ALP活性增加和矿化结节形成。此外,CMA刺激导致ERK磷酸化,并导致ERK抑制剂U0126对下游分子的抑制。
这些发现表明,CMA通过ERK信号通路改善hDPCs的牙源性分化和矿化。