Effects of Adenosine Triphosphate on Proliferation and Odontoblastic Differentiation of Human Dental Pulp Cells
Effects of Adenosine Triphosphate on Proliferation and Odontoblastic Differentiation of Human Dental Pulp Cells
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DOI：
10.1016/j.joen.2016.07.013
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发表时间：
2016-10-01
影响因子：
4.2
通讯作者：
Xie, Qiufei
中科院分区：
文献类型：
--
作者：
Wang, Wei;Yi, Xiaosong;Xie, Qiufei
腺苷三磷酸（ATP）是一种有效的信号分子，调节细胞中的多种生物活性。它对人牙髓细胞（HDPC）的影响尚不清楚。本研究旨在探讨ATP对HDPC增殖和分化的影响。方法：采用逆转录聚合酶链反应（RT-PCR）技术检测P2受体亚型mRNA的表达。采用流式细胞仪和细胞计数法检测ATP对HDPCs增殖和细胞周期的影响。采用茜素红S染色、能谱分析、Westernblot分析和实时荧光定量聚合酶链反应检测ATP对HDPCs分化的影响。结果：HDPCs中存在P2 X3、P2 X4、P2 X5、P2 X7和所有P2 Y受体亚型。ATP在10 μ mol/L时促进HDPC增殖。浓度.但在800 μ mol/L浓度下，它通过将细胞周期阻滞在G（0）G（1）期而抑制细胞增殖（P <0.05），并诱导成牙本质细胞分化、ERK/MAPK激活以及牙本质基质蛋白1（DMP 1）和牙本质涎磷蛋白（DSPP）mRNA转录。ATP受体拮抗剂苏拉明抑制ERK/MAPK活化和HDPC成牙本质细胞分化（与对照组相比P <0.05）。结论：细胞外ATP激活。诱导HDPC牙源性分化的P2受体和下游信号事件因此，ATP可能通过P2信号促进牙髓组织的愈合和修复。这些结果为牙髓创伤愈合的分子调控提供了新的见解。
Introduction: Adenosine 5'-triphosphate (ATP) is a potent signaling molecule that regulates diverse biological activities in cells. Its effects on human dental pulp cells (HDPCs) remain unknown. This study aimed to examine the effects of ATP on proliferation and differentiation of HDPCs. Methods: Reverse transcription polymerase chain reaction was performed to explore the mRNA expression of P2 receptor subtypes. Cell Counting Kit-8 test and flow cytometry analysis were used to examine the effects of ATP on proliferation and cell cycle of HDPCs. The effects of ATP on differentiation of HDPCs were examined by using alizarin red S staining, energy dispersive x-ray analysis, Western blot analysis, and real-time polymerase chain reaction. Results: The purinoceptors P2X3, P2X4, P2X5, P2X7, and all P2Y receptor subtypes were confirmed to present in HDPCs. ATP enhanced HDPC proliferation at 10 mu mol/L. concentration. However, it inhibited cell proliferation by arresting the cell cycle in G(0)G(1) phase (P < .05 versus control) and induced odontoblastic differentiation, ERK/MAPK activation, and dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) mRNA transcriptions at 800 mu mol/L concentration. Suramin, an ATP receptor antagonist, inhibited ERK/MAPK activation and HDPC odontoblastic differentiation (P < .05 versus control). Conclusions: Extracellular ATP activates. P2 receptors and downstream signaling events that induce HDPC odontogenic differentiation. Thus, ATP may promote dental pulp tissue healing and repair through P2 signaling. Results provide new insights into the molecular regulation of pulpal wound healing.