Song Zhi, Chen Lingling, Guo Jia, Qin Wei, Wang Runfu, Huang Shuheng, Yang Xiaoting, Tian Yaguang, Lin Zhengmei
Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, China; Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong, China.
Department of Stomatology, Hainan General Hospital, Haikou, Hainan, China.
J Endod. 2017 Feb;43(2):315-320. doi: 10.1016/j.joen.2016.10.021. Epub 2016 Dec 29.
Calcium ions (Ca) actively participate in reparative dentin formation by promoting cellular proliferation and differentiation of human dental pulp cells (hDPCs). Transient receptor potential cation channel, subfamily C, member 1 (TRPC1) activates Ca entry upon store depletion in a variety of cell types. However, the function of TRPC1 in hDPCs has not been reported. Therefore, we aimed to analyze the role of TRPC1 in hDPCs undergoing odontoblast-like differentiation.
Immunohistochemical staining was used to determine the distribution of TRPC1 in pulp tissues. Western blot analysis was used to detect the protein level of TRPC1 in the odontoblast-like differentiation of hDPCs. Knockdown of TRPC1 was performed with an adenoviral vector to evaluate the role of TRPC1 in hDPCs during odontoblast-like differentiation.
The results showed that TRPC1 was highly expressed in the cytoplasm of dental pulp cells, especially in the odontoblast layer of the healthy pulp. Moreover, the protein level of TRPC1 increased in a time-dependent manner during the odontoblast-like differentiation of hDPCs. Importantly, knockdown of TRPC1 attenuated the process of odontoblast-like differentiation as indicated by the reduction in mineralized nodules and the down-regulation of dentin sialophosphoprotein and dentin matrix protein 1. Moreover, knockdown of TRPC1 decreased Ca entry to the cytoplasm of hDPCs.
Our data indicated a pivotal role of TRPC1 in the odontoblastlike differentiation of hDPCs, which may be a therapeutic target to enhance reparative dentin formation.
钙离子(Ca)通过促进人牙髓细胞(hDPCs)的细胞增殖和分化,积极参与修复性牙本质的形成。瞬时受体电位阳离子通道C亚家族成员1(TRPC1)在多种细胞类型中,于储存耗尽时激活钙离子内流。然而,TRPC1在hDPCs中的功能尚未见报道。因此,我们旨在分析TRPC1在hDPCs向成牙本质细胞样分化过程中的作用。
采用免疫组织化学染色法确定TRPC1在牙髓组织中的分布。利用蛋白质印迹分析检测hDPCs向成牙本质细胞样分化过程中TRPC1的蛋白水平。使用腺病毒载体敲低TRPC1,以评估TRPC1在hDPCs向成牙本质细胞样分化过程中的作用。
结果显示,TRPC1在牙髓细胞的细胞质中高表达,尤其是在健康牙髓的成牙本质细胞层。此外,在hDPCs向成牙本质细胞样分化过程中,TRPC1的蛋白水平呈时间依赖性增加。重要的是,敲低TRPC1可减弱成牙本质细胞样分化过程,表现为矿化结节减少以及牙本质涎磷蛋白和牙本质基质蛋白1表达下调。此外,敲低TRPC1可减少钙离子进入hDPCs的细胞质。
我们的数据表明TRPC1在hDPCs向成牙本质细胞样分化中起关键作用,这可能是增强修复性牙本质形成的一个治疗靶点。