Peng Yu-Zhi, Tang Song-Jiang, Li Xiao-Jing, Zhao Sheng-Ke, Liu Bao-Zhen
First Affiliated Hospital of Guiyang University of Traditional Chinese Medicine.Guiyang 550001, Guizhou Province, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2020 Aug;29(4):370-374.
To investigate the role of P2X7 receptor (P2X7r) in osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs).
hPDLSCs were isolated from the premolars collected in the First Affiliated Hospital of Guiyang University of Traditional Chinese Medicine, and divided into four groups. Group A was cultured in conventional medium, group B was cultured in osteogenic induction medium, group C was cultured in osteogenic induction medium + 100 nmol/L adenosine triphosphate (ATP) solution, and group D was cultured in osteogenic induction medium + 100 nmol/L P2X7 receptor specific antagonist KN-62. After 7 days, alizarin red staining was used to observe the osteogenic effect of hPDLSCs in each group. The mRNA expression of osteocalcin (OCN), RUNX2 and P2X7r in hPDLSCs was detected by real-time PCR reaction (RT-PCR). The data were processed by SPSS 22.0 software package.
Alisarin red staining showed that the morphology of hPDLSCs cells in group B and group C was significantly changed. The pale calcified nodules in group C were significantly more than those in group B, while very few calcified nodules were found in group A and group D. The mRNA expression of OCN, RUNX2 and P2X7r in hPDLSCs were the highest in group C, followed by group B(P<0.05), and no difference was found between group A and group D(P>0.05).
P2X7 receptor can promote osteogenic differentiation of human periodontal ligament stem cells after being activated by ATP, which may provide a new direction for clinical treatment of periodontitis.
探讨P2X7受体（P2X7r）在人牙周膜干细胞（hPDLSCs）成骨分化中的作用。
从贵阳中医学院第一附属医院收集的前磨牙中分离出hPDLSCs，并分为四组。A组在常规培养基中培养，B组在成骨诱导培养基中培养，C组在成骨诱导培养基+100 nmol/L三磷酸腺苷（ATP）溶液中培养，D组在成骨诱导培养基+100 nmol/L P2X7受体特异性拮抗剂KN-62中培养。7天后，采用茜素红染色观察各组hPDLSCs的成骨效果。通过实时PCR反应（RT-PCR）检测hPDLSCs中骨钙素（OCN）、RUNX2和P2X7r的mRNA表达。数据采用SPSS 22.0软件包进行处理。
茜素红染色显示，B组和C组hPDLSCs细胞形态发生明显改变。C组淡染的钙化结节明显多于B组，而A组和D组钙化结节极少。hPDLSCs中OCN、RUNX2和P2X7r的mRNA表达在C组最高，其次为B组（P<0.05），A组和D组之间无差异（P>0.05）。
P2X7受体被ATP激活后可促进人牙周膜干细胞的成骨分化，这可能为牙周炎的临床治疗提供新的方向。
