Dong Tao, Sun Xuemin, Jin He
Department of stomatology, BenQ Medical Center, The Affiliated BenQ Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China.
J Periodontol. 2021 Aug;92(8):1192-1200. doi: 10.1002/JPER.20-0176. Epub 2020 Nov 3.
Periodontitis is a chronic inflammatory disease that occurs in periodontal tissues and can cause tooth loosening and loss in severe cases. As the main effector of downstream of Hippo signaling pathway, yes-related protein 1 (YAP1) plays an important role in cell proliferation and differentiation. However, the role of YAP1 in periodontitis has not been reported.
Cell activity was detected by Cell Counting Kit-8 (CCK-8). YAP1 was overexpressed by cell transfection, and then RT-qPCR and western blot were used to detect the expression of YAP1. The cell proliferation was determined by clone formation assay, and the expression of proliferation-related proteins was determined by western blot. The cell differentiation was detected by ELISA kit of alkaline phosphatase activity (ALP) and alizarin red staining. Finally, western blot was used to detect the expression of differentiation-related protein and Hippo signaling pathway-related proteins. Apoptosis was detected by flow cytometry.
With the increase of concentration induced by TNF-α, the cell survival rate of human periodontal ligament stem cells (HPDLSCs) decreased significantly. After the overexpression of YAP1, cell proliferation and proliferation-related protein expression increased. Overexpression of YAP1 can improve the differentiation and the formation of osteoblasts of HPDLSCs induced by TNF-α. The expression of Hippo signaling pathway-related proteins transcriptional coactivators with PDZ binding domains (TAZ), TEA domain family member (TRED) increased and proliferation-related protein P27 decreased, whereas there was no significant change in the expression of MST1.
TNF-α can inhibit proliferation and osteogenic differentiation of HPDLSCs, which can be ameliorated by the YAP1 gene through the Hippo signaling pathway. Our paper suggested that YAP1 may be a potential therapeutic target for periodontitis.
牙周炎是一种发生于牙周组织的慢性炎症性疾病，严重时可导致牙齿松动和脱落。Yes相关蛋白1（YAP1）作为Hippo信号通路下游的主要效应因子，在细胞增殖和分化中发挥重要作用。然而，YAP1在牙周炎中的作用尚未见报道。
采用细胞计数试剂盒-8（CCK-8）检测细胞活性。通过细胞转染过表达YAP1，然后用RT-qPCR和蛋白质免疫印迹法检测YAP1的表达。采用克隆形成试验测定细胞增殖，用蛋白质免疫印迹法测定增殖相关蛋白的表达。通过碱性磷酸酶活性（ALP）ELISA试剂盒和茜素红染色检测细胞分化。最后，用蛋白质免疫印迹法检测分化相关蛋白和Hippo信号通路相关蛋白的表达。采用流式细胞术检测细胞凋亡。
随着TNF-α诱导浓度的增加，人牙周膜干细胞（HPDLSCs）的细胞存活率显著降低。YAP1过表达后，细胞增殖及增殖相关蛋白表达增加。YAP1过表达可改善TNF-α诱导的HPDLSCs的分化和成骨细胞形成。Hippo信号通路相关蛋白PDZ结合结构域转录共激活因子（TAZ）、TEA结构域家族成员（TRED）的表达增加，增殖相关蛋白P27表达降低，而MST1表达无明显变化。
TNF-α可抑制HPDLSCs的增殖和成骨分化，YAP1基因可通过Hippo信号通路改善这种抑制作用。本文提示YAP1可能是牙周炎的一个潜在治疗靶点。
