低强度脉冲超声对人牙周膜细胞Hippo信号通路的影响  

Effects of Low Intensity Pulsed Ultrasound on Hippo Signaling Pathway in Human Periodontal Membrane Cells

作　　者：李平[1] 尹娟[1] 徐旻馨 薛慧 LI Ping;YIN Juan;XU Minxin;XUE Hui(Departments of Stomatology and Central Laboratory,The Affiliated Suzhou Hospital of Nanjing Medical University,Suzhou Municipal Hospital,Gusu School,Nanjing Medical University,Suzhou 215008,China)

机构地区：[1]南京医科大学姑苏学院,南京医科大学附属苏州医院,苏州市立医院中心实验室,苏州市立医院口腔科,江苏苏州215008

出　　处：《口腔医学研究》2022年第11期1087-1091,共5页Journal of Oral Science Research

基　　金：苏州民生科技项目(编号:SYSD2020245、SYS2020177);第五批姑苏卫生分层培养青年拔尖人才项目(编号:GSWS2019062、GSWS2020077);苏州市科教兴卫青年科技项目(编号:KJXW2021039)。

摘　　要：目的:观察低强度脉冲超声(low intensity pulsed ultrasound,LIPUS)对人牙周膜细胞(human periodontal ligament cells,hPDLCs)Hippo信号通路相关基因表达的影响。方法:原代培养hPDLCs,对第3代hPDLCs进行30 mW/cm^(2)、45 mW/cm^(2)、60 mW/cm^(2)3个辐照强度的LIPUS刺激25 min,连续刺激3 d。Western blot及定量聚合酶链反应(Q-PCR)法检测Hippo信号通路成骨相关基因的表达,观察细胞形态变化,筛选最佳的刺激条件。对刺激21 d的细胞进行茜素红染色,与对照组相比分析LIPUS对成骨的影响。结果:本研究通过鉴定成功培养出原代hPDLCs,LIPUS刺激hPDLCs后Hippo信号通路YAP、TAZ及RUNX2基因与蛋白表达增强,磷酸化的TAZ(p-TAZ)及YAP(p-YAP)蛋白表达下调,并且45 mW/cm^(2)与60 mW/cm^(2)成骨效果较30 mW/cm^(2)更加明显。hPDLCs经LIPUS(60 mW/cm^(2))刺激21 d后,钙结节明显较对照组增多。结论:适宜强度的低脉冲超声能够促进hPDLCs Hippo信号通路成骨相关基因的表达,这为优化牙齿正畸环境提供一定的理论基础。Objective:To observe the effects of low intensity pulsed ultrasound(LIPUS)on the genes expression of Hippo signaling pathway in human periodontal ligament cells(hPDLCs).Methods:The primary cells were cultured,and the third generation of periodontal cells were stimulated with LIPUS for 3 days with the intensity of 30 mW/cm^(2),45 mW/cm^(2),and 60 mW/cm^(2) for 25 min.Western blot and Q-PCR were used to detect the genes expression of Hippo signaling pathway.The changes of cell morphology were observed,and the optimal treatment conditions were screened.Alizarin red staining was performed on cells stimulated for 21 days to analyze the effect of LIPUS on osteogenesis compared with the control group.Results:In this study,primary periodontal membrane cells were successfully cultured.The expression of YAP,TAZ,and RUNX2 in Hippo signaling pathway was significantly enhanced,and the p-TAZ and p-YAP expressions were down-regulated in periodontal membrane cells with LIPUS stimulated.The osteogenic effect of 45 mW/cm^(2) and 60 mW/cm^(2) was more obvious than that of 30 mW/cm^(2).In contrast to the control group,calcium nodules increased significantly after LIPUS(60 mW/cm^(2))stimulation.Conclusion:Low-pulse ultrasound with appropriate intensity can promote the expression of osteogenic genes of Hippo signaling pathway in periodontal membrane cells,which can provide a theoretical basis to optimize the orthodontic environment.

关 键 词：牙齿正畸 低脉冲超声 人牙周膜细胞 Hippo信号通路 

分 类 号：R783.5[医药卫生—口腔医学]