He Hai-Peng, Zhao Mei-Zhen, Jiao Wei-Hua, Liu Zhi-Qiang, Zeng Xian-Hai, Li Quan-Li, Hu Tian-Yong, Cheng Bao-Hui
Department of Dentistry, Shenzhen Longgang Otolaryngology hospital & Shenzhen Otolaryngology Research Institute, Shenzhen, 518172, China.
Research Center for Marine Drugs, State Key Laboratory of Microbial Metabolism, Department of Pharmacy, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200127, China.
Stem Cell Res Ther. 2024 Aug 7;15(1):247. doi: 10.1186/s13287-024-03812-2.
The role of periodontal ligament stem cells (PDLSCs) in repairing periodontal destruction is crucial, but their functions can be impaired by excessive oxidative stress (OS). Nocardamine (NOCA), a cyclic siderophore, has been shown to possess anti-cancer and anti-bacterial properties. This study aimed to investigate the protective mechanisms of NOCA against OS-induced cellular dysfunction in PDLSCs.
The cytotoxicity of NOCA on PDLSCs was assessed using a CCK-8 assay. PDLSCs were then treated with hydrogen peroxide (HO) to induce OS. ROS levels, cell viability, and antioxidant factor expression were analyzed using relevant kits after treatment. Small molecule inhibitors U0126 and XAV-939 were employed to block ERK signaling and Wnt pathways respectively. Osteogenic differentiation was assessed using alkaline phosphatase (ALP) activity staining and Alizarin Red S (ARS) staining of mineralized nodules. Expression levels of osteogenic gene markers and ERK pathway were determined via real-time quantitative polymerase chain reaction (RT-qPCR) or western blot (WB) analysis. β-catenin nuclear localization was examined by western blotting and confocal microscopy.
NOCA exhibited no significant cytotoxicity at concentrations below 20 µM and effectively inhibited HO-induced OS in PDLSCs. NOCA also restored ALP activity, mineralized nodule formation, and the expression of osteogenic markers in HO-stimulated PDLSCs. Mechanistically, NOCA increased p-ERK level and promoted β-catenin translocation into the nucleus; however, blocking ERK pathway disrupted the osteogenic protection provided by NOCA and impaired its ability to induce β-catenin nuclear translocation under OS conditions in PDLSCs.
NOCA protected PDLSCs against HO-induced OS and effectively restored impaired osteogenic differentiation in PDLSCs by modulating the ERK/Wnt signaling pathway.
牙周膜干细胞（PDLSCs）在修复牙周组织破坏中起着关键作用，但其功能可能会因过度氧化应激（OS）而受损。诺卡胺（NOCA）是一种环状铁载体，已被证明具有抗癌和抗菌特性。本研究旨在探讨NOCA对OS诱导的PDLSCs细胞功能障碍的保护机制。
使用CCK-8法评估NOCA对PDLSCs的细胞毒性。然后用过氧化氢（HO）处理PDLSCs以诱导OS。处理后使用相关试剂盒分析活性氧（ROS）水平、细胞活力和抗氧化因子表达。分别使用小分子抑制剂U0126和XAV-939阻断ERK信号通路和Wnt通路。通过碱性磷酸酶（ALP）活性染色和矿化结节的茜素红S（ARS）染色评估成骨分化。通过实时定量聚合酶链反应（RT-qPCR）或蛋白质印迹（WB）分析确定成骨基因标志物和ERK通路的表达水平。通过蛋白质印迹和共聚焦显微镜检查β-连环蛋白的核定位。
NOCA在浓度低于20 µM时无明显细胞毒性，并有效抑制HO诱导的PDLSCs中的OS。NOCA还恢复了HO刺激的PDLSCs中的ALP活性、矿化结节形成和成骨标志物的表达。机制上，NOCA增加了p-ERK水平并促进β-连环蛋白向细胞核的转运；然而，阻断ERK通路破坏了NOCA提供的成骨保护，并损害了其在PDLSCs的OS条件下诱导β-连环蛋白核转运的能力。
NOCA通过调节ERK/Wnt信号通路保护PDLSCs免受HO诱导的OS，并有效恢复PDLSCs中受损的成骨分化。
