中国实用口腔科杂志

中国实用口腔科杂志 ›› 2026, Vol. 19 ›› Issue (4): 458-464.DOI: 10.19538/j.kq.2026.04.012

• 论著 • 上一篇    下一篇

炎症条件下lncRNA-NR_037840.3对牙髓干细胞成骨及成牙向分化的影响初探

师保江a，姬瑞生b，刘苁芮a，曹    钰b   

  1. 首都医科大学附属北京口腔医院a牙体牙髓科，b口腔显微诊疗中心，北京 100070
  • 出版日期:2026-07-30 发布日期:2026-07-30
  • 通讯作者: 曹钰
  • 基金资助:
    北京市自然科学基金（7232070）

  • Online:2026-07-30 Published:2026-07-30

摘要： 目的    初步探讨炎症条件下lncRNA-NR_037840.3对牙髓干细胞（dental pulp stem cells，DPSCs）成骨及成牙向分化的影响，并分析其可能相关的机制。方法    收集2023年12月至2024年12月于首都医科大学附属北京口腔医院就诊患者因正畸治疗需要拔除的前磨牙6颗，取出牙髓组织后行DPSCs分离培养与传代。采用慢病毒转染分别构建稳定敲低和过表达lncRNA-NR_037840.3的DPSCs并进行验证，记为敲低对照组、敲低实验组、过表达对照组、过表达实验组。采用质量浓度10 μg/L肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）刺激作为体外炎症模拟条件。常规成骨及成牙向诱导条件下，第0、3、5、7、14天采用实时荧光定量PCR检测DPSCs中lncRNA-NR_037840.3的表达情况。炎症条件下对4组行成骨及成牙向分化诱导，并行碱性磷酸酶（alkaline phosphatase，ALP）活性检测、茜素红染色、钙沉积量测定，通过Western blot检测牙本质涎磷蛋白（dentin sialophosphoprotein，DSPP）、牙本质基质蛋白1（dentin matrix protein 1，DMP1）、Runt相关转录因子2（Runt-related transcription factor 2，RUNX2）、β-连环蛋白（β-catenin）和磷酸化β-连环蛋白（phosphorylated β-catenin，p-β-catenin）的表达情况。结果    倒置显微镜配套荧光滤光片下观察可见4组明显荧光表达，敲低实验组lncRNA-NR_037840.3相对表达量低于敲低对照组，过表达实验组lncRNA-NR_037840.3相对表达量高于过表达对照组。常规成骨及成牙向诱导条件下，lncRNA-NR_037840.3相对表达量随诱导时间延长总体呈上升趋势。在质量浓度10 μg/L TNF-α刺激下成骨及成牙向分化诱导，与过表达对照组比较，过表达实验组ALP活性升高、相对钙沉积量增加，差异均有统计学意义（t值分别为8.096、23.590，P值分别为0.015、0.002）；其矿化结节形成增多，DSPP、DMP1和RUNX2蛋白表达呈一定增强趋势。与敲低对照组比较，敲低实验组ALP活性降低、相对钙沉积量减少，差异均有统计学意义（t值分别为4.365，6.583，P值分别为0.049、0.022）；其矿化结节形成减少，DSPP、DMP1、RUNX2、β-catenin和p-β-catenin蛋白表达呈一定减弱趋势。结论    在质量浓度10 μg/L TNF-α刺激下，lncRNA-NR_037840.3可促进DPSCs成骨及成牙向分化，其作用可能与β-catenin磷酸化水平变化相关。

关键词: lncRNA-NR_037840.3, 牙髓干细胞, 炎症条件, 成骨及成牙向分化

Abstract: Objective    To preliminarily investigate the effects of lncRNA-NR_037840.3 on the osteogenic and odontogenic differentiation of dental pulp stem cells（DPSCs）under inflammatory conditions，and to explore the potential underlying mechanisms. Methods    Six premolars extracted for orthodontic purposes were collected from patients treated at Beijing Stomatological Hospital，Capital Medical University，between December 2023 and December 2024. Dental pulp tissues were isolated for DPSC culture and passaging. Stable knockdown and overexpression of lncRNA-NR_037840.3 in DPSCs were established by lentiviral transfection and subsequently verified. The cells were assigned to four groups：knockdown control group，knockdown experimental group，overexpression control group，and overexpression experimental group. Stimulation with 10 μg/L tumor necrosis factor-α（TNF-α）was used to establish an in vitro inflammatory condition. Under conventional osteogenic/odontogenic induction conditions，the expression of lncRNA-NR_037840.3 in DPSCs was detected by real-time quantitative PCR on days 0，3，5，7，and 14. Under inflammatory conditions，the four groups were subjected to osteogenic/odontogenic induction，followed by alkaline phosphatase（ALP）activity assay，alizarin red staining，and calcium deposition assay. Western blot was performed to detect the expression of dentin sialophosphoprotein（DSPP），dentin matrix protein 1（DMP1），Runt-related transcription factor 2（RUNX2），β-catenin，and phosphorylated β-catenin（p-β-catenin）. Results    Evident fluorescence signals were observed in all four groups under an inverted microscope equipped with fluorescence filters. The relative expression level of lncRNA-NR_037840.3 was lower in the knockdown experimental group than in the knockdown control group，whereas it was higher in the overexpression experimental group than in the overexpression control group. Under conventional osteogenic/odontogenic induction conditions，the relative expression level of lncRNA-NR_037840.3 showed an overall increasing trend with prolonged induction time. After osteogenic/odontogenic induction under stimulation with 10 μg/L TNF-α，the overexpression experimental group showed significantly higher ALP activity and increased relative calcium deposition compared with the overexpression control group，with statistically significant differences（t = 8.096 and 23.590，respectively；P = 0.015 and 0.002，respectively）. In addition，mineralized nodule formation was increased，and the protein expression levels of DSPP，DMP1，and RUNX2 showed an increasing trend. Compared with the knockdown control group，the knockdown experimental group showed reduced ALP activity and decreased relative calcium deposition，with statistically significant differences（t = 4.365 and 6.583，respectively；P = 0.049 and 0.022，respectively）. In addition，mineralized nodule formation was decreased，and the protein expression levels of DSPP，DMP1，RUNX2，β-catenin，and p-β-catenin showed a decreasing trend. Conclusion    Under stimulation with 10 μg/L TNF-α，lncRNA-NR_037840.3 can promote the osteogenic and odontogenic differentiation of DPSCs，and this effect may be associated with alterations in β-catenin phosphorylation.

Key words: lncRNA-NR_037840.3, dental pulp stem cells, inflammatory condition, osteogenic and odontogenic differentiation