IL-21通过Notch信号调控人牙周膜细胞RANKL表达和增殖的研究

目的:观察IL-21对人牙周膜成纤维细胞RANKL/OPG的表达以及增殖活性的影响,以及Notch信号通路在其中的作用,探讨IL-21在根尖周炎骨破坏中的作用机制。方法:使用不同浓度(0、10、50、100 μg/L)的IL-21作用于人牙周膜成纤维细胞,并选择最佳刺激浓度IL-21(50 μg/L)处理人牙周膜成纤维细胞,观察不同的时间点(0、12、24 h),通过实时定量PCR和ELISA的方法检测细胞中RANKL/OPG的表达水平。同时使用实时定量PCR和Western-blot检测IL-21对人牙周膜成纤维细胞Notch1表达水平的影响。运用Notch信号的阻断剂GSI(5 μmol/L)预处理人牙周膜成纤维细胞后,再检测IL-21对人牙周膜成纤维细胞RANKL/OPG的表达水平的影响。CCK8法测定各个实验组细胞增殖活性。结果:IL-21可显著提高人牙周膜成纤维细胞RANKL的表达水平,对OPG的表达无显著影响。IL-21显著抑制人牙周膜成纤维细胞的增殖活性。结论:IL-21通过Notch信号通路上调人牙周膜成纤维细胞的RANKL表达,抑制其增殖活性。

IL-21 Regulates RANKL Expression and Proliferation Activity in Human Periodontal Ligament Cells via Notch Signal Pathway

Objective: To explore the effect of interleukin-21 (IL-21) via Notch signal pathway on the expression of RANKL/OPG and proliferation in hPDLCs and to investigate the possible role of IL-21 in bone resorption of apical periodontitis. Methods: hPDLCs were treated with IL-21 in different concentrations (0, 10, 25, 50 μg/L). Then the optimal concentration (50μg/L) was used in the time-dependent experiment. The expressions of RANKL/OPG were detected by real-time PCR and ELISA. The expressions of RANKL/OPG were detected by real-time PCR and ELISA. Real-time PCR and western-blot were conducted to detect the expression of Notch1 in hPDLCs treated by IL-21. After pretreatment with GSI (5μmol/L) to inhibit Notch1 signaling pathway, the expressions of RANKL/OPG of hPDLCs treated with IL-21 (50μg/L) were investigated. CCK8 was carried out to detect the proliferation activity in the different groups of hPDLCs. Results: The expressions of RANKL were significantly up-regulated by IL-21. IL-21 decreased the cell proliferation activity in hPDLCs. Conclusion: IL-21 up-regulated the expression of RANKL and reduced proliferation activity in hPDLCs via Notch signal pathway.