| 摘要: |
| 目的 通过c-Jun氨基末端激酶（JNK）特异性抑制剂SP600125干预JNK信号通路，探究JNK通路是否参与低频脉冲电磁场（PEMF）诱导人牙周膜干细胞（hPDLSC）的成骨分化。方法 用低频PEMF（15 Hz、0～3 mT、每间隔12 h辐照1 h）体外辐照hPDLSC，第7天或第14天时通过qPCR检测细胞内Runt相关转录因子2（Runx2）、碱性磷酸酶（ALP）、骨桥蛋白（OPN）、骨钙蛋白（OCN）等成骨相关基因表达水平，评估低频PEMF诱导hPDLSC成骨分化的能力并筛选适宜的磁场作用强度；通过蛋白质印迹法检测低频PEMF刺激下细胞内JNK和磷酸化JNK（p-JNK）蛋白表达水平，qPCR检测不同浓度SP600125干预后细胞内成骨相关基因表达水平的变化，观察JNK通路在PEMF促进hPDLSC成骨分化过程中是否发挥作用。结果 hPDLSC经15 Hz、2.5 mT的低频PEMF辐照后，Runx2、ALP、OPN、OCN等成骨相关基因表达水平高于其他磁场强度分组（P均<0.05）。低频PEMF刺激下明显促进了细胞内JNK、p-JNK蛋白的表达（P均<0.05）；JNK通路抑制后细胞内成骨相关基因表达水平降低，且20、30 μmol/L SP600125对成骨基因表达的抑制效果较10 μmol/L SP600125更明显（P均<0.05）。结论 15 Hz、2.5 mT的PEMF可通过部分激活JNK通路诱导hPDLSC成骨分化。 |
| 关键词: 低频脉冲电磁场 牙周膜干细胞 JNK信号通路 成骨分化 SP600125 |
| DOI：10.16781/j.CN31-2187/R.20220363 |
| 投稿时间：2022-04-30修订日期：2022-11-21 |
| 基金项目:上海市卫生和计划生育委员会科研课题（20134418），解放军总后勤部面上项目（CHJ13J035），海军军医大学（第二军医大学）第一附属医院青年启动基金（2019QN16），海军军医大学（第二军医大学）第一附属医院“234学科攀峰计划”（2020YXK028），海军军医大学（第二军医大学）第一附属医院教改项目（CHJG2020040）. |
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| Role of JNK pathway in osteogenic differentiation of periodontal ligament stem cells stimulated by low-frequency pulsed electromagnetic fields |
| WANG Yan-en1△,WANG Pei2△,WANG Rong1,WEI Yi-bo1,CAO Zhi-zhong1,CHEN Tie-lou1* |
(1. Department of Stomatology, The First Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200433, China;
2. Department of Stomatology, Shanghai 411 Hospital, China Rongtong Medical Healthcare Group Co. Ltd, Shanghai 200081, China
△Co-first authors.
* Corresponding author) |
| Abstract: |
| Objective To explore whether c-Jun N-terminal kinase (JNK) pathway is involved in the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) induced by low-frequency pulsed electromagnetic field (PEMF) via using the specific inhibitor SP600125 to intervene JNK signaling pathway. Methods hPDLSCs were exposed to the low-frequency PEMF stimulation (15 Hz, 0-3.0 mT, radiate for 1 h every 12 h) in vitro for 7 d or 14 d and the expression levels of osteogenic-related genes Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteopontin (OPN) and osteocalcin (OCN) were detected by quantitative polymerase chain reaction (qPCR), so as to determine the osteogenic differentiation ability of cells induced by low-frequency PEMF and the appropriate magnetic field intensity. To determine whether JNK pathway plays a role in the osteogenic differentiation of hPDLSCs stimulated by low-frequency PEMF, the expression levels of JNK and phosphorylated-JNK (p-JNK) proteins were detected by Western blotting; and the expression levels of osteogenic genes in cells treated with different concentrations of SP600125 were detected by qPCR. Results The expression levels of osteogenic-related genes Runx2, ALP, OPN and OCN were higher in hPDLSCs irradiated with low-frequency PEMF at 15 Hz and 2.5 mT than other intensity groups (all P<0.05). Low-frequency PEMF significantly stimulated the expression of JNK and p-JNK proteins in cells (both P<0.05). The expression levels of osteogenic genes in hPDLSCs were decreased after the JNK pathway was inhibited by SP600125, and the inhibitory effects of 20 and 30 μmol/L SP600125 were more obvious than that of 10 μmol/L SP600125 (both P<0.05). Conclusion PEMF (15 Hz,2.5 mT) partially activates JNK pathway to induce hPDLSC osteogenic differentiation. |
| Key words: low-frequency pulsed electromagnetic fields periodontal ligament stem cells JNK signaling pathway osteogenic differentiation SP600125 |
