JNK通路在低频脉冲电磁场促进牙周膜干细胞成骨分化中的作用  

Role of JNK pathway in osteogenic differentiation of periodontal ligament stem cells stimulated by low-frequency pulsed electromagnetic fields

作　　者：王彦蒽 汪沛 王蓉 蔚一博[1] 曹志中[1] 陈铁楼 WANG Yan-en;WANG Pei;WANG Rong;WEI Yi-bo;CAO Zhi-zhong;CHEN Tie-lou(Department of Stomatology,The First Affiliated Hospital of Naval Medical University(Second Military Medical University),Shanghai 200433,China;Department of Stomatology,Shanghai 411 Hospital,China Rongtong Medical Healthcare Group Co.Ltd,Shanghai 200081,China)

机构地区：[1]海军军医大学(第二军医大学)第一附属医院口腔科,上海200433 [2]中国融通医疗健康集团有限公司上海四一一医院口腔科,上海200081

出　　处：《海军军医大学学报》2023年第3期283-291,共9页Academic Journal of Naval Medical University

基　　金：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)。

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、30μmol/L SP600125对成骨基因表达的抑制效果较10μmol/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。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 30μmol/L SP600125 were more obvious than that of 10μmol/L SP600125(both P<0.05).Conclusion PEMF(15 Hz,2.5 mT)partially activates JNK pathway to induce hPDLSC osteogenic differentiation.

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 

分 类 号：R783.1[医药卫生—口腔医学]