上海口腔医学

上海口腔医学 ›› 2019, Vol. 28 ›› Issue (1): 6-12.doi: 10.19439/j.sjos.2019.01.002

• 论著 • 上一篇    下一篇

力生长因子对周期性牵张力诱导的人牙周膜细胞成骨分化及MMP-1、MMP-2表达的影响

陈静涛1, 王燕1, 周志斐2, 卫克文1   

  1. 1.空军军医大学第二附属医院 口腔科,陕西 西安 710038;
    2.中国人民解放军西藏军区总医院 口腔科,西藏 拉萨 850000
  • 收稿日期:2018-04-02 修回日期:2018-06-26 出版日期:2019-02-25 发布日期:2019-04-12
  • 通讯作者: 卫克文,E-mail: weikewentd@163.com
  • 作者简介:陈静涛（1984-）,男,硕士,主治医师,E-mail: cjt1984_td@163.com
  • 基金资助:
    陕西省自然科学基金（2017Jc2-06）

Mechano-growth factor regulated cyclic stretch-induced osteogenic differentiation and MMP-1, MMP-2 expression in human periodontal ligament cells by activating the MEK/ERK1/2 pathway

CHEN Jing-tao1, WANG Yan1, ZHOU Zhi-fei2, WEI Ke-wen1   

  1. 1.Department of Stomatology, the Second Affiliated Hospital of Air Force Military Medical University. Xi'an 710038, Shaanxi Province;
    2.Department of Stomatology, General Hospital of Tibet Military Region of the Chinese People's Liberation Army. Lhasa 850000, Tibet, China
  • Received:2018-04-02 Revised:2018-06-26 Online:2019-02-25 Published:2019-04-12

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摘要： 目的:探讨力生长因子（mechano-growth factor,MGF）对周期性牵张力(cyclic stretch,CS）作用下人牙周膜细胞成骨分化及MMP1、MMP-2表达的影响及其作用机制。方法:分离培养人牙周膜细胞 (human periodontal ligament cells, hPDLCs),将si-MGF转染细胞,用MGF刺激或MEK/EKR通路抑制剂U0126进行干预,以Flexercell应力加载系统对细胞施加形变率10%、频率0.1 Hz的周期性牵张应力;采用碱性磷酸酶(alkaline phosphatase, ALP)试剂盒检测ALP活性;qRT-PCR检测MGF及成骨分化基因ALP、Runt相关转录因子2（runt-related transcription factor 2,Runx2）和骨桥蛋白（osteopontin,OPN）的转录水平,Western 免疫印迹检测对MEK/EKR1/2信号通路的影响。采用SPSS19.0软件包对结果进行统计学分析。结果:CS呈时间依赖性促进hPDLCs中MGF的表达（P<0.05）。与对照组相比,转染si-MGF显著抑制细胞中MGF的表达（P<0.05）,沉默MGF显著抑制CS诱导的hPDLCs中ALP的活性（P<0.05）及成骨分化相关基因ALP、Runx2及OPN的转录（均P<0.05）。与CS处理组相比,沉默MGF后,CS诱导的MMP-1及MMP-2的水平显著降低（P<0.05）;刺激MGF则进一步加强CS诱导的成骨分化及MMP-1、MMP-2的表达。沉默MGF可抑制CS激活的p-ERK的表达,而刺激MGF则进一步促进p-ERK的表达（P<0.05）。U0126预处理显著降低MGF加强的促成骨分化及MMP的表达（P<0.05）。结论:MGF通过MEK/ERK1/2信号通路的活化参与周期性牵张力作用下牙周膜细胞的成骨分化及MMP表达。

关键词: 口腔正畸, MGF, 人牙周膜细胞, 周期性牵张力, 成骨分化, MMPs

Abstract: PURPOSE: To explore the role and mechanism of mechano-growth factor (MGF) in cyclic stretch (CS)-induced osteogenic differentiation and MMP-1, MMP-2 expression in human periodontal ligament cells (hPDLCs). METHODS: HPDLCs were isolated and transfected with si-MGF, or stimulated with MGF or MEK/ERK pathway inhibitor U0126. Cells were cultured in Flexercell system with 10% elongation at 0.1 Hz. An alkaline phosphatase (ALP) kit was used to detect ALP activity. QRT-PCR assay was performed to determine the transcript levels of MGF and osteogenic genes, including ALP, runt-related transcription factor 2 (Runx2) and osteopontin (OPN). Western bot was used to evaluate the effect on MEK/EKR1/2 signaling. Statistical analysis was performed using SPSS 19.0 software package. RESULTS: CS induced the expression of MGF in hPDLCs in a time-dependent manner (P<0.05). In contrast to the control group, transfection with si-MGF inhibited the expression of MGF in hPDLCs (P<0.05). Moreover, cessation of MGF dramatically suppressed ALP activity (P<0.05) and the expression of osteogenic gene ALP, Runx2 and OPN (P<0.05) in hPDLCs. Furthermore, down-regulation of MGF restrained the expression of MMP-1 and MMP-2, in contrast to CS group (P<0.05). Conversely, stimulation with MGF further enhanced the effects of CS on osteogenic differentiation of hPDLCs and MMP-1, MMP-2 expression (P<0.05). Additionally, MGC silencing abrogated CS-induced expression of p-ERK (P<0.05), which was further enhanced following MGF treatment (P<0.05). Simultaneously, precondition with U0126 antagonized MGF-enhanced effects on CS-triggered osteogenic differentiation and MMP-1, MMP-2 expression (P<0.05). CONCLUSIONS: Mechano-growth factor regulates cyclic stretch-induced osteogenic differentiation and MMP-1, MMP-2 expression in human periodontal ligament cells by activating MEK/ERK1/2 signaling pathway.

Key words: Orthodontics, MGF, Human periodontal ligament cells, Cyclic stretch, Osteogenic differentiation, MMPs

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