国际口腔医学杂志 ›› 2017, Vol. 44 ›› Issue (6): 679-685.doi: 10.7518/gjkq.2017.06.011

• 论著 • 上一篇    下一篇

机械牵张力促进小鼠骨髓间充质干细胞的成骨向分化

薛令法1, 张岱尊2, 肖文林1, 于保军1   

  1. 1.青岛大学附属医院口腔颌面外科;山东省教育厅口腔重点实验室 青岛 266555;
    2.青岛大学附属医院儿童口腔科 青岛 266003
  • 收稿日期:2016-12-23 修回日期:2017-07-20 出版日期:2017-11-01 发布日期:2017-11-01
  • 通讯作者: 肖文林,副教授,博士,Email:wenlinxiao@sina.com
  • 作者简介:薛令法,主治医师,硕士,Email:xuelingfa@163.com
  • 基金资助:
    山东省高校科技计划(J12LK57); 青岛市黄岛区应用研究与公共卫生专项基金(2014-1-84)

Mechanical strain induces mouse bone mesenchymal stem cells osteogenic differentiation

Xue Lingfa1, Zhang Daizun2, Xiao Wenlin1, Yu Baojun1   

  1. 1. Dept. of Oral and Maxillofacial Surgery, The Affiliated Hospital of Qingdao University; The Oral Key Laboratory of Education Department of Shandong Province, Qingdao 266555, China;
    2. Dept. of Pediatric Dentistry, The Affiliated Hospital of Qingdao University, Qingdao 266003, China
  • Received:2016-12-23 Revised:2017-07-20 Online:2017-11-01 Published:2017-11-01
  • Supported by:
    ; This study supported by Science and Technology Program in Colleges and Universities of Shandong(J12LK57) and Special Fund for Applied Study and Public Health, Huangdao District, Qingdao(2014-1-84).

摘要: 目的 旨在研究p38分裂原激活的蛋白激酶(MAPK)信号通路和转录因子Osterix在间断性机械牵张力刺激下促进小鼠骨髓间充质干细胞(BMSC)成骨向分化的作用机制。方法 将C57BL/6J小鼠BMSC分为空白对照组、牵张力组和牵张力阻断组(p38MAPK通路抑制剂SB203580+牵张力),采用Flexercell体外细胞力学加载装置,施加频率为0.5 Hz、形变率为0.8%的牵张力,每天2次,每次30 min,分别在实验第1、3、5 d收获BMSC。实时荧光定量聚合酶链反应(RT-PCR)检测成骨基因ALPCOL IOCN的mRNA水平变化情况,Western blot检测P-p38-MAPK蛋白的表达情况。通过小干扰核糖核酸(siRNA)技术沉默小鼠BMSC的osterix基因,Western blot检测Osterix 蛋白的表达情况,RT-PCR检测成骨相关基因ALPCOL IOCN的mRNA水平变化情况。结果 牵张力作用后,可观察到成骨相关基因ALPCOL IOCN及转录因子Osterix的mRNA水平增高。沉默osterix后,小鼠BMSC成骨相关基因ALPCOL IOCN的mRNA水平也随之降低。Western blot结果显示,牵张力组小鼠BMSC中Osterix和P- p38-MAPK的蛋白质水平明显高于对照组(P<0.05)。SB203580作用后,小鼠BMSC成骨相关基因ALPCOL IOCNosterix的mRNA水平降低。结论 间断性牵张力可通过活化p38MAPK-Osterix通路促进小鼠BMSC成骨分化。

关键词: 间断性牵张力, 骨髓间充质干细胞, 成骨向分化, p38MAPK信号通路

Abstract: Objective This study aimed to investigate the stimulating mechanism of p38 mitogen-activated protein kinase(MAPK) signaling pathway and Osterix under intermittent mechanical stretch tension to accelerate the osteogenic differentiation of bone mesenchymal stem cells(BMSCs) in mice. Methods Three groups of C57BL/6J BMSC, namely, blank control group, strain group, and inhibitor group(p38MAPK signal pathway inhibitor SB203580 + strain), were prepared and exposed to 0.8% intermittent mechanical strain at 0.5 Hz twice a day for 30 min at each time by using a Flexercell strain unit. The cells in the three groups were then harvested on days 1, 3, and 5, respectively. Changes in the mRNA expression of ALP, COL, and OCN were detected through real time-polymerase chain reaction(RT-PCR) and the protein expression of P-p38MAPK was observed through Western blot analysis. osterix gene was knocked down by small interfering RNA(siRNA), and Osterix protein expression was determined through Western blot. The mRNA expression levels of ALP, COL I, and OCN were identified through RT-PCR. Results Mechanical tension force could promote the mRNA expression of ALP, COL I, OCN, and osterix. osterix silencing could decrease the mRNA expression of ALP, COL I, and OCN. Western blot revealed that the protein expression levels of Osterix and P-p38MAPK in the strain group were significantly higher than those in the control group(P<0.05). The mRNA expression of ALP, COL I, OCN, and osterix decreased after SB203580 was used. Conclusion Intermittent mechanical strain promotes the osteogenic differentiation of BMSC via the p38MAPK-Osterix pathways.

Key words: intermittent stretching force, bone marrow stem cell, osteoblastic differentiation, p38MAPK signal pathway

中图分类号: 

  • Q786
[1] Koike M, Shimokawa H, Kanno Z, et al. Effects of mechanical strain on proliferation and differentiation of bone marrow stromal cell line ST2[J]. J Bone Miner Metab, 2005, 23(3):219-225.
[2] Ming LG, Ge BF, Wang MG, et al. Comparison between 8-prenylnarigenin and narigenin concerning their activities on promotion of rat bone marrow stromal cells’ osteogenic differentiation in vitro [J]. Cell Prolif, 2012, 45(6):508-515.
[3] Chen D, Li Y, Zhou Z, et al. Synergistic inhibition of Wnt pathway by HIF-1α and osteoblast-specific transcription factor osterix(Osx) in osteoblasts[J]. PLoS One, 2012, 7(12):e52948.
[4] Zhang C, Li J, Zhang L, et al. Effects of mechanical vibration on proliferation and osteogenic differentia-tion of human periodontal ligament stem cells[J]. Arch Oral Biol, 2012, 57(10):1395-1407.
[5] Zhao Y, Wang C, Li S, et al. Expression of Osterix in mechanical stress-induced osteogenic differentia-tion of periodontal ligament cells in vitro [J]. Eur J Oral Sci, 2008, 116(3):199-206.
[6] Kang KS, Lee SJ, Lee H, et al. Effects of combined mechanical stimulation on the proliferation and differentiation of pre-osteoblasts[J]. Exp Mol Med, 2011, 43(6):367-373.
[7] Soleimani M, Nadri S. A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow[J]. Nat Protoc, 2009, 4(1):102-106.
[8] Luther F, Layton S, McDonald F. Orthodontics for treating temporomandibular joint(TMJ) disorders[J]. Cochrane Database Syst Rev, 2010, 7(7):CD006541.
[9] Ellegaard M, Kringelbach T, Syberg S, et al. The effect of PTH(1-34) on fracture healing during dif-ferent loading conditions[J]. J Bone Miner Res, 2013, 28(10):2145-2155.
[10] Rachmiel A, Emodi O, Gutmacher Z, et al. Oral and dental restoration of wide alveolar cleft using dis-traction osteogenesis and temporary anchorage de-vices[J]. J Craniomaxillofac Surg, 2013, 41(8):728- 734.
[11] Suzuki A, Guicheux J, Palmer G, et al. Evidence for a role of p38 MAP kinase in expression of alkaline phosphatase during osteoblastic cell differentiation [J]. Bone, 2002, 30(1):91-98.
[12] Xiao G, Jiang D, Thomas P, et al. MAPK pathways activate and phosphorylate the osteoblast-specific transcription factor, Cbfa1[J]. J Biol Chem, 2000, 275(6):4453-4459.
[13] Coulthard LR, White DE, Jones DL, et al. p38 MAPK : stress responses from molecular mechanisms to therapeutics[J]. Trends Mol Med, 2009, 15(8):369- 379.
[14] Zhao Y, Song T, Wang W, et al. P38 and ERK1/2 MAPKs act in opposition to regulate BMP9-induced osteogenic differentiation of mesenchymal proge-nitor cells[J]. PLoS One, 2012, 7(8):e43383.
[15] Wang L, Li JY, Zhang XZ, et al. Involvement of p38MAPK/NF-κB signaling pathways in osteoblasts differentiation in response to mechanical stretch[J]. Ann Biomed Eng, 2012, 40(9):1884-1894.
[16] Du QC, Zhang DZ, Chen XJ, et al. The effect of p38MAPK on cyclic stretch in human facial hyper-trophic scar fibroblast differentiation[J]. PLoS One, 2013, 8(10):e75635.
[17] Simmons CA, Matlis S, Thornton AJ, et al. Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase(ERK1/2) signaling pathway [J]. J Biomech, 2003, 36(8):1087-1096.
[18] Zhang P, Wu Y, Dai Q, et al. p38-MAPK signaling pathway is not involved in osteogenic differentiation during early response of mesenchymal stem cells to continuous mechanical strain[J]. Mol Cell Biochem, 2013, 378(1/2):19-28.
[19] Burr DB, Milgrom C, Fyhrie D, et al. In vivo mea-surement of human tibial strains during vigorous activity[J]. Bone, 1996, 18(5):405-410.
[20] Qi MC, Zou SJ, Han LC, et al. Expression of bone-related genes in bone marrow MSCs after cyclic mechanical strain: implications for distraction osteogenesis[J]. Int J Oral Sci, 2009, 1(3):143-150.
[1] 张静怡,李丹薇,孙宇,雷雅燕,刘涛,龚瑜. 复合树脂及复合体对成骨细胞毒性及成骨向分化的影响[J]. 国际口腔医学杂志, 2022, 49(4): 412-419.
[2] 陈野, 周丰, 邬琼辉, 车会凌, 李佳璇, 申佳琪, 罗恩. 脂联素对骨髓间充质干细胞的作用及其调控机制[J]. 国际口腔医学杂志, 2021, 48(1): 58-63.
[3] 杨叶青,陈明,吴补领. 环状非编码RNA在间充质干细胞成骨向分化中作用的研究进展[J]. 国际口腔医学杂志, 2020, 47(3): 257-262.
[4] 刘俊圻,陈艺尹,杨文宾. RNA腺嘌呤6-甲基化修饰调控骨髓间充质干细胞成骨向分化的研究进展[J]. 国际口腔医学杂志, 2020, 47(3): 263-269.
[5] 王润婷,房付春. 非编码RNA调控人牙周膜干细胞成骨向分化的研究进展[J]. 国际口腔医学杂志, 2020, 47(2): 138-145.
[6] 张建康, 卫俊俊, 唐曌隆, 余云波, 敬伟. Wnt和Notch通路在老龄个体骨髓间充质干细胞成骨中的调控[J]. 国际口腔医学杂志, 2017, 44(4): 459-465.
[7] 黄奕华 凌均棨. Toll样受体2和4在细胞成骨向分化中的作用[J]. 国际口腔医学杂志, 2015, 42(4): 492-495.
[8] 王涛1 廖天安1 王鸿1 邓伟1 于大海2. 血管内皮生长因子基因修饰骨髓间充质干细胞移植于放射治疗后组织的实验研究[J]. 国际口腔医学杂志, 2014, 41(2): 133-136.
[9] 苟文亭综述 叶玲 谭红审校. 牙髓干细胞分化的研究进展[J]. 国际口腔医学杂志, 2012, 39(3): 380-383.
[10] 王婷婷,万乾炳,. 骨髓干细胞应用于牙槽骨修复的展望[J]. 国际口腔医学杂志, 2008, 35(S1): -.
[11] 谢谦 黄洪章. 骨髓间充质干细胞与骨相关研究[J]. 国际口腔医学杂志, 2004, 31(03): 181-182.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] 王昆润. 二甲亚砜和双氯芬酸并用治疗根尖周炎[J]. 国际口腔医学杂志, 1999, 26(06): .
[2] 汤庆奋,王学侠. 17β-雌二醇对人类阴道和口腔颊粘膜的渗透性[J]. 国际口腔医学杂志, 1999, 26(06): .
[3] 潘劲松. 颈总动脉指压和颈内动脉球囊阻断试验在大脑血液动力学中的不同影响[J]. 国际口腔医学杂志, 1999, 26(05): .
[4] 王昆润. 后牙冠根斜形牙折的治疗[J]. 国际口腔医学杂志, 1999, 26(05): .
[5] 杨锦波. 嵌合体防龋疫苗的研究进展[J]. 国际口腔医学杂志, 1999, 26(05): .
[6] 王昆润. 下颔骨成形术用网状钛板固定植骨块[J]. 国际口腔医学杂志, 1999, 26(04): .
[7] 汪月月,郭莉莉. 口腔机能与老化—痴呆危险因素流行病学研究[J]. 国际口腔医学杂志, 1999, 26(04): .
[8] 丁刚. 应用硬组织代用品种植体行丰颏术[J]. 国际口腔医学杂志, 1999, 26(04): .
[9] 田磊. 局部应用脂多糖后结合上皮反应性增生的变化[J]. 国际口腔医学杂志, 1999, 26(04): .
[10] 戴青. 口腔念珠菌病的新分类[J]. 国际口腔医学杂志, 1999, 26(04): .