周期性牵张力介导BMP9通过PI3K/AKT信号通路调控人牙周膜成纤维细胞成骨分化

doi:10.13418/j.issn.1001-165x.2019.06.010

中国临床解剖学杂志 ›› 2019, Vol. 37 ›› Issue (6) : 656-661. DOI: 10.13418/j.issn.1001-165x.2019.06.010

张超¹，　胡静²，　张君怡³，　魏克元⁴，　孙海豹⁵，　黄克强¹

作者信息 +

Cyclic tension force-mediated BMP9 regulates osteogenic differentiation of human periodontal ligament fibroblasts via PI3K/AKT signaling pathway

ZHANG Chao，HU Jing，ZHANG Jun-yi，WEI Ke-yuan，SUN Hai-bao，HUANG Ke-qiang

Author information +

文章历史 +

摘要

目的　研究在周期性牵张力介导下，骨形态发生蛋白9（bone morphogenetic proteins 9，BMP9）能否激活PI3K/AKT信号通路调控人牙周膜成纤维细胞（human periodontal ligament fibroblasts，hPDLFs）成骨分化。方法　利用FlexCell系统对体外培养的hPDLFs加载正弦波、形变率10%、频率0.5 Hz的周期性牵张力，免疫荧光法检测细胞骨架蛋白的表达及分布，qPCR检测RUNX2、OCN、OPN、OSX mRNA的表达情况，免疫印迹法检测成骨标志蛋白OCN、OPN的表达，加入PI3K信号抑制剂LY294002后AKT、P-AKT以及OCN、OPN的变化。结果　与对照组比较，6 h、12 h组的细胞骨架蛋白表达增多且呈受力方向分布，OCN、OPN表达上调，差异有统计学意义（P<0.05），qPCR检测mRNA的表达趋势与蛋白检测结果一致。结论　周期性牵张力介导BMP9可以通过PI3K/AKT信号通路调控hPDLFs成骨分化。

Abstract

Objective　To investigate whether bone morphogenetic proteins 9 (BMP9) can activate PI3K/AKT signaling pathway to regulate osteogenic differentiation of human periodontal ligament fibroblasts (hPDLFs) under cyclic tension force.　Methods　The cultured in vitro hPDLFS were loaded with sine wave, deformation rate 10%, frequency 0.5 Hz periodic tension by FlexCell system. Distribution and changes of cytoskeletal protein expression were observed by immunofluorescence method. Changes of mRNA expression of RUNX2, OCN, OPN and OSX were observed by qPCR. Cytoskeletal protein OCN, OPN expressions and changes of AKT, P-AKT, OCN, OPN after adding P13K signal inhibitor LY294002 were detected by Western blot.　Results　Compared with control group, there were a significant increase of the expression of cytoskeletion-associated protein, up-regulation of the expression of osteogenic markers in 6 h and 12 h groups (P<0.05). The trend of mRNA expression was consistent with the result of protein detection.　Conclusions　Cyclic tension force-mediated BMP9 can regulate osteogenic differentiation of hPDLFs through PI3K/AKT signaling pathway.

导出引用

张超, 胡静, 张君怡, 魏克元, 孙海豹, 黄克强. 周期性牵张力介导BMP9通过PI3K/AKT信号通路调控人牙周膜成纤维细胞成骨分化[J]. 中国临床解剖学杂志. 2019, 37(6): 656-661 https://doi.org/10.13418/j.issn.1001-165x.2019.06.010

ZHANG Chao, HU Jing, ZHANG Jun-yi, WEI Ke-yuan, SUN Hai-bao, HUANG Ke-qiang. Cyclic tension force-mediated BMP9 regulates osteogenic differentiation of human periodontal ligament fibroblasts via PI3K/AKT signaling pathway[J]. Chinese Journal of Clinical Anatomy. 2019, 37(6): 656-661 https://doi.org/10.13418/j.issn.1001-165x.2019.06.010

中图分类号： R783.5

参考文献

[1] Nettelhoff L, Grimm S, Jacobs C, et al. Influence of mechanical compression on human periodontal ligament fibroblasts and osteoblasts[J]. Clin Oral Investig, 2016, 20(3):621-629.
[2] Jacobs C, Walter C, Ziebart T, et al. Mechanical loading influences the effects of bisphosphonates on human periodontal ligament fibroblasts[J]. Clin Oral Investig, 2015, 19(3):699-708.
[3] Pirjamalineisiani A, Sarafbidabad M, Jamshidi N, et al. Finite element analysis of post dental implant fixation in drilled mandible sites[J]. Comput Biol Med, 2016, 81(6):159-166.
[4] Kang Q, Sun MH, Cheng H, et al. Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery[J]. Gene Ther, 2004, 11(17):1312-1320.
[5] Luther G, Wagner ER, Zhu G, et al. BMP-9 induced osteogenic differentiation of mesenchymal stem cells: molecular mechanism and therapeutic potential[J]. Curr Gene Ther, 2011, 11(3): 229-240.
[6] Cheng H, Jiang W, Phillips FM, et al. Osteogenic activity of the fourteen types of human bone morphogenetic proteins (BMPs)[J]. J Bone Joint Surg Am, 2003, 85(8): 1544-1552.
[7] Peng Y, Kang Q, Luo Q, et al. Inhibitor of DNA binding/differentiation helix-loop-helix proteins mediate bone morphogenetic protein-induced osteoblast differentiation of mesenchymal stem cells[J]. J Biol Chem, 2004, 279(31): 32941-32949.
[8] Chang M, Lin H, Fu H, et al. MicroRNA-195-5p regulates osteogenic differentiation of periodontal ligament cells under mechanical loading[J]. J Cell Physiol, 2017, 232(12): 3762-3774.
[9] Luo Q, Kang Q, Si W, et al. Connective tissue growth factor (CTGF) is regulated by Wnt and bone morphogenetic proteins signaling in osteoblast differentiation of mesenchymal stem cells[J]. J Biol Chem, 2004, 279(53): 55958-55968.
[10] 杨泉林, 孟朋民, 石彬, 等. 骶韧带成纤维细胞受力后细胞骨架的变化[J]. 中国妇幼保健, 2014, 29(23): 3814-3816.
[11]Ren D, Wei F, Hu L, et al. Phosphorylation of Runx2, induced by cyclic mechanical tension via ERK1/2 pathway, contributes to osteodifferentiation of human periodontal ligament fibroblasts[J]. J Cell Physiol, 2015, 230(10): 2426-2436.
[12] 姚依村, 梁伟国, 叶冬平, 等. 细胞骨架与力学信号传导[J]. 中国组织工程研究, 2014, 18(7): 1109-1111.
[13] 吴莉萍, 韦曦, 凌均棨, 等. 细胞骨架及相关蛋白在人牙周韧带细胞成骨分化过程中的表达模式[J]. 中华口腔医学研究杂志(电子版), 2008, 2(5): 8-12.
[14]戚孟春, 胡静, 韩立赤, 等. 成骨细胞在机械力刺激下细胞骨架及细胞形态改变的体外研究[J]. 中国口腔颌面外科杂志, 2004, 2(3): 181-184.
[15]Fuchigami S, Nakamura T, Furue K, et al. Recombinant human bone morphogenetic protein-9 potently induces osteogenic differentiation of human periodontal ligament fibroblasts[J]. Eur J Oral Sci, 2016, 124(2): 151-157.
[16]Zhu JH, Liao YP, Li FS, et al. Wnt11 promotes BMP9-induced osteogenic differentiation through BMPs/Smads and p38 MAPK in mesenchymal stem cells[J]. J Cell Biochem, 2018, 119(11): 9462-9473.
[17] Furue K, Sena K, Sakoda K, et al. Involvement of the phosphoinositide 3-kinase/Akt signaling pathway in bone morphogenetic protein 9-stimulated osteogenic differentiation and stromal cell-derived factor 1 production in human periodontal ligament fibroblasts[J]. Eur J Oral Sci, 2017, 125(2): 119-126.
[18] Hamzehzadeh L, Atkin SL, Majeed M, et al. The versatile role of curcumin in cancer prevention and treatment: a focus on PI3K/AKT pathway[J]. J Cell Physiol, 2018, 233(10): 6530-6537.