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

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

张超¹，　胡静²，　张君怡³，　魏克元⁴，　孙海豹⁵，　黄克强¹

1. 锦州医科大学附属第二医院；　2. 锦州医科大学生命科学研究院；　3. 锦州医科大学康复与运动医学院，辽宁锦州 121000；　4. 赤峰学院附属医院，内蒙古赤峰 024000；　5. 绍兴市口腔医院，浙江绍兴 312000

收稿日期:2018-12-29 出版日期:2019-11-25 发布日期:2019-12-02
通讯作者: 黄克强，教授，主任医师，硕士生导师，E-mail：hkq9@163.com
作者简介:张超（1986-），主治医师，硕士研究生，主要从事周期性牵张力对hPDLFs成骨分化的影响及机制研究，E-mail：yayizhangchao@126.com
基金资助:
锦州医科大学校长基金（AH2015002）

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

1. The Second Affiliated Hospital of Jinzhou Medical University; 2. Jinzhou Medical University Life Science Research Institute; 3. Jinzhou Medical University School of Rehabilitation and Sports, Jinzhou 121000, Liaoning Province; 4. Affiliated Hospital of Chifeng University, Chifeng 024000, Inner Mongolia; 5. Shaoxing Stomatological Hospital, Shaoxing 312000, Zhejiang Province, China

Received:2018-12-29 Online:2019-11-25 Published:2019-12-02

摘要/Abstract

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。

关键词: 骨形态发生蛋白9, 　周期性牵张力, 　人牙周膜成纤维细胞, 　PI3K/AKT

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.

Key words: BMP-9；　Cyclic tensile force, 　hPDLFs, 　PI3K/AKT

中图分类号:

R783.5

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.

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.

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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

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

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