低强度高频率振动对成骨细胞生物学特性的影响

查丁胜; 邓轩赓; 陈建庭

doi:10.13418/j.issn.1001-165x.2018.02.010

中国临床解剖学杂志 ›› 2018, Vol. 36 ›› Issue (2) : 165-173. DOI: 10.13418/j.issn.1001-165x.2018.02.010

实验研究

低强度高频率振动对成骨细胞生物学特性的影响

查丁胜¹ ，　邓轩赓² ，　陈建庭³

作者信息 +

Effects of low magtitude and high frequency vibration on the biological characteristics of osteoblasts

ZHA Ding-sheng¹, DENG Xuan-geng², CHEN Jian-ting³

Author information +

文章历史 +

摘要

目的　研究低强度高频率振动（low-magnitude high-frequency vibration, LMHFV）对成骨细胞生物学特性的影响。方法　建立LMHFV加载MC3T3-E1细胞模型，观察不同频率LMHFV对MC3T3-E1细胞OPG/RANKL浓度比的影响，获得OPG/RANKL浓度比最高的频率（F）为后续研究频率；以0 Hz为对照，观察LMHFV对MC3T3-E1细胞碱性磷酸酶 (ALP)、骨钙素(OCN) mRNA和蛋白活性，及钙化结节形成的影响；LMHFV加载形成的条件培养液（CMF）孵育RAW264.7细胞，观察CMF对破骨细胞抗酒石酸酸性磷酸酶（TRAP）染色、多核破骨细胞形成、TRAP mRNA及蛋白活性的影响；观察LMHFV对MC3T3-E1细胞环氧化酶2(COX-2)蛋白水平的表达及COX-2抑制剂NS-398对LMHFV影响MC3T3-E1细胞分化的作用。结果　30 Hz LMHFV获得OPG/RANKL浓度比最高，促进ALP、OCN mRNA及蛋白活性增加，增加钙化结节形成。30 Hz LMHFV形成的CM抑制RAW264.7细胞向多核破骨细胞分化，抑制TRAP mRNA及活性；LMHFV可诱导COX-2蛋白水平增加，NS-398能抑制LMHFV促进成骨细胞分化。结论　30 Hz的LMHFV对MC3T3-E1细胞OPG/RANKL浓度比及成骨分化具有积极的影响，通过调控成骨细胞OPG/RANKL浓度比间接抑制骨吸收，COX-2通路参与了LMHFV对成骨细胞生物学特性的调节作用。

Abstract

Objective　To study the effects of low-magnitude high-frequency vibration (LMHFV) on the biological characteristics of osteoblasts.　Methods　To establish a model of MC3T3-E1 cells loaded with LMHFV, and observe the effects of LMHFV at different frequencies on the concentration ratio of OPG / RANKL in MC3T3-E1 cells, the frequency with the highest OPG / RANKL concentration ratio (F) was the frequency of follow-up studies. With 0 Hz as a control, to observe the effects of LMHFV on the expression of akialine phosphatase (ALP), osteocalcin (OCN) mRNA and protein activity, and calcified nodules in MC3T3-E1 cells; RAW264.7 cells were incubated with conditioned medium (CMF) loaded with LMHFV, and to observe the effect of CMF on TRAP staining, multinucleated osteoclast formation, TRAP mRNA and protein activity in osteoclasts; to study the expression of cyclooxygenase-2 (COX-2) protein level in MC3T3-E1 cells loaded with LMHFV and the effect of COX-2 inhibitor NS-398 on the differentiation of MC3T3-E1 cells induced by LMHFV.　Results　The highest concentration ratio of OPG / RANKL was obtained at 30 Hz in LMHFV, which promoted the increase of ALP, OCN mRNA and protein activity and increased the formation of calcified nodules. CM formed by LMHFV at 30 Hz inhibited the differentiation of RAW264.7 cells into multinucleated osteoclasts and inhibited the TRAP mRNA and protein activity. LMHFV induced an increase in COX-2 protein level and NS-398 inhibited LMHFV-induced osteoblast differentiation.　Conclusion　LMHFV at 30 Hz had a positive effect on OPG / RANKL concentration ratio and osteogenic differentiation in MC3T3-E1 cells, indirectly suppresses bone resorption by regulating the osteoblast OPG / RANKL concentration ratio, the COX-2 pathway is involved in the regulation of osteoblast biological properties by LMHFV.

导出引用

查丁胜,邓轩赓,陈建庭. 低强度高频率振动对成骨细胞生物学特性的影响[J]. 中国临床解剖学杂志. 2018, 36(2): 165-173 https://doi.org/10.13418/j.issn.1001-165x.2018.02.010

ZHA Ding-sheng, DENG Xuan-geng, CHEN Jian-ting. Effects of low magtitude and high frequency vibration on the biological characteristics of osteoblasts[J]. Chinese Journal of Clinical Anatomy. 2018, 36(2): 165-173 https://doi.org/10.13418/j.issn.1001-165x.2018.02.010

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