肌球蛋白10对骨吸收过程中破骨细胞分化及功能的影响  

Effect of myosin 10 on osteoclast differentiation and function during bone resorption

作　　者：邓晔坤[1] 单冰晨[1] 周震涛[1] 张鹏[1] 周晓中[1] Deng Yekun;Shan Bingchen;Zhou Zhentao;Zhang Peng;Zhou Xiaozhong(Department of Orthopedics,the Second Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]苏州大学附属第二医院骨科,215000

出　　处：《中华实验外科杂志》2020年第6期1056-1058,共3页Chinese Journal of Experimental Surgery

基　　金：苏州市"科教兴卫"青年科技项目(KJXW2017011)。

摘　　要：目的观察肌球蛋白10(Myo 10)对破骨细胞的分化及功能的影响,并探讨其机制。方法采用核因子-κB(NF-κB)受体活化因子配体(RANKL)和巨噬细胞集落生长因子(M-CSF)诱导RAW264.7细胞(中国科学院上海细胞生物研究所)向破骨细胞分化,随机分为抗Myo 10组和对照组。应用抗酒石酸酸性磷酸酶(TRAP)染色法鉴定并定量TRAP+破骨样细胞的数量。使用反转录-聚合酶链反应(RT-PCR)技术检测破骨细胞分化及功能特异性基因的转录水平变化。将C57BL/6小鼠随机分为Myo 10 KO组和对照组,通过组织学染色苏木精-伊红(HE)和MicroCT评估Myo 10抑制对骨量的影响。两组间比较采用t检验。结果TRAP染色显示Myo 10抑制将促进破骨细胞的分化效率,TRAP+破骨细胞的数量在抗Myo 10组为(65.6±8.4)个,高于对照组的(41.3±6.2)个,差异有统计学意义(t=4.013,P<0.05);RT-PCR结果提示Myo 10抑制反而提高了基质金属蛋白酶-9(MMP-9)、活化T-细胞核因子1(NFATc1)及组织蛋白酶K(CtsK)的表达,分别为对照组表达值的(2.2±0.3)、(1.9±0.2)和(1.5±0.2)倍,差异有统计学意义(t=5.410、6.086、3.602,P<0.05);在体实验则提示Myo 10抑制后将增强骨吸收功能,出现骨质疏松样改变。结论Myo 10的抑制能促进NFATc1的表达,促进破骨细胞分化和骨吸收功能,导致骨量减少。Objective To study the effect of myosin 10(Myo 10)on the differentiation and function of osteoclasts(OC),and to explore its potential mechanism.Methods The differentiation of RAW264.7 cells into osteoclasts was induced by receptor activator for nuclear factor kappa B ligand(RANKL)and macrophage colony-stimulating factor(M-CSF).The cells were randomly divided into anti-Myo 10 group and control group.The number of tartrate resistant acid phosphatase(TRAP)+osteoclasts was identified and quantified by TRAP staining.Reverse transcriptase-polymerase chain reaction(RT-PCR)was used to detect osteoclast differentiation and functional specific gene transcription.C57BL/6 mice were randomly divided into two groups:Myo 10 KO group and control group.Results TRAP staining showed that Myo 10 inhibition could promote the differentiation efficiency of osteoclasts.The number of TRAP+osteoclasts in anti-Myo 10 group was(65.6±8.4),significantly greater than that in control group(41.3±6.2)(t=4.013,P<0.05).RT-PCR results suggested that Myo 10 inhibition instead increased the expression levels of matrix metalloproteinase-9(MMP-9),activated T-cell nuclear factor 1(NFATc1)and cathepsin K(CtsK),which were(2.2±0.3),(1.9±0.2)and(1.5±0.2)times of the expression value in the control group,respectively(t=5.410,6.086,3.602,P<0.05).The in vivo experiments suggested that Myo 10 inhibition could enhance bone resorption function and cause osteoporosis-like changes.Conclusion The inhibition of Myo 10 can promote the expression of NFATc1,osteoclast differentiation and bone resorption,and decrease bone mass.

关 键 词：Myo 10 破骨细胞 小干扰RNA MICROCT 骨吸收 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]