运动上调GPR48-RANKL通路影响 2型糖尿病小鼠破骨细胞分化

doi:10.15942/j.jcsu.2017.03.014

成都体育学院学报 ›› 2017, Vol. 43 ›› Issue (3): 90-95.doi: 10.15942/j.jcsu.2017.03.014

运动上调GPR48-RANKL通路影响 2型糖尿病小鼠破骨细胞分化

陈祥和¹, 孙朋², 杨念恩³, 李世昌², 徐会金², 张娜娜⁴

1. 扬州大学体育学院,江苏扬州 225127;
2. 华东师范大学体育与健康学院,上海 200241;
3. 安庆师范大学体育学院,安徽安庆 246000;
4. 海口经济学院拳星时代体育学院,海南海口 571127

收稿日期:2017-01-19 出版日期:2017-05-25 发布日期:2017-05-25
作者简介:陈祥和,博士,讲师,研究方向:运动与骨代谢的机制研究,E-mail:huashixh@163.com
基金资助:
江苏省教育科学“十三五”规划课题“跳跃运动对小学生体质与骨健康的影响及其相关性分析研究”(T-b/2016/08);中国博士后科学基金第8批特别资助课题“运动强度调控肥胖青少年生理血压的自主神经机制的研究”(2015T80412)。

Effects of Exercise on Osteoclast Differentiation through the Up-regulation GPR48-RANKL Pathway in Type 2 Diabetic Mice

CHEN Xianghe¹, SUN Peng², YANG Nianen³, LI Shichang², XU Huijin², ZHANG Nana²

1. College of Physical Education, Yangzhou University, Yangzhou Zhejiang 225127;
2. College of Physical Education and Health, East China Normal University, Shanghai 200241;
3. College of Physical Education, AnQing Normal University, Anqing Anhui 24600;
4. QUANXING College of Physical, Haikou College of Economics, Haikou Hainan 571127

Received:2017-01-19 Online:2017-05-25 Published:2017-05-25

摘要/Abstract

摘要： 目的: 探究T2DM小鼠OC分化变化及不同力学刺激通过GPR48-RANKL通路对T2DM小鼠OC分化的分子调控作用。方法: 40只4周龄雄性C57BL/6小鼠,适应性喂养1周后随机分为T2DM造模组(30只)和正常对照组(ZC, 10只)。T2DM造模组小鼠6周高脂膳食结束空腹12 h后注射STZ,2周后检测小鼠血糖,有27只造模成功并将其随机分为T2DM对照组(TC, 9只)、T2DM游泳组(TS, 9只)和T2DM游泳组(TD, 9只),TS和TD组小鼠分别进行8周游泳和下坡跑训练。结束后,取右侧股骨并利用RT-PCR法检测相关因子mRNA表达;取左侧胫骨并利用West-blotting法检测相关因子蛋白表达;取小鼠BMM并诱导其向OC分化,利用TRAP染液对OC染色;取右侧胫骨并利用游标卡尺检测其大小。结果: 与ZC组相比,TC组GPR48、OPG、RANKL、RANK、NFATc2、CTSK的mRNA和GPR48、RANKL、RANK蛋白表达均显著变化(P<0.05或P<0.01),OC数量显著增多,远端冠状面宽度和近端冠状面宽度显著变小(P<0.05)。与TC组相比,TS组GPR48、OPG、RANKL、NFATc2、CTSKmRNA和GPR48、RANKL蛋白表达均显著变化(P<0.05或P<0.01),OC数量显著减少;TD组GPR48、OPG、RANKL、RANK、NFATc2、CTSKmRNA和GPR48、RANKL、RANK蛋白表达均显著上调(P<0.05或P<0.01),OC数量显著减少,胫骨长度和中间矢状轴宽度显著减少(P<0.05或P<0.01)。与TS组相比,TD组OPG和CTSKmRNA及GPR48、RANK、RANKL蛋白表达均显著变化(P<0.05或P<0.01),OC数量显著减少。结论: T2DM小鼠OC分化显著增强;直接作用力激活T2DM小鼠骨中GPR48-RANKL通路,进而抑制RANK及其下游靶基因表达,抑制OC分化,且其作用效果优于间接作用力。

关键词: G蛋白偶联受体48, 核因子κB受体活化因子配体, 不同力学刺激, Ⅱ型糖尿病, 破骨细胞

Abstract: Objective: :To investigate the change of OC differentiation in T2DM mice and effect of different mechanical stimulation on OC differentiation by GPR48- RANKL pathway in T2DM mice.Methods: Forty four-week old C57BL/6 male mice were randomly divided into normal control group (ZC) and T2DM group. Use the method of high-fat diet and STZ to build the model of T2DM mice, and the T2DM mice were randomly divided into T2DM control group (TC), T2DM swimming group (TS) and T2DM downhill run group (TD). And use the swimming and downhill running to train the T2DM mice for eight weeks. After this, use the RT-PCR to test the mRNA expression in the left tibia. Use the WB to test the protein expression in right femur. Take the BMM and induced it differentiation into osteoclast, use the TRAP solution to dye the osteoclasts. Use the vernier caliper to test the shape size of right tibia.Results: Compared to ZC group, the mRNA expression of GPR48、OPG、RANKL、RANK, NFATc2, CTSK and protein expression of GPR48、RANKL、RANK of TC group were all significantly changed (P<0.05 or P<0.01), the number of OC was significantly increased, the width of distal coronary surface and the surface of the proximal coronary was significantly reduced(P<0.05). Compared to TC group, the mRNA expression of GPR48, OPG, RANKL, NFATc2, CTSK and the protein expression of GPR48 and RANKLof TS group were significantly changed(P<0.05 or P<0.01). The number of osteoclast was significantly reduced. The mRNA expression of GPR48, OPG, RANKL, RANK, NFATc2, CTSK and the protein expression of GPR48, RANKL, RANK were all significantly changed(P<0.05 or P<0.01). The number of osteoclast was significantly reduced, the length of tibia and width of the middle sagittal axis were significantly increased(P<0.05). Compared to TS group, the mRNA expression of OPG, CTSK and the protein expression of GPR48, RANK, RANKL of TD group were significantly changed(P<0.05 or P<0.01), OC number were drastically reduced.Conclusion: The osteoclast differentiation of type 2 diabetic mice was significantly increased. The direct mechanical stimulation activited the GPR48-RANKL pathway, which inhibited the RANK and their target gene expression, and then inhibited the osteoclast differentiation of type 2 diabetic mice. The effect of direct mechanical stimulation was better than that of indirect one.

Key words: G Protein Coupled Receptor 48, Receptor Activator for Nuclear Factor-κ B Ligand, Different Mechanical Stimulation, Type 2 Diabetes Mellitus, Osteoclast

中图分类号:

G804.2

陈祥和, 孙朋, 杨念恩, 李世昌, 徐会金, 张娜娜. 运动上调GPR48-RANKL通路影响 2型糖尿病小鼠破骨细胞分化[J]. 成都体育学院学报, 2017, 43(3): 90-95.

CHEN Xianghe¹, SUN Peng², YANG Nianen³, LI Shichang², XU Huijin², ZHANG Nana². Effects of Exercise on Osteoclast Differentiation through the Up-regulation GPR48-RANKL Pathway in Type 2 Diabetic Mice[J]. Journal of Chengdu Sport University, 2017, 43(3): 90-95.

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运动上调GPR48-RANKL通路影响 2型糖尿病小鼠破骨细胞分化

Effects of Exercise on Osteoclast Differentiation through the Up-regulation GPR48-RANKL Pathway in Type 2 Diabetic Mice

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