不同运动对生长期大鼠Wnt/β-catenin信号及软骨内成骨的影响
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摘要
目的:探索不同运动方式对生长期大鼠Wnt/β-catenin信号通路相关分子表达及软骨内成骨的影响。方法:对生长期大鼠进行6周的不同方式的运动,利用三点弯曲测骨强度,Micro-CT对股骨微结构进行检测,利用RT-PCR、west-bloting检测细胞因子mRNA和蛋白表达。结果:与C组相比,D组胫骨、股骨围度湿重有非常显著性增加(P<0.01),且D组围度较R组有显著性增加(P<0.05),骨强度测试D组有非常显著性增加(P<0.01);CT检测发现,股骨皮质骨BMD只有D组有显著性增加(P<0.05);Wnt4mRNA的表达,R组有显著性上调(P<0.05),D组有非常显著性上调(P<0.01);Wnt8mRNA、Wnt9mRNA的表达,R组和D组有非常显著性上调(P<0.01);GSK3βmRNA的表达,只有D组有显著性上调(P<0.05);β-cateninmRNA的表,R组和D组都有非常显著性上调(P<0.01),与R组相比,D组也有显著性上调(P<0.05);Runx2mRNA的表达,R组有显著性上调(P<0.05),D组的有非常显著性上调(P<0.01);Wnt8蛋白表达,R组和D组都有非常显著性增加(P<0.01),D组较R组也有非常显著性增加(P<0.01);β-catenin、Runx2蛋白表达,R组有显著性增加(P<0.05),D组有非常显著性增加(P<0.01),其中D组Runx2较R组也有显著性增加(P<0.05)。结论:跳跃运动通过Wnt/β-catenin信号通路,促进生长期大鼠生长板软骨内成骨,提高骨健康水平。
Objective:To explore the effects of different exercise modes on the expression of Wnt/β-catenin signaling pathway-related molecules and endochondral osteogenesis in growing rats. Methods:Growing rats were exercised in different ways for 6 weeks. Bone strength was measured by three-point bending,femoral micro-structure was detected by micro-CT,and cytokine gene and protein expression were detected by RT-PCR and western-bloting. Results:Compared with group C,the wet weight of tibia and femur circumference in group D increased significantly(P<0.01),and group D increased significantly(P<0.05). Bone strength test showed a significant increase in group D(P<0.01). CT examination showed that BMD of femoral cortical bone increased significantly only in group D(P<0.05).Wnt4 mRNA expression was significantly up-regulated in R group(P<0.05),and significantly up-regulated in D group(P<0.01);Wnt8 and Wnt9 mRNA expression was significantly up-regulated in R group and D group(P<0.01),while GSK3β expression was significantly up-regulated in D group(P<0.05);the expression of β-catenin mRNA was significantly up-regulated in R group and D group(P<0.01),compared with R group,D group also significantly up-regulated(P<0.05);the expression of Runx2 gene was up-regulated significantly in R group(P<0.05),and very significantly in D group(P<0.01). Wnt8 protein expression in R group and D group increased significantly(P<0.01),and that in D group was also significantly higher than that in R group(P<0.01);the expression of β-catenin and Runx2 protein in R group was significantly higher(P<0.05),and that in D group was significantly highe(rP<0.01),and Runx2 protein expression in D group was also significantly higher than that in R group(P<0.05). Conclusions:Jumping exercise can promote endochondral osteogenesis of growth plate and improve bone health through Wnt/β-catenin signaling pathway in growing rats.
Objective:To explore the effects of different exercise modes on the expression of Wnt/β-catenin signaling pathway-related molecules and endochondral osteogenesis in growing rats. Methods:Growing rats were exercised in different ways for 6 weeks. Bone strength was measured by three-point bending,femoral micro-structure was detected by micro-CT,and cytokine gene and protein expression were detected by RT-PCR and western-bloting. Results:Compared with group C,the wet weight of tibia and femur circumference in group D increased significantly(P<0.01),and group D increased significantly(P<0.05). Bone strength test showed a significant increase in group D(P<0.01). CT examination showed that BMD of femoral cortical bone increased significantly only in group D(P<0.05).Wnt4 mRNA expression was significantly up-regulated in R group(P<0.05),and significantly up-regulated in D group(P<0.01);Wnt8 and Wnt9 mRNA expression was significantly up-regulated in R group and D group(P<0.01),while GSK3β expression was significantly up-regulated in D group(P<0.05);the expression of β-catenin mRNA was significantly up-regulated in R group and D group(P<0.01),compared with R group,D group also significantly up-regulated(P<0.05);the expression of Runx2 gene was up-regulated significantly in R group(P<0.05),and very significantly in D group(P<0.01). Wnt8 protein expression in R group and D group increased significantly(P<0.01),and that in D group was also significantly higher than that in R group(P<0.01);the expression of β-catenin and Runx2 protein in R group was significantly higher(P<0.05),and that in D group was significantly highe(rP<0.01),and Runx2 protein expression in D group was also significantly higher than that in R group(P<0.05). Conclusions:Jumping exercise can promote endochondral osteogenesis of growth plate and improve bone health through Wnt/β-catenin signaling pathway in growing rats.
引文
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[21]LIU S,ZHANG E,YANG M,et al.Overexpression of Wnt11 promotes chondrogenic differentiation of bone marrow-derived mesenchymal stem cells in synergism with TGF-β[J].Molecular&Cellular Biochemistry,2014,390(1-2):123-131.
[22]STUDER D,MILLAN C,?ZTüRK E,et al.Molecular and biophysical mechanisms regulating hypertrophic differentiation in chondrocytes and mesenchymal stem cells[J].European Cells&Materials,2012,24(7):118.
[23]BRADLEY E W,DRISSI M H.WNT5A regulates chondrocyte differentiation through differential use of the CaN/NFAT and IKK/NF-kappaBpathways[J].Molecular Endocrinology,2010,24(8):1581.
[24]LING I T,ROCHARD L,LIAO E C.Distinct requirements of wls,wnt9a,wnt5b and gpc4 in regulating chondrocyte maturation and timing of endochondral ossification[J].Developmental Biology,2017,421(2):219-232.
[25]LU C,WAN Y,CAO J,et al.Wnt-mediated reciprocal regulation between cartilage and bone development during endochondral ossification[J].Bone,2013,53(2):566-574.
[26]ZHONG L,HUANG X,KARPERIEN M,et al.The Regulatory Role of Signaling Crosstalk in Hypertrophy of MSCs and Human Articular Chondrocytes[J].International Journal of Molecular Sciences,2015,16(8):19225-19247.
[2]AMORIM T,DUR?ES C,MACHADO J C,et al.Genetic variation in Wnt/β-catenin and ER signalling pathways in female and male elite dancers and its associations with low bone mineral density:a cross-section and longitudinal study[J].Osteoporosis international,2018(6):1-14.
[3]KIEL D P,FERRARI S L,CUPPLES L A,et al.Genetic variation at the low-density lipoprotein receptor-related protein 5(LRP5)locus modulates Wnt signaling and the relationship of physical activity with bone mineral density in men[J].Bone(New York),2007,40(3):0-596.
[4]LIU S,ZHOU P U,ZHANG Y.Abnormal expression of key genes and proteins in the canonical Wnt/β-catenin pathway of articular cartilage in a rat model of exercise-induced osteoarthritis[J].Molecular Medicine Reports,2016,13(3):1999-2006.
[5]XI C,LIHUI L,JIANMIN G,et al.Treadmill running exercise prevents senile osteoporosis and upregulates the Wnt signaling pathway in SAMP6 mice[J].Oncotarget,2016,7(44):71072-71086.
[6]RIKA YASUHARA,YOICHI OHTA,TAKAHITO YUASA,et al.Roles ofβ-catenin signaling in phenotypic expression and proliferation of articular cartilage superficial zone cells[J].Laboratory investigation;a journal of technical methods and pathology,2011,91(12):1739-52.
[7]PASOLD J,OSTERBERG A,PETERS K,et al.Reduced expression of Sfrp1 during chondrogenesis and in articular chondrocytes correlates with osteoarthritis in STR/ort mice[J].Experimental Cell Research,2013,319(5):649-59.
[8]NILSSON M,SUNDH D,OHLSSON C,et al.Exercise During Growth and Young Adulthood Is Independently Associated With Cortical Bone Size and Strength in Old Swedish Men[J].Journal of Bone and Mineral Research,2014,29(8):1795-1804.
[9]LAPAUW B M,TAES Y,BOGAERT V,et al.Serum estradiol is associated with volumetric BMD and modulates the impact of physical activity on bone size at the age of peak bone mass:a study in healthy male siblings[J].Journal of Bone&Mineral Research,2010,24(6):1075-1085.
[10]KARSENTY G,OURY F.Biology without walls:the novel endocrinology of bone[J].Annual Review of Physiology,2012,74(1):87.
[11]赵常红,李世昌,孙朋,等.运动对骨骼和肌肉的共调作用研究[J].首都体育学院学报,2017(6):565-570.
[12]IWAMOTO J,YEH J K,ALOIA J F.Effect of Deconditioning on Cortical and Cancellous Bone Growth in the Exercise Trained Young Rats[J].Journal of Bone&Mineral Research,2010,15(9):1842-1849.
[13]赵常红,李世昌,孙朋,等.运动力学刺激对骨质量影响的研究进展[J].中国骨质疏松杂志,2018,24(06):820-825.
[14]ZHONG L,HUANG X,KARPERIEN M,et al.The Regulatory Role of Signaling Crosstalk in Hypertrophy of MSCs and Human Articular Chondrocytes[J].International Journal of Molecular Sciences,2015,16(8):19225-19247.
[15]DUESTERDIECKZELLMER K,SEMEVOLOS S,KINSLEY M,et al.Age-related differential gene and protein expression in postnatal cartilage canal and osteochondral junction chondrocytes[J].Gene Expression Patterns,2015,17(1):1-10.
[16]STUDER D,MILLAN C,?ZTüRK E,et al.Molecular and biophysical mechanisms regulating hypertrophic differentiation in chondrocytes and mesenchymal stem cells[J].European Cells and Materials,2012;24:118-35.
[17]杨念恩.不同方式运动对生长期小鼠骨合成代谢和Wnt信号通路的影响[D].上海:华东师范大学,2014.
[18]ENOMOTO-IWAMOTO M,KITAGAKI J,KOYAMA E,et al.The Wnt antagonist Frzb-1 regulates chondrocyte maturation and long bone development during limb skeletogenesis[J].Developmental Biology,2002,251(1):142-56.
[19]CHURCH V,NOHNO T,LINKER C,et al.Wnt regulation of chondrocyte differentiation[J].Journal of Cell Science,2002,115(24):4809-18.
[20]HARTMANN C,TABIN C J.Wnt-14 plays a pivotal role in inducing synovial joint formation in the developing appendicular skeleton[J].Cell,2001,104(3):341-51.
[21]LIU S,ZHANG E,YANG M,et al.Overexpression of Wnt11 promotes chondrogenic differentiation of bone marrow-derived mesenchymal stem cells in synergism with TGF-β[J].Molecular&Cellular Biochemistry,2014,390(1-2):123-131.
[22]STUDER D,MILLAN C,?ZTüRK E,et al.Molecular and biophysical mechanisms regulating hypertrophic differentiation in chondrocytes and mesenchymal stem cells[J].European Cells&Materials,2012,24(7):118.
[23]BRADLEY E W,DRISSI M H.WNT5A regulates chondrocyte differentiation through differential use of the CaN/NFAT and IKK/NF-kappaBpathways[J].Molecular Endocrinology,2010,24(8):1581.
[24]LING I T,ROCHARD L,LIAO E C.Distinct requirements of wls,wnt9a,wnt5b and gpc4 in regulating chondrocyte maturation and timing of endochondral ossification[J].Developmental Biology,2017,421(2):219-232.
[25]LU C,WAN Y,CAO J,et al.Wnt-mediated reciprocal regulation between cartilage and bone development during endochondral ossification[J].Bone,2013,53(2):566-574.
[26]ZHONG L,HUANG X,KARPERIEN M,et al.The Regulatory Role of Signaling Crosstalk in Hypertrophy of MSCs and Human Articular Chondrocytes[J].International Journal of Molecular Sciences,2015,16(8):19225-19247.