负重爬梯运动调节去卵巢大鼠OPG/Ikkα/Runx2基因表达以及对股骨结构和生物力学的影响
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摘要
目的:探讨负重爬梯运动对去卵巢大鼠OPG/Ikkα/Runx2基因表达调控以及对股骨微结构与生物力学特征变化的影响。方法:将30只SD雌性大鼠随机分为3组,假手术组(CON组,10只)、去卵巢手术组(OVX组,10只)和去卵巢手术运动组(OVX-CL组,10只)。OVX-CL组进行85°、梯阶间距2 cm、总高1 m、负重量为自身体重15%的爬梯训练。8周实验干预后称重,并检测血清中磷(P)、钙(Ca)、碱性磷酸酶(ALP)、抗酒石酸性磷酸酶(StraCP)、破骨细胞分化因子(RANKL)、骨保护素(OPG)等含量;qRT-PCR检测RANKL、OPG、IkB激酶α(Ikkα)、核因子κB2(NFκB2)和Runt相关转录因子2(Runx2)表达量,并对股骨的微结构参数(Tb.N、Tb.Th、Tb.Sp、BMD~(trab)、BMD~(cort)、BV/TV、和SMI)和生物力学特性(ML、Stiffness、EAC和MOE)进行检测。结果:OVX-CL组血清中钙(Ca)和RANKL含量均低于OVX组(P<0.05),ALP水平与OPG/RANKL比值均高于OVX组(P<0.05);OVX-CL组股骨组织RANKL、Ikkα及NFκB2的基因表达量与OVX组相比降低(P<0.05),OVX-CL组与OVX组相比,OPG和Runx2基因的表达量增加(P<0.01);MicroCT检测结果显示,OVX-CL组与OVX组相比,骨小梁数量与松质骨矿密度增加(P<0.05),骨小梁分离度、结构模型指数均降低(P<0.05);OVX-CL组最大载荷及刚度均增加(P<0.05)。结论:负重爬梯运动可通过对OPG/Ikkα/Runx2的基因调控改善成骨与破骨之间的平衡状态,对提高去卵巢大鼠骨微结构和力学特征有积极作用。
Objective To investigate the effect of weight-bearing climbing-ladder exercise on the expression of the osteoprotegerin(OPG)/IkB kinase α(Ikkα)/Runtgene,the changes of femur microstructure and biomechanical properties of ovariectomized rats.Methods Thirty Sprague-Dawley female rats were randomly divided into a sham operation(CON) group,an ovariectomy(OVX) group and an ovariectomy exercise(OVXCL group underwent a training of climbing a 1-meter ladder with an 85°-angle slope and 2-cm step spacing,with 15% of their own body-weight mass added. After an 8-week training, the serum levels of phosphorus(P),calcium(Ca),alkaline phosphatase(ALP),tartrate-resistant acid phosphatase(StraCP),osteoclast differentiation factor(RANKL) and osteoprotegerin(OPG) were measured. The gene expression of RANKL,OPG,Ikkα,nuclear factor kappa B2(NF kappa B2) and Runx2 was detected using the quantitative reverse transcription polymerase chain reaction and the parameters of microstructure of the femur(Tb. N, Tb. Th, Tb. Sp, BMD~(trab), BMD~(cort), BV/TV, and SMI) and biomechanical properties(ML, Stiffness, EAC and MOE) were measured.Results The average serum calcium and RANKL of the OVX-CL group were significantly lower than the OVX group(P<0.05),while the ALP level and the ratio of OPG/RANKL were significantly higher than the OVX group(P<0.05). The gene expression levels of RANKL, Ikkα and NFκB2 decreased significantly in the OVXCL group compared with the OVX group(P<0.05),while that of the OPG and Runx2 increased significantly in the OVX-CL group compared with the OVX group(P<0.01). The result of Micro-CT showed that,compared with the OVX group,the trabeculae number and mineral density of the cancellous bone of the femoral head increased significantly in the OVX-CL group(P<0.05), while the trabeculae separation and structural model index decreased significantly(P<0.05). The maximum loading and stiffness of the OVX-CL group also increased(P<0.05).Conclusions The weight-bearing climbing ladder exercise,which can improve the balance between the osteogenesis and osteoclast through the gene regulation of OPG/Ikkα/Runx2,has a positive effect on improving the bone microstructure and mechanical properties of ovariectomized rats.
Objective To investigate the effect of weight-bearing climbing-ladder exercise on the expression of the osteoprotegerin(OPG)/IkB kinase α(Ikkα)/Runtgene,the changes of femur microstructure and biomechanical properties of ovariectomized rats.Methods Thirty Sprague-Dawley female rats were randomly divided into a sham operation(CON) group,an ovariectomy(OVX) group and an ovariectomy exercise(OVXCL group underwent a training of climbing a 1-meter ladder with an 85°-angle slope and 2-cm step spacing,with 15% of their own body-weight mass added. After an 8-week training, the serum levels of phosphorus(P),calcium(Ca),alkaline phosphatase(ALP),tartrate-resistant acid phosphatase(StraCP),osteoclast differentiation factor(RANKL) and osteoprotegerin(OPG) were measured. The gene expression of RANKL,OPG,Ikkα,nuclear factor kappa B2(NF kappa B2) and Runx2 was detected using the quantitative reverse transcription polymerase chain reaction and the parameters of microstructure of the femur(Tb. N, Tb. Th, Tb. Sp, BMD~(trab), BMD~(cort), BV/TV, and SMI) and biomechanical properties(ML, Stiffness, EAC and MOE) were measured.Results The average serum calcium and RANKL of the OVX-CL group were significantly lower than the OVX group(P<0.05),while the ALP level and the ratio of OPG/RANKL were significantly higher than the OVX group(P<0.05). The gene expression levels of RANKL, Ikkα and NFκB2 decreased significantly in the OVXCL group compared with the OVX group(P<0.05),while that of the OPG and Runx2 increased significantly in the OVX-CL group compared with the OVX group(P<0.01). The result of Micro-CT showed that,compared with the OVX group,the trabeculae number and mineral density of the cancellous bone of the femoral head increased significantly in the OVX-CL group(P<0.05), while the trabeculae separation and structural model index decreased significantly(P<0.05). The maximum loading and stiffness of the OVX-CL group also increased(P<0.05).Conclusions The weight-bearing climbing ladder exercise,which can improve the balance between the osteogenesis and osteoclast through the gene regulation of OPG/Ikkα/Runx2,has a positive effect on improving the bone microstructure and mechanical properties of ovariectomized rats.
引文
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[40]Huang W,Yang S,Shao J,et al.Signaling and transcriptional regulation in osteoblast commitment and differentiation[J].Front Biosci,2007,12:3068-3092.
[41]Hu N,Feng C,Jiang Y,et al.Regulative effect of Mir-205 on osteogenic differentiation of bone mesenchymal stem cells(BMSCs):Possible role of SATB2/Runx2and ERK/MAPK pathway[J].Int J Mol Sci,2015,16(5):10491-10506.
[42]Chang J,Wang Z,Tang E,et al.Inhibition of osteoblastic bone formation by nuclear factor-kappaB[J].Nat Med,2009,15(6):682-689.
[43]Notomi T,Karasaki I,Okazaki Y,et al.Insulinogenic sucrose+amino acid mixture ingestion immediately after resistance exercise has an anabolic effect on bone compared with non-insulinogenic fructose+amino acid mixture in growing rats[J].Bone,2014,65:42-48.
[44]Stringhetta-Garcia CT,Singulani MP,Santos LF,et al.The effects of strength training and raloxifene on bone health in aging ovariectomized rats[J].Bone,2016,85:45-54.
[45]Pacios S,Xiao W,Mattos M,et al.Osteoblast lineage cells play an essential role in periodontal bone loss through activation of nuclear factor-kappa B[J].Sci Rep,2015,5:16694.
[46]Khedgikar V,Kushwaha P,Gautam J,et al.Withaferin A:a proteasomal inhibitor promotes healing after injury and exerts anabolic effect on osteoporotic bone[J].Cell Death Dis,2013,4:e778.
[47]Ma X,Meng J,Jia M,et al.Exendin-4,a glucagonlike peptide-1 receptor agonist,prevents osteopenia by promoting bone formation and suppressing bone resorption in aged ovariectomized rats[J].J Bone Miner Res,2013,28(7):1641-1652.
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[2]Hsieh YF,Turner CH.Effects of loading frequency on mechanically induced bone formation[J].J Bone Miner Res,2001,16(5):918-924.
[3]Levine JP.Pharmacologic and nonpharmacologic management of osteoporosis[J].Clin Cornerstone,2006,8(1):40-53.
[4]Kelley GA.Exercise and regional bone mineral density in postmenopausal women:a meta-analytic review of randomized trials[J].Am J Phys Med Rehabil,1998,77(1):76-87.
[5]Seeman E.An exercise in geometry[J].J Bone Miner Res,2002,17(3):373-380.
[6]Menkes A,Mazel S,Redmond RA,et al.Strength training increases regional bone mineral density and bone remodeling in middle-aged and older men[J].J Appl Physiol(1985),1993,74(5):2478-2484.
[7]Fujimura R,Ashizawa N,Watanabe M,et al.Effect of resistance exercise training on bone formation and resorption in young male subjects assessed by biomarkers of bone metabolism[J].J Bone Miner Res,1997,12(4):656-662.
[8]Danz AM,Zittermann A,Schiedermaier U,et al.The effect of a specific strength-development exercise on bone mineral density in perimenopausal and postmenopausal women[J].J Womens Health(Larchmt),1998,7(6):701-709.
[9]Bradney M,Pearce G,Naughton G,et al.Moderate exercise during growth in prepubertal boys:changes in bone mass,size,volumetric density,and bone strength:a controlled prospective study[J].J Bone Miner Res,1998,13(12):1814-1821.
[10]Nicholson VP,McKean MR,Burkett BJ.Low-load highrepetition resistance training improves strength and gait speed in middle-aged and older adults[J].J Sci Med Sport,2015,18(5):596-600.
[11]Van Roie E,Delecluse C,Coudyzer W,et al.Strength training at high versus low external resistance in older adults:effects on muscle volume,muscle strength,and force-velocity characteristics[J].Exp Gerontol,2013,48(11):1351-1361.
[12]Vincent KR,Braith RW.Resistance exercise and bone turnover in elderly men and women[J].Med Sci Sports Exerc,2002,34(1):17-23.
[13]Kerr D,Morton A,Dick I,et al.Exercise effects on bone mass in postmenopausal women are site-specific and load-dependent[J].J Bone Miner Res,1996,11(2):218-225.
[14]Bemben DA,Bemben MG.Dose-response effect of 40weeks of resistance training on bone mineral density in older adults[J].Osteoporos Int,2011,22(1):179-186.
[15]Basat H,Esmaeilzadeh S,Eskiyurt N.The effects of strengthening and high-impact exercises on bone metabolism and quality of life in postmenopausal women:a randomized controlled trial[J].J Back Musculoskelet Rehabil,2013,26(4):427-435.
[16]Johnson ML,Kamel MA.The Wnt signaling pathway and bone metabolism[J].Curr Opin Rheumatol,2007,19(4):376-382.
[17]Amarasekara DS,Yu J,Rho J.Bone Loss Triggered by the Cytokine Network in Inflammatory Autoimmune Diseases[J].J Immunol Res,2015,2015:832127.
[18]Seibel MJ.Biochemical markers of bone turnover:part I:biochemistry and variability[J].Clin Biochem Rev,2005,26(4):97-122.
[19]Meier C,Seibel MJ,Kraenzlin ME.Use of bone turnover markers in the real world:are we there yet?[J].JBone Miner Res,2009,24(3):386-388.
[20]Garnero P,Delmas PD.Assessment of the serum levels of bone alkaline phosphatase with a new immunoradiometric assay in patients with metabolic bone disease[J].JClin Endocrinol Metab,1993,77(4):1046-1053.
[21]卜淑敏,杨涵,韩天雨.不同干预疗法对去卵巢骨质疏松大鼠骨微结构影响的对比研究[J].中国运动医学杂志,2016,35(03):257-261.
[22]Tang L,Gao X,Yang X,et al.Ladder-climbing training prevents bone loss and microarchitecture deterioration in diet-induced obese rats[J].Calcif Tissue Int,2016,98(1):85-93.
[23]张林.不同强度运动对骨质疏松大鼠骨生物力学性能的影响[J].体育科学,2000,(5):72-76.
[24]Oh T,Tanaka S,Naka T,et al.Effects of high-intensity swimming training on the bones of ovariectomized rats[J].J Exerc Nutrition Biochem,2016,20(3):39-45.
[25]Hernandes L,Ramos AL,Micheletti KR,et al.Densitometry,radiography,and histological assessment of collagen as methods to evaluate femoral bones in an experimental model of osteoporosis[J].Osteoporos Int,2012,23(2):467-473.
[26]Pighon A,Gutkowska J,Jankowski M,et al.Exercise training in ovariectomized rats stimulates estrogenic-like effects on expression of genes involved in lipid accumulation and subclinical inflammation in liver[J].Metabolism,2011,60(5):629-639.
[27]Smith MZ,Goettsch BM,Van Ramshorst RD,et al.Resistance training and bone mineral density during growth[J].Int J Sports Med,2008,29(4):316-321.
[28]Pierce RA,Lee LC,Ahles CP,et al.Different training volumes yield equivalent increases in BMD[J].Int JSports Med,2010,31(11):803-809.
[29]Kayser BD,Godfrey JK,Cunningham RM,et al.Equal BMD after daily or triweekly exercise in growing rats[J].Int J Sports Med,2010,31(1):44-50.
[30]Joo W,Singh H,Ahles CP,et al.Training-induced increase in bone mineral density between growing male and female rats[J].Int J Sports Med,2015,36(12):992-998.
[31]Dror AD,Virk K,Lee K,et al.Resistance training threshold for elevating bone mineral density in growing female rats[J].Int J Sports Med,2018,39(5):382-389.
[32]Fleck SJ,Kraemer W.Designing resistance training programs,4E[M].Human kinetics,2014.
[33]曹鹏.不同强度跑台运动对老龄雌性大鼠骨量及骨代谢的影响[J].北京体育大学学报,2009,(2):74-76.
[34]唐量,亢依婷,尹博,等.负重爬梯与有氧跑台运动对糖尿病大鼠学习记忆能力的影响及其机制探讨[J].中国应用生理学杂志,2017,33(5):436-440.
[35]Livak KJ,Schmittgen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method[J].Methods,2001,25(4):402-408.
[36]唐量,高小航,赵新娟,等.负重爬梯与负重跑台运动对大鼠股骨微结构及力学特性的影响[J].陕西师范大学学报(自然科学版),2015,(6):100-105.
[37]Milat F,Ng KW.Is Wnt signalling the final common pathway leading to bone formation?[J].Mol Cell Endocrinol,2009,310(1-2):52-62.
[38]Bord S,Ireland DC,Beavan SR,et al.The effects of estrogen on osteoprotegerin,RANKL,and estrogen receptor expression in human osteoblasts[J].Bone,2003,32(2):136-141.
[39]Pichler K,Loreto C,Leonardi R,et al.RANKL is downregulated in bone cells by physical activity(treadmill and vibration stimulation training)in rat with glucocorticoid-induced osteoporosis[J].Histol Histopathol,2013,28(9):1185-1196.
[40]Huang W,Yang S,Shao J,et al.Signaling and transcriptional regulation in osteoblast commitment and differentiation[J].Front Biosci,2007,12:3068-3092.
[41]Hu N,Feng C,Jiang Y,et al.Regulative effect of Mir-205 on osteogenic differentiation of bone mesenchymal stem cells(BMSCs):Possible role of SATB2/Runx2and ERK/MAPK pathway[J].Int J Mol Sci,2015,16(5):10491-10506.
[42]Chang J,Wang Z,Tang E,et al.Inhibition of osteoblastic bone formation by nuclear factor-kappaB[J].Nat Med,2009,15(6):682-689.
[43]Notomi T,Karasaki I,Okazaki Y,et al.Insulinogenic sucrose+amino acid mixture ingestion immediately after resistance exercise has an anabolic effect on bone compared with non-insulinogenic fructose+amino acid mixture in growing rats[J].Bone,2014,65:42-48.
[44]Stringhetta-Garcia CT,Singulani MP,Santos LF,et al.The effects of strength training and raloxifene on bone health in aging ovariectomized rats[J].Bone,2016,85:45-54.
[45]Pacios S,Xiao W,Mattos M,et al.Osteoblast lineage cells play an essential role in periodontal bone loss through activation of nuclear factor-kappa B[J].Sci Rep,2015,5:16694.
[46]Khedgikar V,Kushwaha P,Gautam J,et al.Withaferin A:a proteasomal inhibitor promotes healing after injury and exerts anabolic effect on osteoporotic bone[J].Cell Death Dis,2013,4:e778.
[47]Ma X,Meng J,Jia M,et al.Exendin-4,a glucagonlike peptide-1 receptor agonist,prevents osteopenia by promoting bone formation and suppressing bone resorption in aged ovariectomized rats[J].J Bone Miner Res,2013,28(7):1641-1652.
[48]Boudenot A,Presle N,Uzbekov R,et al.Effect of interval-training exercise on subchondral bone in a chemically-induced osteoarthritis model[J].Osteoarthritis Cartilage,2014,22(8):1176-1185.
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