S1P/S1PR1通路依赖的压力对破骨前体细胞趋化效应
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摘要
目的:探究S1P/S1PR1通路轴在压力下对破骨前体细胞趋化影响的调节作用。方法:建立体外压力模型,通过实时荧光定量聚合酶链式反应,免疫印迹实验和细胞免疫荧光检测压力下Raw264.7细胞SPHK1和S1PR1的mRNA水平和蛋白水平。采用Transwell 24孔板通过趋化实验探究S1P/S1PR1在压力下对Raw264.7细胞的迁移能力的影响。结果:发现0.5、1.0和2.0g/cm2压力作用下,Raw264.7细胞的SPHK1和S1PR1的表达及Raw264.7细胞的趋化显著降低。在S1PR1受体功能性激动剂FTY720作用下,压力作用下趋化能力被抑制的Raw264.7细胞趋化水平显著增加。结论:S1P/S1PR1信号轴在正畸治疗中调节破骨前体细胞迁移与功能发挥重要作用。
Objective:To investigate the role of Sphingosine-1-phosphate/Sphingosine-1-phosphate receptor1(S1P/S1PR1)axis in regulating migration of osteoclast precursors under compressive stress.Methods:Compressed model was built in vitro.The mRNA and protein expression levels of SPHK1 and S1PR1in Raw 264.7cell lines under compressive stress were investigated with real-time PCR,Western-blotting and immunofluoresecence.The regulatory role of S1P/S1PR1 signaling on chemotaxis of Raw 264.7cell lines under compressive stress was explored.Results:Under compressive force of 0.5,1.0and 2.0g/cm2,SPHK1 and S1PR1expressions and chemotaxis of Raw 264.7cell lines were downregulated.And S1P/S1PR1 signaling functional antagonist FTY720 increased the chemotaxis of Raw 264.7cells.Conclusion:S1P/S1PR1 axis regulates migration of osteoclast precursors under compressive stress.
Objective:To investigate the role of Sphingosine-1-phosphate/Sphingosine-1-phosphate receptor1(S1P/S1PR1)axis in regulating migration of osteoclast precursors under compressive stress.Methods:Compressed model was built in vitro.The mRNA and protein expression levels of SPHK1 and S1PR1in Raw 264.7cell lines under compressive stress were investigated with real-time PCR,Western-blotting and immunofluoresecence.The regulatory role of S1P/S1PR1 signaling on chemotaxis of Raw 264.7cell lines under compressive stress was explored.Results:Under compressive force of 0.5,1.0and 2.0g/cm2,SPHK1 and S1PR1expressions and chemotaxis of Raw 264.7cell lines were downregulated.And S1P/S1PR1 signaling functional antagonist FTY720 increased the chemotaxis of Raw 264.7cells.Conclusion:S1P/S1PR1 axis regulates migration of osteoclast precursors under compressive stress.
引文
[1]Huang H,Williams RC,Kyrkanides S.Accelerated orthodontic tooth movement:molecular mechanisms[J].Am J Orthod Dentofacial Orthop,2014,146(5)∶620-632
[2]Kanzaki H,Shinohara F,Itohiya-Kasuya K,et al.Nrf2activation attenuates both orthodontic tooth movement and relapse[J].J Dent Res,2015,94(6)∶787-794
[3]Tsay TP,Chen MH,Oyen OJ.Osteoclast activation and recruitment after application of orthodontic force[J].Am J Orthod Dentofacial Orthop,1999,115(3)∶323-330
[4]杨健,谭颖徽,裘松波.核因子-κB受体活化因子配体在大鼠正畸牙压力侧骨改建中的作用[J].口腔医学研究,2013,20(3)∶259-262
[5]Faccio R,Teitelbaum SL,Fujikawa K,et al.Vav3regulates osteoclast function and bone mass[J].Nat Med,2005,11(3)∶284-290
[6]Prager B,Spampinato SF,Ransohoff RM.Sphingosine 1-phosphate signaling at the blood-brain barrier[J].Trends in molecular medicine,2015,21(6)∶354-363
[7]Emanuel BR,Toshihiro I,Aki K,et al.Effect of PGE2induced by compressive and tensile stresses on cementoblast differentiation in vitro[J].Archives of oral biology,2011,56∶238-246
[8]Ishii T,Shimazu Y,Nishiyama I,et al.The role of sphingosine 1-phosphate in migration of osteoclast precursors;an application of intravital two-photon microscopy[J].Mol Cells,2011,31(5)∶399-403
[9]Ryu J,Kim HJ,Chang EJ,et al.Sphingosine 1-phosphate as a regulator of osteoclast differentiation and osteoclast-osteoblast coupling[J].EMBO J,2006,25(24)∶5840-51
[10]Quint P,Ruan M,Pederson L,et al.Sphingosine 1-phosphate(S1P)receptors 1and 2coordinately induce mesenchymal cell migration through S1Pactivation of complementary kinase pathways[J].J Biol Chem,2013,288(8)∶5398-406
[11]Ishii M,Kikuta J,Shimazu Y,et al.Chemorepulsion by blood S1P regulates osteoclast precursor mobilization and bone remodeling in vivo[J].J Exp Med,2010,207(13)∶2793-2798
[12]Ishii M,Egen JG,Klauschen F,et al.Sphingosine-1-phosphate mobilizes osteoclast precursors and regulates bone homeostasis[J].Nature,2009,458(7237)∶524-528
[2]Kanzaki H,Shinohara F,Itohiya-Kasuya K,et al.Nrf2activation attenuates both orthodontic tooth movement and relapse[J].J Dent Res,2015,94(6)∶787-794
[3]Tsay TP,Chen MH,Oyen OJ.Osteoclast activation and recruitment after application of orthodontic force[J].Am J Orthod Dentofacial Orthop,1999,115(3)∶323-330
[4]杨健,谭颖徽,裘松波.核因子-κB受体活化因子配体在大鼠正畸牙压力侧骨改建中的作用[J].口腔医学研究,2013,20(3)∶259-262
[5]Faccio R,Teitelbaum SL,Fujikawa K,et al.Vav3regulates osteoclast function and bone mass[J].Nat Med,2005,11(3)∶284-290
[6]Prager B,Spampinato SF,Ransohoff RM.Sphingosine 1-phosphate signaling at the blood-brain barrier[J].Trends in molecular medicine,2015,21(6)∶354-363
[7]Emanuel BR,Toshihiro I,Aki K,et al.Effect of PGE2induced by compressive and tensile stresses on cementoblast differentiation in vitro[J].Archives of oral biology,2011,56∶238-246
[8]Ishii T,Shimazu Y,Nishiyama I,et al.The role of sphingosine 1-phosphate in migration of osteoclast precursors;an application of intravital two-photon microscopy[J].Mol Cells,2011,31(5)∶399-403
[9]Ryu J,Kim HJ,Chang EJ,et al.Sphingosine 1-phosphate as a regulator of osteoclast differentiation and osteoclast-osteoblast coupling[J].EMBO J,2006,25(24)∶5840-51
[10]Quint P,Ruan M,Pederson L,et al.Sphingosine 1-phosphate(S1P)receptors 1and 2coordinately induce mesenchymal cell migration through S1Pactivation of complementary kinase pathways[J].J Biol Chem,2013,288(8)∶5398-406
[11]Ishii M,Kikuta J,Shimazu Y,et al.Chemorepulsion by blood S1P regulates osteoclast precursor mobilization and bone remodeling in vivo[J].J Exp Med,2010,207(13)∶2793-2798
[12]Ishii M,Egen JG,Klauschen F,et al.Sphingosine-1-phosphate mobilizes osteoclast precursors and regulates bone homeostasis[J].Nature,2009,458(7237)∶524-528