单侧前牙反对大鼠下颌髁突软骨内CaR、PTHrP表达的影响研究
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摘要
目的探索单侧前牙反(unilateral anterior crossbite,UAC)对大鼠下颌髁突软骨内钙离子敏感受体(calcium-sensing receptor,CaR)、甲状旁腺激素相关蛋白(parathyroid hormone-related protein,PTHrP)分子表达以及软骨细胞增殖和发育的影响。方法选择6周龄雌性SD大鼠48只,根据实验时间点(2、4、8周)随机分为3大组,每大组再随机分为2组(对照组、UAC实验组),每组动物数量8只。对SD大鼠施加UAC刺激,于2、4、8周后分别处死各组大鼠,对颞下颌关节软骨进行CaR、PTHrP、PTHrP受体(PPR)分子的免疫组化染色,提取软骨内mRNA进行增殖标志分子、软骨细胞表型分子和终末分化相关分子的Real-time PCR检测。结果 UAC实验组大鼠髁突软骨2周时即开始出现CaR的表达增高,4周时继发有PTHrP的表达上调,但是其惟一受体PPR的表达从2周时即显著降低;增殖标志分子和软骨细胞表型分子的表达从2周时开始显著降低,终末分化相关分子的表达从4周开始显著增高。结论 UAC可导致下颌髁突软骨内CaR、PTHrP表达改变,进而影响软骨细胞的增殖和分化状态。
Objective To investigate the effects of unilateral anterior crossbite on the expression of CaR,PTHrP and the proliferation and differentiation of chondrocytes in the mandibular condylar cartilage of rats. Methods Forty-eight 6-weeks-old female SD rats were randomly divided into 3 groups according to the experimental time point(2, 4 and 8 weeks). Each group was then randomly divided into 2 subgroups(control and UAC subgroup), with 8 animals in each subgroup. UAC stimulation was applied to rats, which were killed 2, 4 and 8 weeks later. Expression of CaR,PTHrP and PPR in mandibular condylar cartilage were detected by immunohistochemical staining. mRNA of proliferation markers and differentiation markers in cartilage was detected by real-time PCR. Results CaR expression increased at 2 weeks and PTHrP expression increased at 4 weeks. But expression of PPR, the only receptor of PTHrP, decreased significantly at 2 weeks. The expression of proliferation markers and phenotype markers of chondrocyte decreased significantly from 2 weeks, and the expression of terminal differentiation markers increased significantly from 4 weeks. Conclusion Conclusion The unilateral anterior crossbite can induce the changes of CaR and PTHrP expression in the mandibular condylar cartilage, and thereby affect the proliferation and differentiation of chondrocytes.
Objective To investigate the effects of unilateral anterior crossbite on the expression of CaR,PTHrP and the proliferation and differentiation of chondrocytes in the mandibular condylar cartilage of rats. Methods Forty-eight 6-weeks-old female SD rats were randomly divided into 3 groups according to the experimental time point(2, 4 and 8 weeks). Each group was then randomly divided into 2 subgroups(control and UAC subgroup), with 8 animals in each subgroup. UAC stimulation was applied to rats, which were killed 2, 4 and 8 weeks later. Expression of CaR,PTHrP and PPR in mandibular condylar cartilage were detected by immunohistochemical staining. mRNA of proliferation markers and differentiation markers in cartilage was detected by real-time PCR. Results CaR expression increased at 2 weeks and PTHrP expression increased at 4 weeks. But expression of PPR, the only receptor of PTHrP, decreased significantly at 2 weeks. The expression of proliferation markers and phenotype markers of chondrocyte decreased significantly from 2 weeks, and the expression of terminal differentiation markers increased significantly from 4 weeks. Conclusion Conclusion The unilateral anterior crossbite can induce the changes of CaR and PTHrP expression in the mandibular condylar cartilage, and thereby affect the proliferation and differentiation of chondrocytes.
引文
[1]Scrivani SJ,Keith DA,Kaban LB.Temporomandibular disorders[J].N Engl J Med,2008,359(25):2693-2705.
[2]张婧.实验性咬合紊乱致大鼠TMJ髁突骨关节炎样变及Cx43半通道在关节软骨退变中的作用[D].西安:第四军医大学,2013.
[3]Ba J,Friedman PA.Calcium-sensing receptor regulation of renal mineral ion transport[J].Cell Calcium,2004,35(3):229-237.
[4]Aslanova UF,Morimoto T,Farajov EI,et al.Chloride-dependent intracellular pH regulation via extracellular calcium-sensing receptor in the medullary thick ascending limb of the mouse kidney[J].Tohoku J Exp Med,2006,210(4):291-300.
[5]Chattopadhyay N,Legradi G,Bai M,et al.Calcium-sensing receptor in the rat hippocampus:a developmental study[J].Brain Res Dev Brain Res,1997,100(1):13-21.
[6]Bandyopadhyay S,Jeong KH,Hansen JT,et al.Calcium-sensing receptor stimulates secretion of an interferon-gamma-induced monokine(CXCL10)and monocyte chemoattractant protein-3in immortalized GnRH neurons[J].J Neurosci Res,2007,85(4):882-895.
[7]Chang W,Tu C,Chen TH,et al.Expression and signal transduction of calcium-sensing receptors in cartilage and bone[J].Endocrinology,1999,140(12):5883-5893.
[8]Chang W,Tu C,Chen TH,et al.The extracellular calcium-sensing receptor(CaSR)is a critical modulator of skeletal development[J].Sci Signal,2008,1(35):ra1.
[9]Lee K,Lanske B,Karaplis AC,et al.Parathyroid hormone-related peptide delays terminal differentiation of chondrocytes during endochondral bone development[J].Endocrinology,1996,137(11):5109-5118.
[10]Chen J,Gupta T,Barasz JA,et al.Analysis of microarchitectural changes in a mouse temporomandibular joint osteoarthritis model[J].Arch Oral Biol,2009,54(12):1091-1098.362356
[11]Terkeltaub R,Lotz M,Johnson K,et al.Parathyroid hormonerelated proteins is abundant in osteoarthritic cartilage,and the parathyroid hormone-related protein 1-173 isoform is selectively induced by transforming growth factor beta in articular chondrocytes and suppresses generation of extracellular inorganic pyrophosphate[J].Arthritis Rheum,1998,41(12):2152-2164.
[12]Liu YD,Liao LF,Zhang HY,et al.Reducing dietary loading decreases mouse temporomandibular joint degradation induced by anterior crossbite prosthesis[J].Osteoarthritis Cartilage,2014,22(2):302-312.
[13]Wang YL,Zhang J,Zhang M,et al.Cartilage degradation in temporomandibular joint induced by unilateral anterior crossbite prosthesis[J].Oral Dis,2014,20(3):301-306.
[14]Zhang J,Zhang HY,Zhang M,et al.Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage[J].Osteoarthritis Cartilage,2014,22(6):822-830.
[15]Burton DW,Foster M,Johnson KA,et al.Chondrocyte calciumsensing receptor expression is up-regulated in early guinea pig knee osteoarthritis and modulates PTHrP,MMP-13,and TIMP-3 expression[J].Osteoarthritis Cartilage,2005,13(5):395-404.
[16]Robinson DL,Kersh ME,Walsh NC,et al.Mechanical properties of normal and osteoarthritic human articular cartilage[J].JMech Behav Biomed Mater,2016,61:96-109.
[17]Ruggiero L,Zimmerman BK,Park M,et al.Roles of the fibrous superficial zone in the mechanical behavior of tmj condylar cartilage[J].Ann Biomed Eng,2015,43(11):2652-2662.
[18]Hilal G,Massicotte F,Martel-Pelletier J,et al.Endogenous prostaglandin E2 and insulin-like growth factor 1 can modulate the levels of parathyroid hormone receptor in human osteoarthritic osteoblasts[J].J Bone Miner Res,2001,16(4):713-721.
[2]张婧.实验性咬合紊乱致大鼠TMJ髁突骨关节炎样变及Cx43半通道在关节软骨退变中的作用[D].西安:第四军医大学,2013.
[3]Ba J,Friedman PA.Calcium-sensing receptor regulation of renal mineral ion transport[J].Cell Calcium,2004,35(3):229-237.
[4]Aslanova UF,Morimoto T,Farajov EI,et al.Chloride-dependent intracellular pH regulation via extracellular calcium-sensing receptor in the medullary thick ascending limb of the mouse kidney[J].Tohoku J Exp Med,2006,210(4):291-300.
[5]Chattopadhyay N,Legradi G,Bai M,et al.Calcium-sensing receptor in the rat hippocampus:a developmental study[J].Brain Res Dev Brain Res,1997,100(1):13-21.
[6]Bandyopadhyay S,Jeong KH,Hansen JT,et al.Calcium-sensing receptor stimulates secretion of an interferon-gamma-induced monokine(CXCL10)and monocyte chemoattractant protein-3in immortalized GnRH neurons[J].J Neurosci Res,2007,85(4):882-895.
[7]Chang W,Tu C,Chen TH,et al.Expression and signal transduction of calcium-sensing receptors in cartilage and bone[J].Endocrinology,1999,140(12):5883-5893.
[8]Chang W,Tu C,Chen TH,et al.The extracellular calcium-sensing receptor(CaSR)is a critical modulator of skeletal development[J].Sci Signal,2008,1(35):ra1.
[9]Lee K,Lanske B,Karaplis AC,et al.Parathyroid hormone-related peptide delays terminal differentiation of chondrocytes during endochondral bone development[J].Endocrinology,1996,137(11):5109-5118.
[10]Chen J,Gupta T,Barasz JA,et al.Analysis of microarchitectural changes in a mouse temporomandibular joint osteoarthritis model[J].Arch Oral Biol,2009,54(12):1091-1098.362356
[11]Terkeltaub R,Lotz M,Johnson K,et al.Parathyroid hormonerelated proteins is abundant in osteoarthritic cartilage,and the parathyroid hormone-related protein 1-173 isoform is selectively induced by transforming growth factor beta in articular chondrocytes and suppresses generation of extracellular inorganic pyrophosphate[J].Arthritis Rheum,1998,41(12):2152-2164.
[12]Liu YD,Liao LF,Zhang HY,et al.Reducing dietary loading decreases mouse temporomandibular joint degradation induced by anterior crossbite prosthesis[J].Osteoarthritis Cartilage,2014,22(2):302-312.
[13]Wang YL,Zhang J,Zhang M,et al.Cartilage degradation in temporomandibular joint induced by unilateral anterior crossbite prosthesis[J].Oral Dis,2014,20(3):301-306.
[14]Zhang J,Zhang HY,Zhang M,et al.Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage[J].Osteoarthritis Cartilage,2014,22(6):822-830.
[15]Burton DW,Foster M,Johnson KA,et al.Chondrocyte calciumsensing receptor expression is up-regulated in early guinea pig knee osteoarthritis and modulates PTHrP,MMP-13,and TIMP-3 expression[J].Osteoarthritis Cartilage,2005,13(5):395-404.
[16]Robinson DL,Kersh ME,Walsh NC,et al.Mechanical properties of normal and osteoarthritic human articular cartilage[J].JMech Behav Biomed Mater,2016,61:96-109.
[17]Ruggiero L,Zimmerman BK,Park M,et al.Roles of the fibrous superficial zone in the mechanical behavior of tmj condylar cartilage[J].Ann Biomed Eng,2015,43(11):2652-2662.
[18]Hilal G,Massicotte F,Martel-Pelletier J,et al.Endogenous prostaglandin E2 and insulin-like growth factor 1 can modulate the levels of parathyroid hormone receptor in human osteoarthritic osteoblasts[J].J Bone Miner Res,2001,16(4):713-721.