大鼠正畸牙牙根吸收中骨保护素及其配体mRNA的表达的研究
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摘要
目的:探讨OPG/RANKL/RANK系统在正畸牙牙根吸收中的作用。方法:选用12周龄雄性SD大鼠30只。采用自身对照,右侧为实验组,左侧为对照组。在右上颌第一磨牙加力100g,分别在加力0天、1天、4天、8天、12天后处死大鼠各6只。通过HE染色及扫描电镜观察牙根吸收情况,TRAP染色法破骨细胞数量,原位杂交法检测OPGmRNA和RANKLmRNA的表达.结果:1.HE染色发现8天后第一磨牙近中根出现明显的牙根吸收,到达牙本质浅层。2.扫描电镜发现在第4天开始出现牙根吸收,在第8天及第12天时出现大面积的牙根吸收。3.牙槽骨吸收陷窝中的破骨细胞在4天时达到高峰,在8天和12天时数量下降。牙骨质吸收陷窝中破牙骨质细胞在4天时开始出现,8天和12天时持续增高。4. OPGmRNA表达量在张力侧显著增加,8天时达到高峰,主要表达于成纤维细胞中,在成牙骨质细胞和成牙本质细胞中也有表达。5. RANKLmRNA表达量在压力侧显著增加,4天时达到高峰,主要表达于成纤维细胞、破骨细胞、破牙骨质细胞中,在成牙骨质细胞中也有表达。6. RANKLmRNA/OPGmRNA在压力侧显著增加。结论:压力侧RANKLmRNA的表达量以及RANKLmRNA/OPGmRNA随牙根吸收程度及破(牙)骨细胞的数量的增加而增加,说明OPG/RANKL/RANK系统在牙根吸收中起着重要的作用。
Objective: To approach the role of OPG/RANKL/RANK system in the root resorption of orthodontic tooth. Methods: Thirty twelve-month-old male SD rats were studied. Use the method of own control, right side was the experimental group, left side was the control group. 100g force was loaded on the right first molar, rats were killed after 0 day,1 day,2 days,4 days,8 days and 12 days’force loaded. The root resorption of mesial root of first molar, the quantity of osteoclasts and the expression of OPGmRNA and RANKLmRNA were observed by histological section and scanning electron micrograph, TRAP staining and in-situ hybridization. Results: 1.A serious resorption appeared at the pressure side of the mesial root at day 8, which reached shallow of the dentin. 2. From the scanning electron micrograph, at day 4, small area of root resorption appeared. At days 8 and 12, large area of resorption can be seen. 3. The peak quantity of the osteoclasts, which located in the alveolar resorption lacunaes, appeared at day 4, and at days 8 and 12, the quatity of these osteoclasts went down. The osteoclasts in the dentin resorption lacunae appeared at day 4, and at days 8 and 12 increased continuously. 4. The expression of OPGmRNA was significantly increased in the tension side. At day 4, it reached the peak. OPGmRNA was expressed in fibroblasts,odontoblasts and cementoblasts. 5. The expression of RANKLmRNA was significantly increased in the pressure side. At day 4, it reached the peak. RANKLmRNA was expressed in osteoclasts, cementoclasts, fibroblasts and cementoblasts. 6. The ratio of RANKLmRNA and OPGmRNA was significantly increased in pressure side. Conclusions: The increase of RANKLmRNA and the ratio of the RANKLmRNA and OPGmRNA followed the increase of the extent of root resorption and the quantity of osteoclasts and cementoclasts, so OPG/RANKL system played an important role in root resorption.
引文
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    [1]. Schoppet M,Preiddner KT,Hofbauer LC.RANK lifand and osteoprotegerin: paracrine regulators of bone metabolism and vascular function[J].Arterioscler Thromb Vasc Biol,2002;22:549-553.
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    [7]. Yasuda H,Shima N,Nakagawa N et al.Identity of osteoclastogenesisi inhibitory factor and osteoprotegerin:a mechanism by which OPG/OCIF inhibited osteoclastogenensis in vitro[J].Endocrinology,1998;39:1329-1337.
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    [21]. Udagawa N,Takahashi N,Jimi E,Matsuzaki ea tl.Osteoblasts/stromal cells stimulate osteoclast activation through expression of osteoclast differentiation factor RANKL but not macrophage colony-stimulating factor[J].Bone,1999;25:517-523.
    [22]. Shiotani A,Shibasaki Y,Sasaki T.Localization of receptor activator of NF kappa B ligand,RANKL,in periodontal tissues during experiment movement of rat molar[J].J Electron Microsc(Tokyo),2001;50:365-369.
    [23]. Ogasawara T,Yoshimine Y,Kiyoshima T ea tl.In situ expression of RANKL,RANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue[J]. J Periodont Res, 2004;39:42-49.
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    [27]. Oshro T,Shibasaki Y,Martin J ea tl.Immunolocalization of vacuolar-type H+-ATPase,cathesin K,matrix metalloproteinase-9,and receptor activator of NFΚβligand in odontoclasts during physiological root resorption of human deciduous teeth[J].Anat Rec,2001;264:305-311.
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