Tanshinone type IIA inhibits osteoprotegerin and osteoclast differentiation factor expression at relapse stage after orthodontic tooth movement

doi:10.3969/j.issn.2095-4344.2014.11.015

Abstract

Abstract:

BACKGROUND: In recent years, many drugs emerge to control tooth movement, and scholars in China begin to investigate Chinese herbs with moderate nature and small adverse reaction.

OBJECTIVE: To observe the relapse after orthodontic tooth movement, osteoprotegerin and osteoclast differentiation factor expression in periodontal tissue after rats were treated with local tanshinone type IIA at different doses.

METHODS: A total of 48 male Wistar rats were randomly divided into four groups: control, low dose (tanshinone type IIA 0.36 mg/d), medium dose (tanshinone type IIA 0.72 mg/d), and high dose (tanshinone type IIA 1.44 mg/d) groups. Taking anterior teeth as the anchorage, the maxillary first molar of rats was tracted to mesial movement. In experimental groups, gingival mucosa of the first molar was local injected with tanshinone type IIA 1 day

before the force device was removed, while control group was injected with physiological saline, once a day, for

4 weeks. Immediately, 1 week, and 4 weeks after the force device was removed, the distance between the maxillary first molar and second molar was measured and body mass was weighted. The animals were killed after 4 weeks, osteoprotegerin and osteoclast differentiation factor expression in maxillary first molar and periodontal tissue were determined using immunohistochemical staining.

RESULTS AND CONCLUSION: There was no obvious change in the body weight of rats in each group (P > 0.05). In low, medium and high dose groups, recurrent distance of the teeth was shorter than that in control group (P < 0.05), and recurrence percentage was significantly lower than control group (P < 0.05). The greater the dose was, the smaller the degree of recurrence was. Osteoprotegerin expression in the periodontal tissue was significantly higher in the experimental groups than the control group (P < 0.05), while osteoclast differentiation factor expression was significantly lower than the control group (P < 0.05). The ratio of osteoprotegerin/osteoclast differentiation factor in the periodontal tissue was greater than 1 in both control group and experimental groups, and reached the peak in the high dose group. Local delivery of tanshinone type IIA has no impact on body weight of normal rats, and can effectively control the recurrence rate after orthodontic tooth movement. Within a certain range, high dose achieves the most obvious effect. Regulating osteoclast through adjusting the ratio of osteoprotegerin/osteoclast differentiation factor could be the molecular mechanism of tanshinone type IIA accelerating the periodontal tissue rebuilding.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: Salvia miltiorrhiza, orthodontics, osteoprotegerin

CLC Number:

R394.2

Zhang Shi-ying, Liu Ji-guang, Zhao Gang. Tanshinone type IIA inhibits osteoprotegerin and osteoclast differentiation factor expression at relapse stage after orthodontic tooth movement[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(11): 1730-1736.

Figures/Tables 3

2.1 实验动物数量分析实验选用大鼠48只，分为4组，每组12只大鼠，都进入实验数据分析。 2.2 实验前后大鼠体质量的变化由表1可见，实验开始时及给药后1，4周，实验组和对照组大鼠体质量比较差异无显著性意义(P > 0.05)，提示该药物对体质量无明显影响。 2.3 给药后复发百分率对比由表2可见，加力21 d后上颌第1磨牙平均移动了(0.372±0.025) mm，各组间差异无显著性意义(P > 0.05)。给药1，4周后，实验组较对照组牙齿复发距离小，对照组与实验组比较，差异有显著性意义(P < 0.05)；高剂量组抑制复发的程度最明显(P < 0.05)。实验组间比较，高剂量组复发距离最小，低剂量组复发最大；给药4周后，实验组各组间复发百分率差异均有显著性意义(P < 0.05)。 2.4 各组大鼠牙槽骨免疫组织化学染色结果实验动物去除加力装置后，向近中移动的第1磨牙在牙周纤维的牵引下向远中复发移动。选取第1磨牙近中腭根根中部牙槽骨及其周围牙周组织观察组织学改变。与正畸牙齿移动相反，在复发过程中，近中腭根的远中侧牙周膜是压力区，近中侧是张力区(图1)。牙周膜中骨保护素染色的图像分析结果：对照组与实验组比较，压力区和张力区灰度积分，高、中、低剂量组与对照组比较，差异有显著性意义(P < 0.05)，实验组不同剂量相比，高剂量比中剂量组和低剂量组明显增加，差异有显著性意义(P < 0.01)。实验组与对照组牙周膜骨保护素阳性反应灰度积分比较，配对t 检验结果显示张力区略高于压力区，差异无显著性意义(P > 0.05，表3)。牙周膜中破骨细胞分化因子染色的图像分析结果：实验组与对照组比较，压力区和张力区破骨细胞分化因子灰度积分[23]，差异均有显著性意义(P < 0.01)；实验组不同剂量组比较，高剂量比中剂量组和低剂量组明显减低，差异均有显著性意义(P < 0.001)。实验组与对照组牙周膜破骨细胞分化因子阳性反应灰度积分，配对t 检验结果显示压力区略高于张力区，差异无显著性意义(P > 0.05，表3)。牙周膜中骨保护素/破骨细胞分化因子比率分析结果：骨保护素/破骨细胞分化因子比率大于1，说明骨保护素相对过剩，破骨细胞的分化和激活受到抑制，促进骨形成；骨保护素/破骨细胞分化因子比率小于1，破骨细胞分化因子相对过剩，促进破骨细胞的分化和激活。由表3可见：对照组比率小于1，实验组牙周组织骨保护素/破骨细胞分化因子比率均大于1，高剂量组比率最大。"

References

[1] Simonet WS,Lacey DL,Dunstan CR,et al. Osteoprotegerin: anovelseereted Protein Involved in the regulation of bone density.Cell.1997;89(2):309-319.

[2] Franceca GoBi,Lorenz C, Hof Bauer,et al.The expression of osteoprotegerion and RANK ligand support of osteoclast for mation by stromal-osteoblast lingeage cells is developmentally regulated.Endocrinology. 2000;141(12): 4768-4776.

[3] Yeung DK ,Leung SW, Xu YC, et al. Puerar in, anisof lavonoidderived fromRadix Puerariae, Potentiatesendonthelium in dependentr elaxat ionviatheeyelie AMPPathway In Poreinec oronary artery. Eur J Pharmaeol. 2006;552(1-3):105-111.

[4] 黄生高,康祖铭,张建兴,等.旋转脉动磁场加速兔正畸牙移动的实验研究[J].中国现代医学杂志,2005,15(20):3085-3088

[5] 张丁,傅民魁,傅嘉.丹参对离体破骨细胞的影响[J].口腔正畸学, 1995,2(2):63-64

[6] 沈刚,陈荣敬,刘泓虎,等.复方灯盏花加速兔牙移动的运动学模型的建立与分析[J].口腔正畸学杂志,1998,5(1):31-33

[7] 丁寅,陈华,徐如生.中药丹参加速正畸牙齿移动的研究[J].口腔医学,1995,15(3):120-121

[8] 中华人民共和国卫生部药典委员会.中国药典[S].北京:人民卫生出版社1985:291-292

[9] 中国医学科学院药物研究所编著．中草药现代研究(Ⅱ) [M].北京医科大学中国协和医科大学联合出版,1996:472.

[10] 郑虎占,董泽宏,祭靖．中药现代研究与应用[M].2卷.北京:学苑出版社,1997,10．

[11] 史炜镔,符诗聪,杜宁,等.丹参有效部位对骨折愈合过程中胶原基因表达的影响[J]中国中西医结合杂志,2000:20(4):269-271.

[12] 林润台,王忠,刘立波,等.丹参促进下颌骨骨折愈合的超微结构研究[J].中华口医学杂志,1992:27(4):215-216.

[13] Liu YR, Qu SX, Maitz MF, et al. The effect of the major components of Salvia Miltiorrhiza Bunge on bone marrow cells. J Ethnopharmacol. 2007 ;111(3):573-583.

[14] 陈良娇,兰泽栋,陈建明. rhIL-1α与1,25-(OH)2D3联合作用对人牙周膜成纤维细胞表达RANKL和OPG的影响[J].口腔医学研究, 2012;28(4):316-320.

[15] Ren Y, Maltha JC, Kuijpers-Jagtman AM. The rat as a model for orthodontic tooth movement a critical review and a proposed solution.Eur J Orthod.2004;26(5):483-490.

[16] 黄艳,王旭霞,张君,等.正畸牙移动实验动物模型的建立[J].口腔医学,2007,27(2):64-65.

[17] Hagashi H, Konoo T, Yamaquchi K, Intermittent 8-hour a ctivation in orthodontic molar movement.Am J Orthod Dentofacial Orthop.2004;125(3):302-309.

[18] Ong CK, Walsh LJ, Harbrow D.et al.Orthodontic tooth movement in the prednisolone-treated rat.Angle Orthod. 2000;70(2):118-125.

[19] Xu X, Zhang S, Zhang L, et al.The Neuro Proteetion of Puerarina gainst eerebralisehemia 15 associatedwiththe Preventionofa pop tosisinrats. Planta Med.2005;71(7):585-91.

[20] Garat JA, Martin AE, Gordillo ME,et al.Effect of orthodontic forces on toot resorption in molars submitted to experimental periodomtitis.Acta Odontol Latinoam.2004;17(1-2):3-7.

[21] 金作林,丁寅,李潇．雌激素治疗骨质疏松大鼠正畸牙移动的实验[J]．中华口腔医学杂志,2001,35(1):55-57．

[22] 陈丽萍.骨保护素的有关研究[J].牙体牙髓牙周病学杂志,2007, 17(6):361.

[23] 刘宁,冯云霞,陈显久.不同正畸力对大鼠牙周组织压力侧 RANKL表达影响的研究 [J].中国实用口腔科杂志,2012,5(2): 104-108.

[24] Liedert A, Wagner L, Seefried L, et al.Estrogen receptor and Wnt signaling interact to regulate early gene expression in response to mechanical strain in osteoblastic cells.Biochem Biophys.2010;394(3):755-759.

[25] Orava S, Hulkko A, Koskinen S, et al. Stress fractures and bonehealth in track and field athletes. Sports Sci Med. 2000; 3(3):268-279.

[26] Orava S, Hulkko A, Koskinen S, et al. Stress fractures in athletes military recruits. An overview.Orthopade. 1995;24(5): 457-466.

[27] Lyritis GP, Georgoulas T, Zafeiris CP.Bone anabolic versus bone anticatabolic treatment of postme nopausal osteoporosis. Ann N Y Acad. 2010;1205:277-283.

[28] 陈雁南,裴帆,李晓智,等.局部注射骨保护素与二磷酸盐对大鼠正畸牙移动的影响[J].解放军医学杂志,2011, 36(11):1200-1202, 1206.

[29] 陈雁南,裴帆,李晓智,等.局部注射不同浓度骨保护素对大鼠正畸牙移动影响的研究[J].激光杂志,2011,5:89-90.

[30] Bateman TA,Dunstan CR,Ferguson VL,et al．Osteoprotegerin mitigates tail suspension-induced osteopenia. Bone.2000; 26(5):443-449．

[31] Bekker PJ, Holloway DL, Rasmussen AS,et al. A single-dose placebo-controlled study of AMG 162, a fully human monoclonal antibody to RANKL, in postmenopausal women. J Bone Miner Res.2004;19:1059-1066.

[32] Manolagas SC,Jilka RL.Bone marrow,cytokines,and bone remodeling,Emerging insights into the pathophysiology of osteoponosis. N Engl J Med. 1995;332(5):305-311.

[33] Quinn JM,Elliott J,Gilespie MT,et al．Endocrinology.1998；139(10):4424-4427．

[34] 孔祥鹤,牛银波,李宇华,等.OPG/RANK/RANKL系统与骨质疏松研究最新进展[J].生命科学研究,2011,15(7):80-85.

[35] Asuda H.Bone and bone related biot•heroical examinati(ms.Bone anti collagen related metabolites. Receptor actiator of NF-kappaB ligand(RANKL)．Clin Calcium. 2006;16(6):964-970.

[36] Shiotani A, Takami M, Itoh K, et al．Regulation of Osteoclast Differentiation and Function by Receptor Activator of NFkB Ligand and Osteoprotegerin．Anatomical Record.2002; 268: 137-146．

[37] SahTlon P.Loss of chaotic trabecular structure in OPG-deficient juvenile Paget's disease patients indicates a chaogenic role for OPG in nonlinear pattern formation of trabecular bone.J Bone Miner Res. 2004;19(5):695-702．

[38] 任嫒姝．应力刺激下成牙骨质细胞OPG/RANKL的表达及其信号转导通路的研究[D]．成都:四川大学华西口腔医学院,2007．

[39] Tang L, Lin Z, Li YM．Effects of different magnitudes of mechanical strain on Osteoblasts in vitro. Biochem Biophys Res Commun. 2006;344(1):122-128.

[40] 贾少杰,贾晓静．绝经后女性骨质疏松与OPG、RANKL和ApoE的相关性研究[J]．中国妇幼保健,2007,32(3):1001-1441．

[41] 高延征,高坤,沈彬,等.去卵巢后大鼠骨组织中核因子κB受体活化因子配体和骨保护素表达的变化[J].中国组织工程研究与临床康复,2009,13(15):2877-2881．

[42] 梁少俊,刘宏,杨力.绝经后骨质疏松与血清OPG及RANKL关系的研究[J].广东医学,2006,27(5):669-671．

[43] 祝金香,张纲,武曦,等.RANKL/OPG 在高原低氧条件下兔牙周炎发病机制中的作用[J].第三军医大学学报,2012,(34)4: 320-323

[44] Itonaga I,Fujikawa Y,Sabokbar A,et al.Rheumatoid arthritis synovial macrophage-osteoclast differentiation is osteoprotegerin ligand-dependent.J Pathol. 2000;192(1): 97-104.

[45] Teng YT, Nguyen H,Kong YY, et al.Functional human T-cell immunity and osteoprotegerin ligand control alveolar bone destruction in periodontal infection. J Clin Invest.2000;

106(6):59-67.

[46] Mogi M, Otogoto J, Ota N, et al.Differential expression of RANKL and osteoprotegerin in gingival crevicular fluid of patients with periodontitis. J Dent Res. 2004;83(2):166-169.