Wnt5a在小鼠牙骨质发育中的表达特点及其对成牙骨质细胞分化的影响

doi:10.13481/j.1671-587x.20180301

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (03): 461-465.doi: 10.13481/j.1671-587x.20180301

• 基础研究 • 下一篇

Wnt5a在小鼠牙骨质发育中的表达特点及其对成牙骨质细胞分化的影响

包幸福¹, 倪宇昕², 李莹¹, 郭小凯¹, 王玉琢¹, 方蛟², 胡敏¹

1. 吉林大学口腔医院正畸科, 吉林长春 130021;
2. 吉林大学口腔医院种植科, 吉林长春 130021

收稿日期:2017-09-12 出版日期:2018-05-28 发布日期:2018-05-31
通讯作者: 胡敏,教授,博士研究生导师(Tel:0431-88796023,E-mail:humin@jlu.edu.cn) E-mail:humin@jlu.edu.cn
作者简介:包幸福(1986-),男,吉林省长春市人,主治医师,医学博士,从事牙齿硬组织发生和改建方面的研究。
基金资助:
国家自然科学基金资助课题（81470764）；吉林省卫计委青年科研基金资助课题（2015Q003，2015Q013）；吉林大学白求恩计划项目资助课题（2015321）；吉林省科技厅科技发展计划项目资助课题（20160520156JH）

Expression characteristics of Wnt5a in different periods of cementum development and its effect on differentiationof cementoblasts of mice

BAO Xingfu¹, NI Yuxin², LI Ying¹, GUO Xiaokai¹, WANG Yuzhuo¹, FANG Jiao², HU Min¹

1. Department of Orthodontics, Stomatology Hospital, Jilin University, Changchun 130021, China;
2. Department of Implantation, Stomatology Hospital, Jilin University, Changchun 130021, China

Received:2017-09-12 Online:2018-05-28 Published:2018-05-31

摘要/Abstract

摘要： 目的：探讨Wnt家族5A蛋白（Wnt5a）在小鼠出生后牙骨质中的表达特点以及其对体外培养成牙骨质细胞分化的影响，阐明Wnt非经典通路调控牙根发育的机制。方法：选择出生后0.5、4.5、10.5、16.5、20.5、24.5、26.5和30.5d昆明小鼠并按以上相应时间点分组，每组3只；在相应时间点采用脱臼法处死小鼠，分离下颌第一磨牙及牙周组织。免疫组织化学染色法观察小鼠牙体和牙周组织中Wnt5a的表达特点。将小鼠成牙骨质细胞OCCM30分为实验组和对照组。实验组加入200μg·L^-1重组Wnt5a蛋白，对照组无添加；提取细胞总RNA。RT-PCR法检测2组OCCM30细胞中Ocn、Opn、Alp、Bsp、Wnt5a和Runx-2基因的相对表达水平。结果：小鼠出生后0.5d，第一磨牙牙胚中无Wnt5a阳性表达；出生后4.5d，成釉细胞和成牙本质细胞中Wnt5a阳性表达较强；出生后16.5d，可见成牙骨质细胞中Wnt5a呈阳性表达；出生后20.5~30.5 d，成牙本质细胞和牙周膜细胞中Wnt5a呈阳性表达。与对照组比较，实验组成牙骨质细胞中Ocn和Alp基因相对表达水平降低（P<0.05）。结论：Wnt5a在牙根发育过程中的表达特点为从无到有，呈现规律性表达；Wnt5a下调分化基因的表达，在牙骨质发育中发挥重要作用。

关键词: Wnt5a, 牙骨质, 免疫组织化学, 牙发育, 成牙骨质细胞

Abstract: Objective: To study the expression characteristics of Wnt5a in cementum of the mice after birth and the effect of Wnt5a on the differentiation of cementoblasts in vitro,and to illuminate the mechanism of Wnt noncanonical signaling in regulating the tooth development. Methods: The postnatal Kunming mice at the days 0.5,4.5,12.5,18.5,24.5 and 30.5 after birth were selected and divided into various groups by different time points (n=3).The mice were sacrificed and the lower first molar and peridentium were prepared.Immunohistochemical staining was used to detect the expression characteristics of Wnt5a in tooth and peridentium of the mice.The OCCM30 cells were divided into control group and experiment group.200 μg·L^-1 recombinant Wnt5a protein were added in experiment group, and nothing was added in control group;the total RNA in the cells was exrracted.The relative expression levels of Ocn,Opn,Alp,Bsp,Wnt5a,and Runx-2 genes in the OCCM30 cells in two groups were detected by RT-PCR method. Results: At the day 0.5 after birth,the expression of Wnt5a in the lower first molar tooth germ was negative.At the day 4.5 after birth,the expression of Wnt5a in ameloblasts and odontoblasts were positive.At the day 16.5 after birth,the expression of Wnt5a in cementoblasts was positive.At the days 20.5-30.5 after birth,the expressions of Wnt5a in odontoblasts and periodontal ligament cells were positive.Compared with control group,the relative expression levels of Ocn and Alp gene in the OCCM30 cells in experiment group were decreased (P<0.05). Conclusion: The expression of Wnt5a is regular at different tooth developmental stages after birth.Wnt5a can reduce the expression of differentiated genes and play an important role in the growth of cementum.

Key words: tooth development, Wnt5a, cementoblasts, immunohistochemistry, cementum

中图分类号:

包幸福, 倪宇昕, 李莹, 郭小凯, 王玉琢, 方蛟, 胡敏. Wnt5a在小鼠牙骨质发育中的表达特点及其对成牙骨质细胞分化的影响[J]. 吉林大学学报(医学版), 2018, 44(03): 461-465.

BAO Xingfu, NI Yuxin, LI Ying, GUO Xiaokai, WANG Yuzhuo, FANG Jiao, HU Min. Expression characteristics of Wnt5a in different periods of cementum development and its effect on differentiationof cementoblasts of mice[J]. Journal of Jilin University Medicine Edition, 2018, 44(03): 461-465.

参考文献

[1] Foster BL.On the discovery of cementum[J].J Periodont Res,2017,52 (4):666-685.
[2] Yamamoto T,Hasegawa T,Yamamoto T,et al.Histology of human cementum:Its structure,function,and development[J].Jpn Dent Sci Rev,2016,52 (3):63-74.
[3] Li J,Parada C,Chai Y.Cellular and molecular mechanisms of tooth root development[J].Development,2017,144 (3):374-384.
[4] Tamura M,Nemoto E.Role of the Wnt signaling molecules in the tooth[J].Jpn Dent Sci Rev,2016,52 (4):75-83.
[5] Peng L,Ye L,Dong G,et al.WNT5A inhibits human dental papilla cell proliferation and migration[J].Biochem Biophys Res Commun,2009,390 (3):1072-1078.
[6] Iglesias-Linares A,Hartsfield JK Jr.Cellular and molecular pathways leading to external root resorption[J].J Dent Res,2017,96 (2):145-152.
[7] Kinane DF,Stathopoulou PG,Papapanou PN.Periodontal diseases[J].Nat Rev Dis Primers,2017,3:17038.
[8] Li J,Feng J,Liu Y,et al.BMP-SHH signaling network controls epithelial stem cell fate via regulation of its niche in the developing tooth[J].Dev Cell,2015,33 (2):125-135.
[9] Liu Y,Feng J,Li J,et al.An Nfic-hedgehog signaling cascade regulates tooth root development[J].Development,2015,142 (19):3374-3382.
[10] Kim TH,Bae CH,Lee JC,et al.β-catenin is required in odontoblasts for tooth root formation[J].J Dent Res,2013,92 (3):215-221.
[11] Han X L,Liu M,Voisey A,et al.Post-natal effect of overexpressed DKK1 on mandibular molar formation[J].J Dent Res,2011,90 (11):1312-1317.
[12] Kumawat K,Gosens R.WNT-5A signaling and functions in health and disease[J].Cell Mol Life Sci,2016,73 (3):567-587.
[13] Zhou Y,Kipps TJ,Zhang S.Wnt5a signaling in normal and cancer stem cells[J].Stem Cells Int,2017:5295286.
[14] Gibson JD,O'Sullivan MB,Alaee F,et al.Regeneration of articular cartilage by human esc-derived mesenchymal progenitors treated sequentially with BMP-2 and Wnt5a[J].Stem Cells Transl Med,2017,6(1):40-50.
[15] Kobayashi Y,Uehara S,Udagawa N,et al.Regulation of bone metabolism by Wnt signals[J].J Biochem,2016,159 (4):387-392.
[16] Maeda K,Kobayashi Y,Udagawa N,et al.Wnt5a-Ror2 signaling between osteoblast-lineage cells and osteoclast precursors enhances osteoclastogenesis[J].Nat Med,2012,18(3):405-412.
[17] Stricker S,Rauschenberger V,Schambony A.ROR-family receptor tyrosine kinases[J].Curr Top Dev Biol,2017,123:105-142.
[18] Peng L,Ye L,Dong G,et al.WNT5A inhibits human dental papilla cell proliferation and migration[J].Biochem Biophys Res Commun,2009,390 (3):1072-1078.
[19] Sakisaka Y,Tsuchiya M,Nakamura T,et al.Wnt5a attenuates Wnt3a-induced alkaline phosphatase expression in dental follicle cells[J].Exp Cell Res,2015,336 (1):85-93.
[20] Wang YL,He H,Liu ZJ,et al.Effects of TNF-α on cementoblast differentiation,mineralization,and apoptosis[J].J Dent Res,2015,94(9):1225-1232.

Wnt5a在小鼠牙骨质发育中的表达特点及其对成牙骨质细胞分化的影响

Expression characteristics of Wnt5a in different periods of cementum development and its effect on differentiationof cementoblasts of mice

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 10

[1]	李红霞, 宫海叶, 曾萧, 张素素, 刘伯锋. TGF-β1和PLGF在不同类型妊娠滋养细胞疾病患者病变组织中的表达及其意义[J]. 吉林大学学报(医学版), 2018, 44(05): 1041-1046.
[2]	赵振理, 畅继武, 孙光, 胡少华. AAX14401蛋白在人膀胱癌组织中的表达及其意义[J]. 吉林大学学报(医学版), 2018, 44(05): 1052-1055.
[3]	王天月, 崔晓燕, 吴骁, 王丹. 磷酸化胞外信号调节激酶在小鼠海马发育过程中的表达及其意义[J]. 吉林大学学报(医学版), 2018, 44(05): 979-982.
[4]	阎慧, 娄石磊, 苗永迪, 岂蕊, 郭子硕, 邱悦, 孙聪. 大鼠结肠原代成纤维细胞的分离、培养和鉴定[J]. 吉林大学学报(医学版), 2018, 44(04): 849-852.
[5]	张啸环, 段明华, 闫玉礼, 潘新, 李海清, 周长龙, 赵天倚. 人羊膜成纤维细胞的分离和鉴定[J]. 吉林大学学报(医学版), 2018, 44(04): 845-848.
[6]	杨易, 包康达, 周彦兵, 袁超, 李勇, 刘晓明, 徐金瑞. Wnt5a对人肺腺癌A549细胞凋亡的调控作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(02): 229-234.
[7]	马晓周, 刘麒麟, 尹唯凰, 王越, 于阳, 杨刚, 于子莹. CC趋化因子受体6在人唾液腺腺样囊性癌组织中的表达及其临床意义[J]. 吉林大学学报(医学版), 2018, 44(01): 147-150.
[8]	曹艳莎, 李婳, 陈明红, 刘宝琴, 王华芹, 李宁. 组织芯片技术检测BAG3蛋白在结肠癌组织中的表达及其临床意义[J]. 吉林大学学报(医学版), 2017, 43(06): 1177-1181.
[9]	高新雅, 张伟杰, 崔黎, 周学良, 马志俊, 吕转, 池艳艳, 王留兴. TM4SF1在乳腺癌组织中的表达及其临床意义[J]. 吉林大学学报(医学版), 2017, 43(06): 1186-1192.
[10]	周世平, 徐红, 王琇, 李蓥. 内镜黏膜下剥离术治疗食管颗粒细胞瘤2例报告并文献复习[J]. 吉林大学学报(医学版), 2017, 43(05): 1034-1036.
[11]	历春, 付天悦, 杨宝学, 盖晓东. 尿素通道蛋白B在人膀胱癌组织中的表达及其临床意义[J]. 吉林大学学报(医学版), 2017, 43(04): 705-708.
[12]	石俊忠, 庄建彬, 张传山, 刘贤明, 孔大陆. β-tubulin Ⅲ在结肠腺癌组织中的表达及其临床意义[J]. 吉林大学学报(医学版), 2017, 43(04): 770-775.
[13]	郭小凯, 包幸福, 杨军星, 王玉琢, 李雨桐, 胡敏. 出生后小鼠牙不同发育期DKK1表达的形态学特点及其意义[J]. 吉林大学学报(医学版), 2017, 43(02): 241-244.
[14]	文爱平, 汪海娇, 金景姬, 蔡勇, 朱继红, 谌伦华. 子宫内膜癌组织中hMOF蛋白和mRNA表达及其临床意义[J]. 吉林大学学报(医学版), 2016, 42(06): 1138-1142.
[15]	温泉, 王园园, 司鹤南, 田亚萍. 成人郎格汉斯细胞组织细胞增生症1例报告及文献复习[J]. 吉林大学学报(医学版), 2016, 42(05): 985-987.