中国医科大学学报

中国医科大学学报
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中国医科大学学报 ›› 2018, Vol. 47 ›› Issue (7): 617-621.doi: 10.12007/j.issn.0258-4646.2018.07.011

• 论著 • 上一篇    下一篇

Wnt3a对人骨髓间充质干细胞成骨分化的影响及机制研究

赵芳英, 高秀秋, 刘姊凤   

  1. 锦州医科大学附属第二医院牙体牙髓科, 辽宁 锦州 121000
  • 收稿日期:2017-11-16 出版日期:2018-07-30 发布日期:2018-07-03
  • 通讯作者: 赵芳英 E-mail:229157364@qq.com
  • 作者简介:赵芳英(1970-),女,副主任医师,硕士.
  • 基金资助:
    辽宁省科学事业公益研究基金(2015001014)

Effect and Mechanism of Wnt3a on Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells

ZHAO Fangying, GAO Xiuqiu, LIU Zifeng   

  1. Department of Endodontics, The Second Affiliated Hospital, Jinzhou Medical University, Jinzhou 121000, China
  • Received:2017-11-16 Online:2018-07-30 Published:2018-07-03

摘要: 目的 探讨Wnt3a对人骨髓间充质干细胞成骨分化及Wnt/β-catenin信号通路的影响。方法 采用成骨诱导培养基联合Wnt3a蛋白(10 ng/mL)诱导人骨髓间充质干细胞分化,通过镜下形态学和茜素红染色评价成骨分化水平。采用实时PCR及Western blotting法检测Wnt3a干预下骨髓间充质干细胞分化过程中第1、5、10、15和20天RUNX2、β-catenin及BSP的表达情况。结果Wnt3a干预下充质干细胞向成骨细胞分化程度明显高于对照组。Wnt3a能提高RUNX2、β-catenin及BSP mRNA和蛋白的表达水平(P < 0.05),表达于干预第10天时达到高峰,随后呈轻度下降趋势。结论 Wnt3a通过激活Wnt/β-catenin信号通路调控人骨髓间充质干细胞成骨分化。

关键词: 人骨髓间充质干细胞, 成骨分化, Wnt3a, β-catenin, RUNX2, BSP

Abstract: Objective To investigate the effect of Wnt3a on the osteogenic differentiation of human bone marrow mesenchymal stem cells and the Wnt/β-catenin signaling pathway. Methods We induced the differentiation of human bone marrow mesenchymal stem cells using an osteogenic induction medium supplemented with the Wnt3a protein (10 ng/mL) to evaluate its osteogenic differentiation by microscopic morphology and alizarin red staining. Real-time PCR and Western blotting were used to detect the expression of Runt-related transcription factor 2 (RUNX2), β-catenin, and bone sialoprotein (BSP) on day 1, 5, 10, 15, and 20 after treatment with Wnt3a. Results Mesenchymal stem cell differentiation into osteoblasts was significantly increased by Wnt3a compared with that of the control group. Moreover, Wnt3a increased the expression of RUNX2, β-catenin, and BSP (P < 0.05), levels of which reached a peak at day 10 after treatment with Wnt3a. Conclusion Wnt3a participated in and regulated the osteogenic differentiation of human bone marrow mesenchymal stem cells by activating the Wnt/β-catenin signaling pathway.

Key words: bone marrow mesenchymal stem cells, osteogenic differentiation, Wnt3a, β-catenin, RUNX2, BSP

中图分类号: 

  • R780.2
[1] OHISHI M,SCHIPANI E. Bone marrow mesenchymal stem cells[J]. J Cell Biochem,2010,109(2):277-282. DOI:10.1002/jcb.22399.
[2] CAWTHORN WP,BREE AJ,YAO Y,et al. Wnt6,Wnt10a and Wnt10b inhibit adipogenesis and stimulate osteoblastogenesis through β-catenin-dependent mechanism[J]. Bone,2012,50(2):477-489. DOI:10.1016/j.bone.2011.08.010.
[3] VELASCO J,ZARRABEITIA MT,PRIETO JR,et al. Wnt pathway genes in osteoporosis and osteoarthritis:differential expression and genetic association study[J]. Osteoporos Int,2010,21(1):109-118. DOI:10.1007/s00198-009-0931-0.
[4] XU ML,BI CWC,LIU EYL,et al. Wnt3a induces the expression of acetylcholinesterase during osteoblast differentiation via the Runx2 transcription factor[J]. J Biol Chem,2017,292(30):12667-12678. DOI:10.1074/jbc.M117.777581.
[5] GILCHRIST ES,PLEVRIS JN. Bone marrow-derived stem cells in liver repair:10 years down the line[J]. Liver Transpl,2011,16(2):118-129. DOI:10.1002/lt.21965.
[6] ZHANG Z,WANG X,ZHANG L,et al. Wnt/β-catenin signaling pathway in trophoblasts and abnormal activation in preeclampsia[J]. Mol Med Rep,2017,16(2):1007-1013. DOI:10.3892/mmr.2017.6718.
[7] BOLAND M,PERKINS G,HALL DJ,et al. Wnt 3a promotes proliferation and suppresses osteogenic differentiation of adult human mesenchymal stem cells[J]. J Cell Biochem,2004,93(6):1210-1230. DOI:10.1002/jcb.20284.
[8] LI YS,XI Y,LI XJ,et al. Up-regulation of the biosynthesis and release of substance p through wnt/β-catenin signaling pathway in rat dorsal root ganglion cells[J]. PLoS One,2015,10(6):e0129701. DOI:10.1371/journal.pone.0129701.
[9] DENYSENKO T,ANNOVAZZI L,CASSONI P,et al. WNT/β-catenin signaling pathway and downstream modulators in low-and high-grade glioma[J]. Cancer Genomics Proteomics,2016,13(1):31-45.
[10] 周大凯,李慧宁,马珊珊,等. 基因高表达对脐血间充质干细胞成骨分化相关基因表达的影响[J]. 中国组织工程研究,2017,21(9):1444-1449. DOI:10.3969/j.issn.2095-4344.2017.09.024
[11] 涂艳,熊莉娜,柳湘洁,等. 淫羊藿苷对成骨细胞成骨分化的影响及Wnt/β-catenin信号系统的关系研究[J]. 中国中医急症, 2017,26(3):448-450. DOI:10.3969/j.issn.1004-745X.2017.03. 023.
[12] WANG Y,UEMURA T,DONG J,et al. Application of perfusion culture system improves in vitro and in vivo osteogenesis of bone marrow-derived osteoblastic cells in porous ceramic materials[J]. Tissue Eng,2003,9(6):1205-1214. DOI:10.1089/10763270360728116.
[13] CHO HH,KIM YJ,KIM SJ,et al. Endogenous Wnt signaling promotes proliferation and suppresses osteogenic differentiation in human adipose derived stromal cells[J]. Tissue Eng,2006,12(1):111-121. DOI:10.1089/ten.2006.12.111.
[14] DE BOER J,SIDDAPPA R,GASPAR C,et al. Wnt signaling inhibits osteogenic differentiation of human mesenchymal stem cells[J]. Bone,2004,34(5):818-826. DOI:10.1016/j.bone.2004.01.016.
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