降钙素基因相关肽对人脐带间充质干细胞成骨分化的影响

高振; 吴琴; 郭俊峰; 李铀; 陈雪丹; 高强国; 张纲

doi:10.16016/j.1000-5404.201711055

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降钙素基因相关肽对人脐带间充质干细胞成骨分化的影响

基础医学 | 更新时间：2026-01-09

- 降钙素基因相关肽对人脐带间充质干细胞成骨分化的影响
- Effects of calcitonin gene-related peptide on osteogenic differentiation of human umbilical cord mesenchymal stem cells
- 第三军医大学学报 2018年40卷第6期页码：479-486
- 作者机构：
  
  1.400037 重庆，陆军军医大学(第三军医大学)第二附属医院口腔科
  2.400038 重庆，陆军军医大学(第三军医大学)基础医学院医学遗传学教研室
- 作者简介：
  
  [ 张纲, E-mail: xqyykqk@163.com ZHANG Gang, E-mail: xqyykqk@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81271098)
- DOI：10.16016/j.1000-5404.201711055
  中图分类号：
- 收稿：2017-11-08，
  
  修回：2017-12-19，
  
  纸质出版：2018-03-30
- 稿件说明：
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高振, 吴琴, 郭俊峰, 等. 降钙素基因相关肽对人脐带间充质干细胞成骨分化的影响[J]. 第三军医大学学报, 2018,40(6):479-486.

Zhen GAO, Qin WU, Junfeng GUO, et al. Effects of calcitonin gene-related peptide on osteogenic differentiation of human umbilical cord mesenchymal stem cells[J]. Journal of Third Military Medical University, 2018, 40(6): 479-486.
高振, 吴琴, 郭俊峰, 等. 降钙素基因相关肽对人脐带间充质干细胞成骨分化的影响[J]. 第三军医大学学报, 2018,40(6):479-486. DOI： 10.16016/j.1000-5404.201711055.

Zhen GAO, Qin WU, Junfeng GUO, et al. Effects of calcitonin gene-related peptide on osteogenic differentiation of human umbilical cord mesenchymal stem cells[J]. Journal of Third Military Medical University, 2018, 40(6): 479-486. DOI： 10.16016/j.1000-5404.201711055.

摘要

目的

探讨降钙素基因相关肽(calcitonin gene related peptide，CGRP)在人脐带间充质干细胞(human umbilical cord mesenchymalstem cells，HUMSCs)向成骨细胞分化过程中的作用及机制。

方法

体外培养HUMSCs细胞，加入0、10

-9

、10

-8

、10

-7

mol/L浓度的CGRP诱导其向成骨细胞分化，分别在第7、9、11、14天时采用茜素红染色检测成骨分化程度，实时定量PCR和Western blot检测ERK、RUNX2、OSTERIX的mRNA和磷酸化的ERK、RUNX2、OSTERIX的蛋白表达。

结果

HUMSCs高表达CD73、CD90、CD105等间充质干细胞表面抗原，但不表达CD14、CD20、CD34、CD45等上皮和造血干细胞的表面抗原，可诱导向成骨细胞分化，随着时间延长矿化结节数量增加(

＜0.05)；随着CGRP浓度的升高，矿化结节减少，ERK、RUNX2、OSTERIX的mRNA和蛋白表达降低(

＜0.05)。

结论

CGRP通过ERK信号通路抑制HUMSCs向成骨细胞的分化，并呈现出浓度和时间依赖性。

Abstract

Objective

To investigate the role and underlying mechanism of calcitonin gene-related peptide (CGRP) in the differentiation of human umbilical cord mesenchymal stem cells (HUMSCs) into osteoblasts.

Methods

Different concentrations (0

-9

-8

and 10

-7

mol/L) of CGRP were added into the culture media of HUMSCs to induce differentiation into osteoblasts. In 7

11 and 14 d after treatment

Alizarin red staining was used to detect the differentiation of osteogenesis. Real-time PCR and Western blotting were employed to detect the mRNA expression levels of ERK

RUNX2 and OSTERIX and protein expression levels of phosphorylation of ERK

RUNX2 and OSTERIX.

Results

HUMSCs with high expression of CD73

CD90

CD105 and other mesenchymal stem cell surface antigens

but not with the expression of CD14

CD20

CD34

CD45 and other epithelial and hematopoietic stem cell surface antigens

were identified to induce differentiation into osteoblasts. The number of mineralized nodules was increased with time elapsing (

＜0.05)

while the number was decreased with the increasing concentrations of CGRP

and the mRNA and protein expression levels of ERK

RUNX2 and OSTERIX were decreased (

＜0.05).

Conclusion

CGRP inhibits the differentiation of HUMSCs into osteoblasts by ERK signaling pathway in a concentration-and time-dependent manner.

关键词

Keywords

references

常正奇, 许建中, 梁进, 等.人脐带间充质干细胞的新分离方法及其作为骨组织工程种子细胞的相关研究[J].第三军医大学学报, 2015, 37(19): 1930-1935. DOI: 10.16016/j.1000-5404.201501035.

CHANG Z Q, XU J Z, LIANG J, et al .New isolation method of human umbilical cord mesenchymal stem cells and relative research of them as seed cells in tissue engineering bone[J ] . J Third Mil Med Univ, 2015, 37(19): 1930-1935. DOI: 10.16016/j.1000-5404.201501035.

LOH Y, OYAMA Y, STATKUTE L, et al . Development of a secondary autoimmune disorder after hematopoietic stem cell transplantation for autoimmune diseases: role of conditioning regimenused[J ] . Blood, 2007, 109(6): 2643-2648. DOI: 10.1182/blood-2006-07-035766.

AGGARWAL S, PITTENGER M F. Human mesenchymal stem cells modulate allogeneic immune cell responses[J]. Blood, 2005, 105(4): 1815-1822. DOI: 10.1182/blood-2004-04-1559.

SAMPLE S J, HAO Z, WILSON A P, et al . Role of calcitonin gene-related peptide in bone repair after cyclic fatigue loading[J ] . PLoS ONE, 2011, 6(6): e20386. DOI: 10.1371/journal.pone.0020386.

ZHAO X G, ZHAO G F, MA Y F, et al . Research progress in mechanism of traumatic brain injury affecting speed of fracture healing[J ] . Chin J Traumatol, 2007, 10(6): 376-380.

DA PAZ A C, CAROD ARTAL F J, KALIL R K. The function of proprioceptors in bone organization: a possible explanation for neurogenic heterotopic ossification in patients with neurological damage[J]. Med Hypotheses, 2007, 68(1): 67-73. DOI: 10.1016/j.mehy.2006.06.035.

IMAI S, MATSUSUE Y. Neuronal regulation of bone metabolism and anabolism: calcitonin gene-related peptide-, substance P-, and tyrosine hydroxylase-containing nerves and the bone[J]. Microsc Res Tech, 2002, 58(2): 61-69. DOI: 10.1002/jemt.10119.

LI J, KREICBERGS A, BERGSTROM J, et al . Site-specific CGRP innervation coincides with bone formation during fracture healing and modeling: a study in rat angulated tibia[J ] . J Orthop Res, 2007, 25(9): 1204-1212. DOI: 10.1002/jor.20406.

VILLA I, MRAK E, RUBINACCI A, et al . CGRP inhibits osteoprotegerin production in human osteoblast-like cells via cAMP/PKA-dependent pathway[J ] . Am J Physiol Cell Physiol, 2006, 291(3): C529-C537. DOI: 10.1152/ajpcell.00354.2005..

MRAK E, GUIDOBONO F, MORO G, et al . Calcitonin gene-related peptide (CGRP) inhibits apoptosis in human osteoblasts by beta-catenin stabilization[J ] . J Cell Physiol, 2010, 225(3): 701-708. DOI: 10.1002/jcp.22266.

ZHANG Y, YANG J, ZHANG P, et al . Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury[J ] . Sci Rep, 2016, 6: 27724. DOI: 10.1038/srep27724.

MARUPANTHORN K, TANTRAWATPANC, KHEOLAMAI P, et al . Bone morphogenetic protein-2 enhances the osteogenic differentiation capacity of mesenchymal stromal cells derived from human bone marrow and umbilical cord[J ] . Int J Mol Med, 2017, 39( 3): 654-662. DOI: 10.3892/ijmm.2017.2872.

KIM J, SHIN J M, JEON Y J, et al . Proteomic validation of multifunctional molecules in mesenchymal stem cells derived from human bone marrow, umbilical cord blood and peripheral blood[J ] . PLoS ONE, 2012, 7(5): e32350. DOI: 10.1371/journal.pone.0032350.

HEO J S, CHOI Y, KIM H S, et al . Comparison of molecular profiles of human mesenchymal stem cells derived from bone marrow, umbilical cord blood, placenta and adipose tissue[J ] . Int J Mol Med, 2016, 37(1): 115-125. DOI: 10.3892/ijmm.2015.2413.

LEE J K, JUNG J S, PARK S H, et al . Deficiency of alpha-calcitonin gene-related peptide induces inflammatory responses and lethality in sepsis[J ] . Cytokine, 2013, 64(2): 548-554. DOI: 10.1016/j.cyto.2013.07.030.

LONG H, SUN B, CHENG L, et al . miR-139-5p Represses BMSC osteogenesis via targeting Wnt/beta-Catenin signaling pathway[J ] . DNA Cell Biol, 2017, 36(8): 715-724. DOI: 10.1089/dna.2017.3657..

FAKHRY M, HAMADE E, BADRANB, et al . Molecular mechanisms of mesenchymal stem cell differentia tion towards osteoblasts[J ] . World J Stem Cells, 2013, 5(4): 136-148. DOI: 10.4252/wjsc.v5.i4.136.

MURAKAMI J, ISHⅡ M, SUEHIRO F, et al . Vascular endothelial growth factor-C induces osteogenic differentiation of human mesenchymal stem cells through the ERK and RUNX2 pathway[J ] . Biochem Biophys Res Commun, 2017, 484(3): 710-718. DOI: 10.1016/j.bbrc.2017.02.001.

LI B, QU C, CHEN C, et al . Basic fibroblast growth factor inhibits osteogenic differentiation of stem cells from human exfoliated deciduous teeth through ERK signaling[J ] . Oral Dis, 2012, 18(3): 285-292. DOI: 10.1111/j.1601-0825.2011.01878.x.

ZHAO Y, ZHANG S, ZENG D, et al . rhPDGF-BB promotes proliferation and osteogenic differentiation of bone marrow stromal cells from streptozotocin-induced diabetic rats through ERK pathway[J ] . Biomed Res Int, 2014, 2014: 637415. DOI: 10.1155/2014/637415.

邹薇, 刘慧通, 焦富勇.ERK信号传导通路在人脐带间充质干细胞向成骨细胞分化作用中的研究进展[J].中国美容医学, 2010, 19(10): 1569-1571. DOI: 10.3969/j.issn.1008-6455.2010.10.069.

ZOU W, LIU H T, JIAO F Y. Research associated with the role of the ERK signaling pathway which played in osteogenic differentiation of the human umbilical cord mesenchymal stem cells[J]. Chin J Aesthetic Med, 2010, 19(10): 1569-1571. DOI: 10.3969/j.issn.1008-6455.2010.10.069.

KIM J M, LEE J E, RYU S H, et al . Chlormadinone acetate promotes osteoblast differentiation of human mesenchymal stem cells through the ERK signaling pathway[J ] . Eur J Pharmacol, 2014, 726: 1-8. DOI: 10.1016/j.ejphar.2014.01.013.

ONO T, TAKAYANAGI H. Osteoimmunology in Bone Fracture Healing[J]. Curr Osteoporos Rep, 2017, 15(4): 367-375. DOI: 10.1007/s11914-017-0381-0.

FIERRO F A, NOLTA J A, ADAMOPOULOS I E. Concise review: stem cells in osteoimmunology[J]. Stem Cells, 2017, 35(6): 1461-1467. DOI: 10.1002/stem.2625.

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