Determination of the minimal melatonin exposure required to induce osteoblast differentiation from human mesenchymal stem cells and these effects on downstream signaling pathways

被引:106
|
作者
Sethi, Shalini [1 ]
Radio, Nicholas M. [2 ]
Kotlarczyk, Mary P. [1 ]
Chen, Chien-Tsun [3 ,4 ]
Wei, Yau-Huei [3 ,4 ]
Jockers, Ralf [5 ,6 ]
Witt-Enderby, Paula A. [1 ]
机构
[1] Duquesne Univ, Sch Pharm, Div Pharmaceut Sci, Pittsburgh, PA 15282 USA
[2] Thermo Fisher Sci, Pittsburgh, PA USA
[3] Natl Yang Ming Univ, Dept Biochem & Mol Biol, Taipei 112, Taiwan
[4] MacKay Med Coll, Dept Med, Taipei, Taiwan
[5] Univ Paris 05, Inst Cochin, CNRS, UMR 8104, Paris, France
[6] INSERM, U1016, Paris, France
关键词
beta-arrestin scaffolds; MEK; ERK1; 2; mesenchymal stem cells; MT2 melatonin receptors; osteoblasts; BONE MORPHOGENETIC PROTEIN-2; HAMSTER OVARY CELLS; GROWTH-FACTOR-BETA; IN-VITRO; OSTEOGENIC DIFFERENTIATION; GENE-EXPRESSION; ALKALINE-PHOSPHATASE; PARATHYROID-HORMONE; POSTMENOPAUSAL OSTEOPOROSIS; REACTIVE OXYGEN;
D O I
10.1111/j.1600-079X.2010.00784.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The purpose of this study was to determine the critical time periods of melatonin treatment required to induce human mesenchymal stem cells (hAMSCs) into osteoblasts and to determine which osteogenic genes are involved in the process. The study design consisted of adding melatonin for different times (2, 5, 10, 14 or 21 days) toward the end of a 21-day treatment containing osteogenic (OS+) medium or at the beginning of the 21-day treatment and then withdrawn. The results show that a 21-day continuous melatonin treatment was required to induce both alkaline phosphatase (ALP) activity and calcium deposition and these effects were mediated through MT(2)Rs. Functional analysis revealed that peak ALP levels induced by melatonin were accompanied by attenuation of melatonin-mediated inhibition of forskolin-induced cAMP accumulation. Immunoprecipitation and western blot analyses, respectively, showed that MT2R/beta-arrestin scaffolds complexed to Gi, MEK1/2 and ERK1/2 formed in these differentiated hAMSCs (i.e., when ALP levels were highest) where ERK1/2 resided primarily in the cytosol. It is hypothesized that these complexes form to modulate the subcellular localization of ERK1/2 to affect osteogenic gene expression. Using real-time RT-PCR, chronic melatonin exposure induced the expression of osteogenic genes RUNX-2, osteocalcin and BMP-2, through MT(2)Rs. No melatonin-mediated changes in the mRNA expression of ALP, BMP-6 or in the oxidative enzymes MtTFA, PGC-1 alpha, Pol gamma, NRF-1, PDH, PDK and LDH occurred. These data show that a continuous 21-day melatonin exposure is required to induce osteoblast differentiation from hAMSCs through the formation of MT2R/Gi/beta-arrestin/MEK/ERK1/2 complexes to induce osteogenesis.
引用
收藏
页码:222 / 238
页数:17
相关论文
共 50 条
  • [21] Notch signaling in chondrogenic differentiation of human mesenchymal stem cells
    Hiraoka, K
    Lotz, M
    OSTEOARTHRITIS AND CARTILAGE, 2004, 12 : S103 - S104
  • [22] Skeletal (stromal) stem cells: An update on intracellular signaling pathways controlling osteoblast differentiation
    Abdallah, Basem M.
    Jafari, Abbas
    Zaher, Walid
    Qiu, Weimin
    Kassem, Moustapha
    BONE, 2015, 70 : 28 - 36
  • [23] Mitochondrial Respiration is Required for the Adipogenic Differentiation of Human Mesenchymal Stem Cells
    Zhang, Yanmin
    Toth, Peter T.
    Marsboom, Glenn
    Paul, Jonathan D.
    Rehman, Jalees
    FASEB JOURNAL, 2011, 25
  • [24] ERK signaling pathways regulate the osteogenic differentiation of human mesenchymal stem cells on collagen I and vitronectin
    Salasznyk, RM
    Klees, RF
    Hughlock, MK
    Plopper, GE
    CELL COMMUNICATION AND ADHESION, 2004, 11 (5-6): : 137 - 153
  • [25] MicroRNAs regulate signaling pathways in osteogenic differentiation of mesenchymal stem cells (Review)
    Peng, Shuping
    Gao, Dan
    Gao, Chengde
    Wei, Pingpin
    Niu, Man
    Shuai, Cijun
    MOLECULAR MEDICINE REPORTS, 2016, 14 (01) : 623 - 629
  • [26] HIV protease inhibitors selectively inhibit osteoblast differentiation and induce premature senescence in human bone marrow mesenchymal stem cells
    Lagathu, C.
    Caron-Debarle, M.
    Capeau, J.
    ANTIVIRAL THERAPY, 2010, 15 (08) : A11 - A12
  • [27] HIV protease inhibitors selectively inhibit osteoblast differentiation and induce premature senescence in human bone marrow mesenchymal stem cells
    Lagathu, C.
    Caron-Debarle, M.
    Capeau, J.
    ANTIVIRAL THERAPY, 2010, 15 : A11 - A12
  • [28] FGF23 Counters Osteoblast Differentiation in Human Mesenchymal Stem Cells by Inhibiting Vitamin D Signaling and Metabolism
    Bertucci, Christopher
    Meng, Fangang
    Zhou, Shuanhu
    Glowacki, Julie
    JOURNAL OF BONE AND MINERAL RESEARCH, 2018, 33 : 250 - 250
  • [29] Effects of melatonin on adult human mesenchymal stem cells in osteoblastic differentiation. An experimental in vitro study
    Calvo-Guirado, J. L.
    Perez-Albacete, C.
    Perez Sanchez, C.
    Boquete-Castro, A.
    Mate-Sanchez de Val, J. E.
    Delgado Pena, J. E.
    Ramirez Fernandez, M. P.
    Garces, M.
    Meseguer-Olmo, L.
    Gomez Moreno, G.
    JOURNAL OF OSSEOINTEGRATION, 2015, 7 (02) : 23 - 32
  • [30] Proteomic identification of differently expressed proteins responsible for osteoblast differentiation from human mesenchymal stem cells
    Ai-Xia Zhang
    Wei-Hua Yu
    Bao-Feng Ma
    Xin-Bing Yu
    Frank Fuxiang Mao
    Wei Liu
    Jia-Qing Zhang
    Xiu-Ming Zhang
    Shu-Nong Li
    Ming-Tao Li
    Bruce T. Lahn
    Andy Peng Xiang
    Molecular and Cellular Biochemistry, 2007, 304 : 167 - 179