Abstract
Exendin-4 was found to be beneficial to the skeleton in diabetic rodents. In this study, we assessed the changes of bone mineral densities (BMDs) and quality in non-diabetic ovariectomized (OVX) rats after treatment with exendin-4. The regulatory role of exendin-4 on osteoblastogenesis and adipogenesis in rat bone marrow stromal cells (BMSCs) was also explored. Three months after sham surgery or OVX, 18 5-month-old female Wistar rats were divided into three groups and received the following treatment for 8 weeks: (1) Sham + vehicle; (2) OVX + vehicle; and (3) OVX + exendin-4 20 µg/kg/day. Micro-CT and three-point bending test were used to evaluate the BMDs, bone morphometric parameters, and biomechanical properties. Real-time PCR and Western blot were performed to measure gene and protein expression after exendin-4 treatment in adipogenesis and osteoblastogenesis of rat BMSCs. Exendin-4 could improve trabecular volume, thickness, and number, increase BMD, and reduce trabecular spacing in the lumbar spine and femur of OVX rats. Exendin-4 had little impact on the mechanical resistance of femurs to fracture. When rat BMSCs were treated with exendin-4, the mRNA expression levels of runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and collagen α1 (Coll-1) were increased, while those of peroxisome proliferators activated receptor γ (PPARγ) and CCAAT/enhancer-binding protein (C/EBPα) decreased. Exendin-4 treatment also resulted in increased expression levels of p38, p42/44, and β-catenin proteins. Exendin-4 was anabolic to bone in OVX rats possibly by facilitating osteoblastogenesis while repressing adipogenesis during BMSC lineage differentiation.
Similar content being viewed by others
References
S. Chon, J.P. Riveline, B. Blondeau, J.F. Gautier, Incretin-based therapy and pancreatic beta cells. Diabetes Metab 40(6), 411–422 (2014). doi:10.1016/j.diabet.2014.05.003
K. Mather, Extrapancreatic effects of GLP-1 and other incretins. Revendocrine Metab. Disord. 15(3), 169 (2014). doi:10.1007/s11154-014-9292-x
B. Nuche-Berenguer, S. Portal-Nunez, P. Moreno, N. Gonzalez, A. Acitores, A. Lopez-Herradon, P. Esbrit, I. Valverde, M.L. Villanueva-Penacarrillo, Presence of a functional receptor for GLP-1 in osteoblastic cells, independent of the cAMP-linked GLP-1 receptor. J. Cell. Physiol. 225(2), 585–592 (2010). doi:10.1002/jcp.22243
C. Sanz, P. Vazquez, C. Blazquez, P.A. Barrio, M.D.M. Alvarez, E. Blazquez, Signaling and biological effects of glucagon-like peptide 1 on the differentiation of mesenchymal stem cells from human bone marrow. Am J Physiol. Endocrinol Metab 298(3), E634–643 (2010). doi:10.1152/ajpendo.00460.2009
C. Brand, D. Abdel-Atti, Y. Zhang, S. Carlin, S.M. Clardy, E.J. Keliher, W.A. Weber, J.S. Lewis, T. Reiner, In vivo imaging of GLP-1R with a targeted bimodal PET/fluorescence imaging agent. Bioconjug. Chem. 25(7), 1323–1330 (2014). doi:10.1021/bc500178d
B. Nuche-Berenguer, P. Moreno, P. Esbrit, S. Dapia, J.R. Caeiro, J. Cancelas, J.J. Haro-Mora, M.L. Villanueva-Penacarrillo, Effect of GLP-1 treatment on bone turnover in normal, type 2 diabetic, and insulin-resistant states. Calcif. Tissue Int. 84(6), 453–461 (2009). doi:10.1007/s00223-009-9220-3
A. Papazafiropoulou, N. Papanas, S. Pappas, E. Maltezos, Role of endogenous GLP-1 and its agonists in osteopenia and osteoporosis: but we little know until tried. Curr Diabetes Rev. 10(1), 43–47 (2014)
B. Su, H. Sheng, M. Zhang, L. Bu, P. Yang, L. Li, F. Li, C. Sheng, Y. Han, S. Qu, J. Wang, Risk of bone fractures associated with glucagon-like peptide-1 receptor agonists’ treatment: a meta-analysis of randomized controlled trials. Endocrine (2014). doi:10.1007/s12020-014-0361-4
B. Nuche-Berenguer, P. Moreno, S. Portal-Nunez, S. Dapia, P. Esbrit, M.L. Villanueva-Penacarrillo, Exendin-4 exerts osteogenic actions in insulin-resistant and type 2 diabetic states. Regul. Pept. 159(1–3), 61–66 (2010). doi:10.1016/j.regpep.2009.06.010
B. Nuche-Berenguer, D. Lozano, I. Gutierrez-Rojas, P. Moreno, M.L. Marinoso, P. Esbrit, M.L. Villanueva-Penacarrillo, GLP-1 and exendin-4 can reverse hyperlipidic-related osteopenia. J Endocrinol 209(2), 203–210 (2011). doi:10.1530/JOE-11-0015
X. Ma, J. Meng, M. Jia, L. Bi, Y. Zhou, Y. Wang, J. Hu, G. He, X. Luo, Exendin-4, a glucagon-like peptide-1 receptor agonist, prevents osteopenia by promoting bone formation and suppressing bone resorption in aged ovariectomized rats. J Bone Miner Res 28(7), 1641–1652 (2013). doi:10.1002/jbmr.1898
H. Taipaleenmaki, B.M. Abdallah, A. AlDahmash, A.M. Saamanen, M. Kassem, Wnt signalling mediates the cross-talk between bone marrow derived pre-adipocytic and pre-osteoblastic cell populations. Exp. Cell Res. 317(6), 745–756 (2011). doi:10.1016/j.yexcr.2010.12.015
B.M. Abdallah, M. Kassem, New factors controlling the balance between osteoblastogenesis and adipogenesis. Bone 50(2), 540–545 (2012). doi:10.1016/j.bone.2011.06.030
K. Tatarkiewicz, P. Belanger, G. Gu, D. Parkes, D. Roy, No evidence of drug-induced pancreatitis in rats treated with exenatide for 13 weeks. Diabetes Obes. Metab. 15(5), 417–426 (2013). doi:10.1111/dom.12040
D. Roy, K.D. Chadwick, K. Tatarkiewicz, C. LaCerte, A.M. Bergholm, T. Brodie, R.S. Mangipudy, D. Parkes, M.J. Graziano, T.P. Reilly, The glucagon-like peptide-1-based therapeutics exenatide and saxagliptin did not cause detrimental effects on the pancreas in mice, rats, dogs and monkeys. Diabetes Obes. Metab. 16(10), 910–921 (2014). doi:10.1111/dom.12294
C.P. Liang, S. Han, G. Li, I. Tabas, A.R. Tall, Impaired MEK signaling and SERCA expression promote ER stress and apoptosis in insulin-resistant macrophages and are reversed by exenatide treatment. Diabetes 61(10), 2609–2620 (2012). doi:10.2337/db11-1415
J. Taher, C.L. Baker, C. Cuizon, H. Masoudpour, R. Zhang, S. Farr, M. Naples, C. Bourdon, Z. Pausova, K. Adeli, GLP-1 receptor agonism ameliorates hepatic VLDL overproduction and de novo lipogenesis in insulin resistance. Mol Metab 3(9), 823–833 (2014). doi:10.1016/j.molmet.2014.09.005
Q. Zhao, X. Shen, W. Zhang, G. Zhu, J. Qi, L. Deng, Mice with increased angiogenesis and osteogenesis due to conditional activation of HIF pathway in osteoblasts are protected from ovariectomy induced bone loss. Bone 50(3), 763–770 (2012). doi:10.1016/j.bone.2011.12.003
N. Jaiswal, S.E. Haynesworth, A.I. Caplan, S.P. Bruder, Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J. Cell. Biochem. 64(2), 295–312 (1997)
X. Ma, L. Xu, S. Wang, B. Cui, X. Li, J. Xu, G. Ning, Deletion of steroid receptor coactivator-3 gene ameliorates hepatic steatosis. J. Hepatol. 55(2), 445–452 (2011). doi:10.1016/j.jhep.2010.11.022
I. Dicembrini, E. Mannucci, C.M. Rotella, Bone: incretin hormones perceiver or receiver? Exp Diabetes Res 2012, 519784 (2012). doi:10.1155/2012/519784
C. Yamada, Y. Yamada, K. Tsukiyama, K. Yamada, N. Udagawa, N. Takahashi, K. Tanaka, D.J. Drucker, Y. Seino, N. Inagaki, The murine glucagon-like peptide-1 receptor is essential for control of bone resorption. Endocrinology 149(2), 574–579 (2008). doi:10.1210/en.2007-1292
G. Mabilleau, A. Mieczkowska, N. Irwin, P.R. Flatt, D. Chappard, Optimal bone mechanical and material properties require a functional glucagon-like peptide-1 receptor. J Endocrinol 219(1), 59–68 (2013). doi:10.1530/JOE-13-0146
J.Y. Kim, S.K. Lee, K.J. Jo, D.Y. Song, D.M. Lim, K.Y. Park, L.F. Bonewald, B.J. Kim, Exendin-4 increases bone mineral density in type 2 diabetic OLETF rats potentially through the down-regulation of SOST/sclerostin in osteocytes. Life Sci. 92(10), 533–540 (2013). doi:10.1016/j.lfs.2013.01.001
E.L. Pacheco-Pantoja, L.R. Ranganath, J.A. Gallagher, P.J. Wilson, W.D. Fraser, Receptors and effects of gut hormones in three osteoblastic cell lines. BMC Physiol 11, 12 (2011). doi:10.1186/1472-6793-11-12
T. Akune, S. Ohba, S. Kamekura, M. Yamaguchi, U.I. Chung, N. Kubota, Y. Terauchi, Y. Harada, Y. Azuma, K. Nakamura, T. Kadowaki, H. Kawaguchi, PPARgamma insufficiency enhances osteogenesis through osteoblast formation from bone marrow progenitors. J. Clin. Investig. 113(6), 846–855 (2004). doi:10.1172/JCI19900
H.J. Wee, G. Huang, K. Shigesada, Y. Ito, Serine phosphorylation of RUNX2 with novel potential functions as negative regulatory mechanisms. EMBO Rep. 3(10), 967–974 (2002). doi:10.1093/embo-reports/kvf193
G. Xiao, D. Jiang, R. Gopalakrishnan, R.T. Franceschi, Fibroblast growth factor 2 induction of the osteocalcin gene requires MAPK activity and phosphorylation of the osteoblast transcription factor, Cbfa1/Runx2. J. Biol. Chem. 277(39), 36181–36187 (2002). doi:10.1074/jbc.M206057200
T.J. Rosol, On-target effects of GLP-1 receptor agonists on thyroid C-cells in rats and mice. Toxicol. Pathol. 41(2), 303–309 (2013). doi:10.1177/0192623312472402
F. Boess, C. Bertinetti-Lapatki, S. Zoffmann, C. George, T. Pfister, A. Roth, S.M. Lee, W.E. Thasler, T. Singer, L. Suter, Effect of GLP1R agonists taspoglutide and liraglutide on primary thyroid C-cells from rodent and man. J. Mol. Endocrinol. 50(3), 325–336 (2013). doi:10.1530/JME-12-0186
Acknowledgments
This study was supported by the National Nature Science Foundation of China (No. 81370977, 81170804, 81370018) and the Shanghai Municipal Health Bureau Project (2012-235).
Conflict of interest
All authors state that they have no conflicts of interest.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Han-xiao Sun, and Nan Lu have contributed equally to this work.
Rights and permissions
About this article
Cite this article
Sun, Hx., Lu, N., Liu, Dm. et al. The bone-preserving effects of exendin-4 in ovariectomized rats. Endocrine 51, 323–332 (2016). https://doi.org/10.1007/s12020-015-0667-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12020-015-0667-x