Abstract
Amylin(1–8), a cyclic peptide consisting of the eight N-terminal amino acids of the 37-amino acid peptide amylin, has been shown to induce proliferation of primary osteoblasts and to induce bone formation in healthy male mice, whereas no data on efficacy in bone disease-related models have been reported. Therefore, we evaluated any effects of amylin(1–8) in ovariectomized rats with established osteopenia, a model for postmenopausal osteoporosis. At doses up to 100 nmol/kg/day, a dose highly effective in healthy mice, amylin(1–8) was unable to increase bone mineral density in ovariectomized rats during an 8-week treatment period. Histomorphometric analysis of the tibia indicated that amylin(1–8) did not change bone histomorphometric parameters. In an attempt to verify any potential biological effects of amylin(1–8), we investigated the efficacy of this peptide in various in vitro assays. Experiments designed to confirm previously published results on the proliferative effects of amylin(1–8) on primary osteoblasts failed to show any response. Amylin(1–8) was able to partially displace 125I-rat amylin(1–37) from amylin receptors composed of the calcitonin receptor and RAMP1, indicating specific interaction of the peptide with the amylin binding site. However, in vitro efficacy assays with amylin(1–8) in calcitonin receptor-RAMP-positive HEK293T and MCF7 cells failed to reveal any agonist activity of amylin(1–8), whereas amylin(1–37) showed the expected agonist activity. In conclusion, our results indicate that amylin(1–8) does not show agonist activity on amylin receptors, does not affect osteoblast proliferation, and is devoid of anabolic activity in bone.
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Acknowledgments
We thank Karin Larsen, Christina Hansen, Claes Ekström, Jesper Rasmussen, Kennet Neist Christjansen, and Anne Qvist Rasmussen for expert technical assistance; Sritharan Shanmugam and Kirsten Ebbehøj for help with the measurements of dynamic light scattering; and Morten Karsdal for the measurement of cartilage markers.
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M. T. Ramirez, M. Stahlhut, J. S. Petersen, R. Just, and C. Thorkildsen are or were employees of Zealand Pharma at the time this study was performed. J. S. Petersen has stock ownership in Zealand Pharma. M. Ellegaard received salary from Zealand Pharma as a temporary employee. N. R. Jørgensen, P. Schwarz, and S. Petersen have stated that they have no conflict of interest.
An erratum to this article can be found at http://dx.doi.org/10.1007/s00223-010-9342-7
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Ellegaard, M., Thorkildsen, C., Petersen, S. et al. Amylin(1–8) is Devoid of Anabolic Activity in Bone. Calcif Tissue Int 86, 249–260 (2010). https://doi.org/10.1007/s00223-010-9338-3
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DOI: https://doi.org/10.1007/s00223-010-9338-3