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Am J Physiol Regul Integr Comp Physiol 292: R2208-R2215, 2007. First published March 22, 2007; doi:10.1152/ajpregu.00013.2007
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DEVELOPMENTAL PHYSIOLOGY AND PREGNANCY

Melatonin inhibits fatty acid-induced triglyceride accumulation in ROS17/2.8 cells: implications for osteoblast differentiation and osteoporosis

M. Sanchez-Hidalgo,1,* Z. Lu,2,* D.-X. Tan,1 M. D. Maldonado,1 R. J. Reiter,1 and R. I. Gregerman2

1Department of Cellular and Structural Biology, University of Texas Health Science Center at San Antonio, Texas; and 2Geriatric Research Education and Clinical Center, South Texas Veterans Health Care System, Audie L. Murphy Division, and Division of Geriatrics and Gerontology, Department of Medicine, University of Texas Health Science Center at San Antonio, Texas

Submitted 9 January 2007 ; accepted in final form 6 March 2007

Melatonin is produced not only by the pineal gland but by cells of the bone marrow. Moreover, melatonin is known to promote osteogenic differentiation in several cell line models and in multipotential bone marrow mesenchymal stem cells. Fatty acids have been independently shown to direct such cells to acquire the phenotype and molecular characteristics of adipocytes. To examine the effect of melatonin on intracellular triglyceride accumulation, an indicator of adipogenic differentiation in the rat osteoblast-like ROS17/2.8 cell line, cells were incubated with added oleic acid (100 µM), fixed and stained with Oil Red O. Cellular lipid accumulation was quantitated by an Oil Red O method highly specific for triglycerides and expressed as a triglyceride accumulation index (TGAI, triglyceride per cell). Melatonin in nanomolar concentrations inhibited oleic acid-induced triglyceride accumulation. To identify the mechanism by which melatonin reduces triglyceride accumulation, cells were incubated with the two melatonin receptor antagonists, luzindole and S20928, or forskolin, a stimulator of adenylyl cyclase and cAMP production. These compounds prevented the inhibitory effect of melatonin on triglyceride accumulation, indicating that melatonin acts through known melatonin receptor-mediated mechanisms. In view of the previously demonstrated positive effects of melatonin in promoting osteoblastic differentiation in ROS17/2.8 cells and their reciprocal adipocytic differentiation induced by fatty acids, our observations may serve to relate the known age-related decreases of melatonin production, the shift in the bone marrow toward an adipocytic line of cell development, and the development of osteoporosis during aging.

fatty acids; oleic acid; osteogenesis; luzindole; S20928



Address for reprint requests and other correspondence: R. I. Gregerman, Div. of Geriatrics and Gerontology, Dept. of Medicine, Univ. of Texas Health Science Center at San Antonio, TX 78229-3900 (e-mail: gregerman{at}uthscsa.edu) or R. J. Reiter, Dept. of Cellular and Structure Biology, Univ. of Texas Health Science Center at San Antonio, TX 78229-3900 (e-mail: reiter{at}uthscsa.edu)







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