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B Ligand, Osteoprotegerin, and Receptor Activator of NF-B in Mouse Calvariae1
* Department of Oral Cell Biology, Umea University, Umea, Sweden;
Center for Musculoskeletal Research, National Institute for Working Life, Umea, Sweden;
Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR 72205
IL-6, leukemia inhibitory factor (LIF), and oncostatin M (OSM) are
IL-6-type cytokines that stimulate osteoclast formation and function.
In the present study, the resorptive effects of these agents and their
regulation of receptor activator of NF-
B ligand (RANKL), RANK, and
osteoprotegerin (OPG) were studied in neonatal mouse calvaria. When
tested separately, neither human (h) IL-6 nor the human soluble IL-6R
(shIL-6R) stimulated bone resorption, but when hIL-6 and the shIL-6R
were combined, significant stimulation of both mineral and matrix
release from bone explants was noted. Semiquantitative RT-PCR showed
that hIL-6 plus shIL-6R enhanced the expression of RANKL and OPG in
calvarial bones, but decreased RANK expression. Human LIF, hOSM, and
mouse OSM (mOSM) also stimulated 45Ca release and enhanced
the mRNA expression of RANKL and OPG in mouse calvaria, but had no
effect on the expression of RANK. In agreement with the RT-PCR
analyses, ELISA measurements showed that both hIL-6 plus shIL-6R and
mOSM increased RANKL and OPG proteins. 1,25-Dihydroxyvitamin
D3 (D3) also increased the RANKL protein level,
but decreased the protein level of OPG. OPG inhibited 45Ca
release stimulated by RANKL, hIL-6 plus shIL-6R, hLIF, hOSM, mOSM, and
D3. An Ab neutralizing mouse gp130 inhibited
45Ca release induced by hIL-6 plus shIL-6R. These
experiments demonstrated stimulation of calvarial bone resorption and
regulation of mRNA and protein expression of RANKL and OPG by
D3 and IL-6 family cytokines as well as regulation of RANK
expression in preosteoclasts/osteoclasts of mouse calvaria by
D3 and hIL-6 plus shIL-6R.
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