Fluid flow induces Rankl expression in primary murine calvarial osteoblasts

Journal of Cellular Biochemistry
Meenal MehrotraCarol C Pilbeam

Abstract

Mechanical loading of bone generates fluid flow within the mineralized matrix that exerts fluid shear stress (FSS) on cells. We examined effects of FSS on receptor activator of nuclear factor kappa B ligand (RANKL), a critical factor for osteoclast formation. Primary murine osteoblasts were subjected to pulsatile FSS (5 Hz, 10 dynes/cm(2)) for 1 h and then returned to static culture for varying times (post-FSS). Protein levels were measured by Western analysis and mRNA by Northern analysis, RT-PCR and quantitative PCR. There were 20- to 40-fold increases in RANKL mRNA at 2-4 h post-FSS. RANKL protein was induced by 2 h post-FSS and remained elevated for at least 8 h. Effects were independent of cyclooxygenase-2 activity. Small increases (up to three-fold) in mRNA of the decoy receptor for RANKL, osteoprotegerin, were seen. Five min of FSS, followed by static culture, was as effective in stimulating RANKL mRNA as 4 h of continuous FSS. FSS induced cAMP activity, and H-89, a protein kinase A (PKA) inhibitor, blocked the FSS induction of RANKL. H-89 also inhibited the PKC pathway, but specific PKC inhibitors, GF109203X and Go6983, did not inhibit FSS-induced RANKL. FSS induced phosphorylation of ERK1/2, and PD98059, an inhibitor o...Continue Reading

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Citations

Aug 9, 2011·Journal of Bone and Mineral Metabolism·Ingrid BergströmJonas Brinck
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Sep 6, 2012·Journal of Cellular Biochemistry·Zhang Qing HongLiu Li
Jan 8, 2013·International Journal of Molecular Medicine·Sumika KameyamaJunichiro Iida
Oct 26, 2016·Molecular Medicine Reports·Masaaki IkedaJunichiro Iida

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