OBJECTIVE: Osteoclasts are bone-resorbing multinucleated cells derived from the monocyte/macrophage lineage during normal and pathological bone turnover. Recently, several studies revealed that alpha-tocopheryl succinate (alphaTP-suc) have demonstrated potent anti-cancer activities in vitro and in vivo. However, the effects of alphaTP-suc on osteoclast formation and bone resorption remain unknown. Thus, in this study, we examined the effects of alphaTP-suc on osteoclast differentiation and bone resorbing activity in inflammatory bone loss model. METHODS: Osteoclast differentiation assay was performed by cocultures of mouse bone marrow cells and calvarial osteoblasts in culture media including interleukin-1 (IL-1). Osteoclasts were stained for tartrate-resistant acid phosphatase (TRAP). The level of receptor activator of nuclear factor-kappaB ligand (RANKL) mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). ICR mice were administered an intraperitoneal injections of alphaTP-suc or dimethyl sulfoxide (DMSO) 1 day before the implantation of a freeze-dried collagen sponge loaded with phosphate-buffered saline (PBS) or IL-1 over the calvariae and every other day for 7 days. The whole calvariae were obtained and analyzed by micro-computed tomography (CT) scanning, and stained for TRAP. RESULTS: alphaTP-suc inhibits osteoclast formation in cocultures stimulated by IL-1 and decreased the level of expression of RANKL mRNA in osteoblasts. In addition, administered intraperitoneal injections of alphaTP-suc prevented IL-1-mediated osteoclast formation and bone loss in vivo. CONCLUSION: Our findings suggest that alphaTP-suc may have therapeutic value for treating and preventing bone-resorptive diseases, such as osteoporosis.
Acid Phosphatase ; Animals ; Bone Marrow Cells ; Bone Resorption ; Coculture Techniques ; Collagen ; Culture Media ; Cytokines ; Dimethyl Sulfoxide ; Injections, Intraperitoneal ; Interleukin-1 ; Isoenzymes ; Mice ; Mice, Inbred ICR ; Osteoblasts ; Osteoclasts ; Osteoporosis ; Porifera ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; RNA, Messenger ; Safrole ; Skull ; Succinic Acid

We identified a neuroprotective single fraction among 62 ones of hexane extract from Uncaria sinensis (JGH43IA) and investigated its effects and mechanisms in primary cortical neurons. Pretreatment with JGH43IA showed a significantly increase cell viability in a dose-dependent manner with a decrease in the lactate dehydrogenase release. When we performed morphological assay and flow cytometry to determination of the type of cell death, pretreatment with JGH43IA showed a significant reduction of glutamate-induced apoptotic cell death. Then we explored the downstream signaling pathways of N-methyl-D-aspartate receptor (NMDAR) with calpain activation to elucidate possible pathways of neuroprotection by JGH43IA. Pretreatment with JGH43IA exhibited a significant attenuation of NMDAR GluN2B subunit activation and a decrease in active form of calpain 1 leading to subsequent cleavage of striatal-enriched protein tyrosine phosphatase (STEP). In addition, pretreatment with JGH43IA showed a marked increase of cAMP responsive element binding protein. These results suggest that JGH43IA may have neuroprotective effects through down-regulation of NMDAR GluN2B subunit and calpain 1 activation, and subsequent alleviation of STEP cleavage. This single fraction from U. sinensis might be a useful therapeutic agent for brain disorder associated with glutamate injury.
Brain Diseases ; Calpain ; Carrier Proteins ; Cell Death ; Cell Survival ; Down-Regulation ; Flow Cytometry ; Glutamates ; Glutamic Acid ; L-Lactate Dehydrogenase ; N-Methylaspartate ; Neurons* ; Neuroprotective Agents* ; Protein Tyrosine Phosphatases ; Receptors, N-Methyl-D-Aspartate ; Uncaria*

Bone is a dynamic tissue that is regulated by the activity of bone-resorbing osteoclasts and bone-forming osteoblasts. Excessive osteoclast formation causes diseases such as osteoporosis and rheumatoid arthritis. Natural substances may be useful as therapeutic drugs to prevent many diseases in humans because they avoid the many side effects of treatment with chemical compounds. Here we show that tanshinone IIA isolated from Salvia miltiorrhiza Bunge inhibits the receptor activator of NF-kappaB ligand (RANKL)-mediated osteoclast differentiation of osteoclast precursors. Tanshinone IIA suppressed the expression levels of c-Fos and NFATc1 induced by RANKL. However, retrovirus-mediated overexpression of c-Fos induced the expression of NFATc1 despite the presence of tanshinone IIA and reversed the inhibitory effect of tanshinone IIA on osteoclast differentiation. Also, the introduction of osteoclast precursors with the NFATc1 retrovirus led to osteoclast differentiation in the presence of tanshinone IIA. Our results suggest that tanshinone IIA may have a role as a therapeutic drug in the treatment of bone disease such as osteoporosis.
Reverse Transcriptase Polymerase Chain Reaction ; Receptor Activator of Nuclear Factor-kappa B ; RANK Ligand ; Proto-Oncogene Proteins c-fos/genetics/*metabolism ; Phenanthrenes/*pharmacology ; Osteoclasts/cytology/*drug effects/metabolism ; NFATC Transcription Factors/genetics/*metabolism ; Mice, Inbred ICR ; Mice ; Membrane Glycoproteins/genetics/metabolism/pharmacology ; Male ; Macrophage Colony-Stimulating Factor/pharmacology ; Immunoblotting ; Gene Expression/drug effects/genetics ; Down-Regulation/drug effects ; Cells, Cultured ; Cell Differentiation/*drug effects ; Carrier Proteins/genetics/metabolism/pharmacology ; Bone Marrow Cells/cytology/drug effects/metabolism ; Animals