ABSTRACT
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.
Subject(s)
Cell Differentiation/drug effects , NFATC Transcription Factors/metabolism , Osteoclasts/drug effects , Phenanthrenes/pharmacology , Proto-Oncogene Proteins c-fos/metabolism , Abietanes , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Bone Marrow Cells/metabolism , Carrier Proteins/genetics , Carrier Proteins/metabolism , Carrier Proteins/pharmacology , Cells, Cultured , Down-Regulation/drug effects , Gene Expression/drug effects , Gene Expression/genetics , Immunoblotting , Macrophage Colony-Stimulating Factor/pharmacology , Male , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Membrane Glycoproteins/pharmacology , Mice , Mice, Inbred ICR , NFATC Transcription Factors/genetics , Osteoclasts/cytology , Osteoclasts/metabolism , Proto-Oncogene Proteins c-fos/genetics , RANK Ligand , Receptor Activator of Nuclear Factor-kappa B , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Long term and repeated exposure of ultraviolet light on the skin often induces chronic skin diseases such as skin cancer as well as photoaging, and the mechanisms of these skin damages are closely associated with up-regulation of matrix metalloproteinase's (MMPs) activities. The methylene chloride soluble fraction of methanol extract from the stems of Styrax japonica. (Styracaceae) showed significant MMP-1 inhibition in primary human skin fibroblasts cause by ultraviolet irradiation. Four triterpenoids were isolated by column chromatography. Among them, Erythrodiol-3-acetate reduced of MMP-1 and induced of type 1 procollagen at the protein levels in a dose-dependent manner.
Subject(s)
Collagen Type I/metabolism , Matrix Metalloproteinase Inhibitors , Protease Inhibitors/pharmacology , Skin/drug effects , Styrax/chemistry , Triterpenes/pharmacology , Ultraviolet Rays , 3T3 Cells , Animals , Blotting, Western , Cells, Cultured , Fibroblasts/cytology , Fibroblasts/drug effects , Fibroblasts/enzymology , Fibroblasts/metabolism , Fibroblasts/radiation effects , Humans , Luciferases/metabolism , Mice , Protease Inhibitors/isolation & purification , Skin/cytology , Skin/enzymology , Skin/metabolism , Skin/radiation effects , Triterpenes/isolation & purificationABSTRACT
Osteoclasts, multinuclear cells specialized for bone resorption, differentiate from the monocyte/macrophage lineage of hematopoietic cells. Intervention in osteoclast differentiation is considered an effective therapeutic approach to the treatment of bone diseases involving osteoclasts. In this study, we found that tanshinone IIA, originating from Salvia miltiorrhiza Bunge, inhibited the differentiation of osteoclasts. Addition of tanshinone IIA to the osteoclast precursor culture caused a significant decrease in the level of calcitonin receptor, c-Src, and integrin beta3 mRNA, which are normally upregulated during the osteoclast differentiation dependent on RANKL (receptor activator of nuclear factor kappa B ligand). RANKL activated the ERK, Akt, and NF-kappaB signal transduction pathways in osteoclast precursor cells, and tanshinone IIA suppressed this activation. Tanshinone IIA also inhibited the bone resorptive activity of differentiated osteoclasts, which was accompanied with the disruption of the actin ring. Thus, tanshinone IIA has the potential to ameliorate bone-resorption diseases in vivo by reducing both the number and activity of osteoclasts.
