title: Effects of 1α,25-(OH)2D3 on the formation and activity of osteoclasts in RAW264.7 cells.
abstract: The hormonally active form of vitamin D3, 1α,25-(OH)2D3, has an important role in bone metabolism. This study examined the effects of 1α,25-(OH)2D3 on the ability of two cytokines, receptor activator of nuclear factor-κB ligand (RANKL) and macrophage-colony stimulating factor (M-CSF), to induce RAW 264.7 cells to form osteoclasts. A TRAP histochemical staining assay and bone resorption analysis were used to identify the rate of formation and activity of osteoclasts. The numbers of osteoclasts formed, and their bone resorption activity, was enhanced by the addition of 1α,25-(OH)2D3. The expression levels of osteoclast-specific proteins that are essential for bone resorption, integrin β3, V-ATPase, CAII, CTSK, TRAP and MMP-9, were detected by western blotting. During 48 h, the expression levels of all these proteins significantly increased. Quantitative real-time polymerase chain reaction was used to determine the expression levels of the transcription factors, c-Fos and NFATcl. The expression levels of c-Fos and NFATc1 also increased 24h after treatment with 1α,25-(OH)2D3. These results suggest that 1α,25-(OH)2D3 can regulate bone metabolism by directly enhancing the formation and maturation of osteoclasts.
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
authors: Jianhong
authors: J
authors: Gu
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
authors: Xi-Shuai
authors: XS
authors: Tong
authors: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
authors: Guo-Hong
authors: GH
authors: Chen
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
authors: Dong
authors: D
authors: Wang
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
authors: Yang
authors: Y
authors: Chen
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
authors: Yan
authors: Y
authors: Yuan
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
authors: Xue-Zhong
authors: XZ
authors: Liu
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. Electronic address: bianjianchun@sina.com.
authors: Jian-Chun
authors: JC
authors: Bian
authors: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
authors: Zong-Ping
authors: ZP
authors: Liu
mesh_terms: D000818:Animals; D001862:Bone Resorption / Q000378:metabolism*; D001842:Bone and Bones / Q000378:metabolism; D056657:Cathepsin K / Q000096:biosynthesis; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002762:Cholecalciferol / Q000031:analogs & derivatives / Q000494:pharmacology*; D039661:Integrin beta3 / Q000096:biosynthesis; D016173:Macrophage Colony-Stimulating Factor / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis; D056892:Mediator Complex / Q000096:biosynthesis; D051379:Mice; D050778:NFATC Transcription Factors / Q000096:biosynthesis; D010010:Osteoclasts / Q000166:cytology*; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis; D053245:RANK Ligand / Q000378:metabolism*; D025262:Vacuolar Proton-Translocating ATPases / Q000096:biosynthesis
chemical_list: D039661:Integrin beta3; D056892:Mediator Complex; D050778:NFATC Transcription Factors; C494005:Nfatc1 protein, mouse; D016760:Proto-Oncogene Proteins c-fos; D053245:RANK Ligand; C505689:Tnfsf11 protein, mouse; D002762:Cholecalciferol; D016173:Macrophage Colony-Stimulating Factor; D056657:Cathepsin K; C534132:Ctsk protein, mouse; D020780:Matrix Metalloproteinase 9; D025262:Vacuolar Proton-Translocating ATPases
keywords: 1α,; 25-(OH)(2)D(3); Bone resorption; OPG; Osteoclasts; RANKL; RAW264.7 cells