title: Regulation of matrix metalloproteinase-9 protein expression by 1α, 25-(OH)₂D₃ during osteoclast differentiation.
abstract: To investigate 1α,25-(OH)₂D₃ regulation of matrix metalloproteinase-9 (MMP-9) protein expression during osteoclast formation and differentiation, receptor activator of nuclear factor kB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) were administered to induce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of 1α,25-(OH)₂D₃ during culturing, and cell proliferation was measured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistant acid phosphatase (TRAP) staining and assessing bone lacunar resorption. MMP-9 protein expression levels were measured with Western blotting. We showed that 1α,25-(OH)₂D₃ inhibited RAW264.7 cell proliferation induced by RANKL and M-CSF, increased the numbers of TRAP-positive osteoclasts and their nuclei, enhanced osteoclast bone resorption, and promoted MMP-9 protein expression in a concentration-dependent manner. These findings indicate that 1α,25-(OH)₂D₃ administered at a physiological relevant concentration promoted osteoclast formation and could regulate osteoclast bone metabolism by increasing MMP-9 protein expression during osteoclast differentiation.
authors.affiliation: College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
authors.forename: Jian-Hong
authors.initials: JH
authors.lastname: Gu
authors.forename: Xi-Shuai
authors.initials: XS
authors.lastname: Tong
authors.forename: Guo-Hong
authors.initials: GH
authors.lastname: Chen
authors.forename: Xue-Zhong
authors.initials: XZ
authors.lastname: Liu
authors.forename: Jian-Chun
authors.initials: JC
authors.lastname: Bian
authors.forename: Yan
authors.initials: Y
authors.lastname: Yuan
authors.forename: Zong-Ping
authors.initials: ZP
authors.lastname: Liu
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002117:Calcitriol / Q000494:pharmacology*; D002120:Calcium Channel Agonists / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D007527:Isoenzymes / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D010010:Osteoclasts / Q000166:cytology* / Q000201:enzymology*; D000071681:Tartrate-Resistant Acid Phosphatase; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D002120:Calcium Channel Agonists; D007527:Isoenzymes; D013778:Tetrazolium Salts; D013844:Thiazoles; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase; D020780:Matrix Metalloproteinase 9; C022616:thiazolyl blue; D002117:Calcitriol
keywords: 1α,25-(OH)₂D₃; MMP-9; TRAP; bone lacunar resorption; osteoclast