Effect of beta-alanyl-L-histidinato zinc on the differentiation of C2C12 cells.
Although beta-alanyl-L-histidinato zinc (AHZ) can promote osteoblast differentiation, the molecular mechanism responsible is not fully understood. The purpose of this study was to determine the effect of AHZ on undifferentiating mesenchymal cells. C2C12, a typical pluripotential mesenchymal cell line, was used. The cells were cultured in 5% serum-containing medium to induce differentiation, either with or without the addition of AHZ. Cell lineage was determined by immunostaining of type II myosin heavy chains, alkaline phosphatase (ALPase) activity, mRNA expression of cellular phenotype-specific markers using semi-quantitative reverse transcriptase-polymerase chain reaction, and core binding factor alpha1/runt-related transcription factor-2 (Cbfa1/Runx2) protein synthesis using Western blot analysis. C2C12 cells cultured in the presence of AHZ were strongly inhibited from developing into myoblasts, and showed high ALPase activity that was approximately double that in the vehicle. The expression of mRNA for Cbfa1/Runx2, ALPase, Sox9 and type X collagen was increased markedly by the AHZ-stimulated medium, whereas that of desmin and MyoD mRNA was drastically decreased. AHZ increased Cbfa1/Runx2 protein expression substantially. These results provide clear evidence that AHZ converts the differentiation pathway of C2C12 cells to the osteoblast and/or chondroblast lineage.
Takada
Tsuyoshi
T
Department of Biochemistry, Nihon University School of Dentistry, Tokyo 101-8310, Japan.
Suzuki
Naoto
N
Ito-Kato
Emi
E
Noguchi
Yoshihiro
Y
Ito
Mitsuyasu
M
Maeno
Masao
M
Otsuka
Kichibee
K
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D015153:Blotting, Western; D002336:Carnosine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D024121:Collagen Type X / Q000096:biosynthesis; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009124:Muscle Proteins / Q000096:biosynthesis; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009687:Nuclear Proteins / Q000096:biosynthesis; D009942:Organometallic Compounds / Q000494:pharmacology*; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D014157:Transcription Factors / Q000096:biosynthesis; D017967:Zinc Compounds
D024121:Collagen Type X; D003470:Culture Media; D009124:Muscle Proteins; D009687:Nuclear Proteins; D009942:Organometallic Compounds; D012333:RNA, Messenger; D014157:Transcription Factors; D017967:Zinc Compounds; C061957:polaprezinc; D002336:Carnosine; D000469:Alkaline Phosphatase