Isolation of GnafC, a polysaccharide constituent of Gnaphalium affine, and synergistic effects of GnafC and ascorbate on the phenotypic expression of osteoblastic MC3T3-E1 cells.
After screening extensively factors in plant extracts that increase alkaline phosphatase activity, an osteoblastic differentiation marker protein in mouse calvarial osteoblast MC3T3-E1 cells, GnafC derived from Gnaphalium affine, was found to significantly enhance the alkaline phosphatase (ALPase) activity in a synergistic manner with ascorbate. GnafC was a polysaccharaide with an approximate molecular mass of 10,000 and comprised mannose, xylose, arabinose, galactose and glucose in a molar ratio of 1:2:4.3:2.5:2.7. Expression of the osteoblastic differentiation marker genes was examined by semiquantitative RT-PCR with RNAs prepared from cells at different developmental stages. With ascorbate in the culture, GnafC enhanced the expression of the ALPase and MMP13 genes from the early stage of differentiation, leading to maturation of the collagenous extracellular matrix (ECM), a prerequisite for mineralization.
Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan.
Yukihiro
Y
Aoshima
Yoshinori
Y
Hasegawa
Sho
S
Hasegawa
Atsushi
A
Nagasaka
Tomofumi
T
Kimura
Satofumi
S
Hashimoto
Yasuyoshi
Y
Torii
Norihiro
N
Tsukagoshi
D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D017364:Collagenases / Q000096:biosynthesis / Q000235:genetics; D004357:Drug Synergism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D053509:Matrix Metalloproteinase 13; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects*; D010641:Phenotype; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology; D011134:Polysaccharides / Q000302:isolation & purification* / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis
C487922:GnafC saccharide; D010936:Plant Extracts; D011134:Polysaccharides; D012333:RNA, Messenger; D000469:Alkaline Phosphatase; D017364:Collagenases; D053509:Matrix Metalloproteinase 13; C507081:Mmp13 protein, mouse; D001205:Ascorbic Acid