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Concentrations of Calcium-binding Protein and Bone Gla-protein in Culture Medium and CaBP mRNA Expression in Osteoblasts of Broiler Chickens
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 Title & Authors
Concentrations of Calcium-binding Protein and Bone Gla-protein in Culture Medium and CaBP mRNA Expression in Osteoblasts of Broiler Chickens
Guo, Xiaoyu; Yan, Sumei; Shi, Binlin; Feng, Yongmiao;
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This study was conducted to determine the effects of excess vitamin A on alkaline phosphatase (ALP) activity, contents of calcium-binding protein (CaBP), bone gla-protein (BGP) in culture medium and CaBP mRNA expression in chicken osteoblasts in vitro. Osteoblastic cells in the tibia from 1-day-old Arbor Acre broiler chickens were isolated using enzyme digestion. The subconfluenced cells were divided into eight treatments with six replicates in each treatment and cultured in a medium containing either vehicle or different levels of vitamin A (0, 0.2, 0.6, 1.0, 2.0, 5.0, 10.0 and /ml), and the control received an equivalent volume of ethanol. The incubation lasted 48 h. The results showed that vitamin A down-regulated ALP activity in the culture medium as well as CaBP mRNA expression of osteoblasts in a linear dose-dependent manner (p = 0.124 and p<0.10, respectively), and suppressed the contents of BGP and CaBP in the culture medium in a quadratic dose-dependent manner (p<0.05 and p<0.10, respectively) with increasing addition of vitamin A. The addition of 0-/ml vitamin A to the culture medium increased ALP activity, BGP and CaBP contents as well as CaBP mRNA expression compared with other groups, but positive effects of vitamin A tended to be suppressed when vitamin A was increased to /ml, and adverse effects occurred when vitamin A was increased to 10.0-/ml. These results implied that there was a threshold level of vitamin A inclusion beyond which inhibitory effects occurred, and the mechanism by which overdose of vitamin A reduced bone growth in chickens was probably reduced osteoblastic cell activity, and inhibited expression of CaBP mRNA and CaBP secretion.
Vitamin A;Broiler Chicken;Calcium-binding Protein;mRNA Expression;Osteoblast;Bone Gla-protein;
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