Abstract
The effect of β-alany-L-histidinato zinc (AHZ) on bone cell function was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37°C in a CO2 incubator in plastic dishes containing α-modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus AHZ (10−7–10−5 M) or other reagents, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10−7–10−5 M) produced a remarkable increase of alkaline phosphatase activity and protein concentration in osteoblastic cells. Thus increases were seen with the prolonged cultivation (12–21 days). With the culture of 1, 3 and 12 days, the effect of AHZ (10−6 M) to increase alkaline phosphatase activity and protein concentration was more intensive than the effect of zinc sulfate, (10−6 M). The AHZ effects were completely abolished by the presence of cycloheximide (10−6 M), indicating that AHZ stimulates protein synthesis in the cells. The present study suggests that AHZ has a stimulatory effect on cell differentiation, and that this effect is partly involved on protein synthesis in osteoblastic cells.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Burch RE, Hahn HKJ, Sullivan JF: Newer aspects of the roles of zinc, manganase and copper in human nutrition. Clin Chim 21: 501–520, 1975
Hurley LS, Gowan J, Milhaud G: Calcium metabolism in manganese-deficient and zinc-deficient rats. Proc Soc Exp Biol Med 130: 856–860, 1969
Oner G, Bhaumick B, Bala EM: Effect of zinc deficiency on serum somatomedin levels and skeletal growth in young rats. Endocrinology 114: 1860–1863, 1984
Yamaguchi M, Yamaguchi R: Action of zinc on bone metabolism in rats: Increases in alkaline phosphatase activity and DNA content. Biochem Pharmacol 35: 773–777, 1986
Yamaguchi M, Oishi H, Suketa Y: Sumulatory effect of zinc on bone formation in tissue culture. Biochem Pharmacol 36: 4007–4012, 1987
Yamaguchi M, Oishi H, Suketa Y: Zinc stimulation of bone protein synthesis in tissue culture: Activation of aminoacyl-tRNA synthetase. Biochem Pharmacol 37: 4075–4080, 1988
Yamaguchi M, Oishi K: A new zinc compound, β-alanyl-L-histidinato zinc, stimulates bone growth in weanling rats. Res Exp Med 190: 105–110, 1990
Yamaguchi M, Miwa H: Stimulatory effect of β-alanyl-L-histidinato zinc on bone formation in tissue culture. Pharmacology 42: 230–240, 1991
Hashizume M, Yamaguchi M: Stimulatory effect of β-alanyl-L-histidinato zinc on cell proliferation is dependent on protein synthesis in osteoblastic MC3T3-E1 cells. Mol Cell Biochem 122: 59–64, 1993
Sudo H, Kodama H, Amagai Y, Yamamoto S, Kasai S:In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol 96: 191–198, 1988
Walter K, Schutt C: Acid and alkaline phosphatase in serum. In: Bergmeyer HU (ed.) Methods in enzymatic analysis. Verlag Chemie Weinheim Academic Press, New York, 1974, vol 1–2, p 856–860
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951
Flanagan B, Nichols G Jr: Metabolic studies of bonein vitro. IV: Collagen biosynthesis by surviving bone fragmentsin vitro. J Biol Chem 237: 3786–3792, 1962
Ceriotti G: Determination of nuclei acids in animal tissues. J Biol Chem 214: 39–77, 1955
Bernier SM, Goltzman D: Effect of protein and steroidal osteotropic agents on differentiation and epidermal growth, factormediated growth of the CFK1 osseous cell line. J Cell Physiol 152: 317–327, 1992
Stein GS, Lian JB, Owen TA: Relationship of cell growth to the regulation of tissue-specific gene expression during osteoblast differentiation. FASEB J 4: 3111–3123, 1990
Parisi AF, Vallee BL: Zihc metalloenzyme: characteristics and significance in biology and medicine. Am J Clin Nutr 22: 1222–1230, 1969
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hashizume, M., Yamaguchi, M. Effect of β-alanyl-L-histidinato zinc on differentiation of osteoblastic MC3T3-El cells: Increases in alkaline phosphatase activity and protein concentration. Mol Cell Biochem 131, 19–24 (1994). https://doi.org/10.1007/BF01075720
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01075720