Yamaguchi M, Hashizume M
Laboratory of Metabolism and Endocrinology, Graduate School of Nutritional Sciences, University of Shizuoka, Japan.
Mol Cell Biochem. 1994 Jul 27;136(2):163-9. doi: 10.1007/BF00926077.
The effect of beta-alanyl-L-histidinato zinc (AHZ) on protein components in osteoblastic MC3T3-E1 cells was investigated. Cells were cultured for 3 days at 37 degrees C in CO2 incubator in plastic dishes containing alpha-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 various concentrations of AHZ or other reagents, and the cells were cultured further 3 or 6 days. The homgenate of cells was analyzed with SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The presence of AHZ (10(-7) to 10(-5) M) caused an appreciable increase of many protein components in cells. Especially, the 67 killo-dalton (kDa) and 44 kDa proteins which are the major components from control cells were clearly increased by the presence of AHZ. Furthermore, the concentrations of osteocalcin, insulin-like growth factor-I and transforming growth factor-beta in the culture medium secreted from osteoblastic cells were markedly increased by the presence of AHZ (10(-6) and 10(-5) M). The effect of AHZ was a greater than that of zinc sulfate (10(-6) and 10(-5) M). The present findings suggest that AHZ can increase many proteins which are involved in the stimulation of bone formation and cell proliferation in osteoblastic cells.
研究了β-丙氨酰-L-组氨酸锌（AHZ）对成骨细胞MC3T3-E1细胞中蛋白质成分的影响。细胞在含有α-改良最小必需培养基并补充10%胎牛血清的塑料培养皿中，于37℃的二氧化碳培养箱中培养3天。培养后，将培养基换成含有0.1%牛血清白蛋白以及不同浓度AHZ或其他试剂的培养基，细胞再进一步培养3天或6天。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析细胞匀浆。AHZ（10⁻⁷至10⁻⁵M）的存在使细胞中许多蛋白质成分明显增加。特别是，对照细胞中的主要成分67千道尔顿（kDa）和44 kDa蛋白质因AHZ的存在而明显增加。此外，成骨细胞分泌的培养基中骨钙素、胰岛素样生长因子-I和转化生长因子-β的浓度因AHZ（10⁻⁶和10⁻⁵M）的存在而显著增加。AHZ的作用大于硫酸锌（10⁻⁶和10⁻⁵M）。目前的研究结果表明，AHZ可以增加许多参与刺激成骨细胞骨形成和细胞增殖的蛋白质。
