Hashizume M, Yamaguchi M
Laboratory of Metabolism and Endocrinology, Graduate School of Nutritional Sciences, University of Shizuoka, Japan.
Mol Cell Biochem. 1993 May 12;122(1):59-64. doi: 10.1007/BF00925737.
The effect of beta-alanyl-L-histidinato zinc (AHZ) on bone metabolism was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37 degrees C in a 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 for appropriate periods of time. The presence of AHZ (10(-7)-10(-5) M) stimulated the proliferation of cells. AHZ (10(-6) and 10(-5) M) increased deoxyribonucleic acid (DNA) content in the cells with 48 hr-culture. This increase was completely blocked by the presence of cycloheximide (10(-6) M) or hydroxyurea (10(-3) M). Also, the presence of cycloheximide (10(-6) M) completely inhibited the AHZ (10(-5) M)-induced increase in the proliferation of cells. Meanwhile, parathyroid hormone (10(-7) M), estrogen (10(-9) M) and insulin (10(-8) M) significantly increased cellular DNA content. However, these hormonal effects clearly lowered in comparison with that of AHZ (10(-5) M). Dibutyryl cyclic AMP (10(-4) M) and zinc sulfate (10(-5) M) did not cause a significant increase in cellular DNA content. The present results support the view that AHZ has a direct specific proliferative effect on osteoblastic cells in vitro and that this effect is dependent on protein synthesis.
在成骨细胞MC3T3-E1细胞中研究了β-丙氨酰-L-组氨酸锌（AHZ）对骨代谢的影响。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，并将细胞进一步培养适当的时间。AHZ（10⁻⁷ - 10⁻⁵ M）的存在刺激了细胞的增殖。AHZ（10⁻⁶和10⁻⁵ M）在48小时培养后增加了细胞中的脱氧核糖核酸（DNA）含量。这种增加被放线菌酮（10⁻⁶ M）或羟基脲（10⁻³ M）的存在完全阻断。此外，放线菌酮（10⁻⁶ M）的存在完全抑制了AHZ（10⁻⁵ M）诱导的细胞增殖增加。同时，甲状旁腺激素（10⁻⁷ M）、雌激素（10⁻⁹ M）和胰岛素（10⁻⁸ M）显著增加了细胞DNA含量。然而，与AHZ（10⁻⁵ M）相比，这些激素的作用明显降低。二丁酰环磷腺苷（10⁻⁴ M）和硫酸锌（10⁻⁵ M）并未导致细胞DNA含量显著增加。目前的结果支持这样的观点，即AHZ在体外对成骨细胞具有直接的特异性增殖作用，且这种作用依赖于蛋白质合成。
