Yamaguchi M, Miwa H
Department of Environmental Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Japan.
Pharmacology. 1991;42(4):230-40. doi: 10.1159/000138802.
The present investigation was undertaken to clarify the in vitro effect of beta-alanyl-L-histidinato zinc (AHZ) on bone metabolism in tissue culture. Calvaria were removed from weanling rats (3-week-old male) and cultured for periods up to 96 h in Dulbecco's modified Eagle medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The experimental cultures contained 10(-8) to 10(-4) mol/l AHZ. The bone cellular zinc content was significantly increased in cultures with concentrations of AHZ greater than 10(-6) mol/l. With 10(-5) mol/l zinc sulfate, the bone cellular zinc content was significantly elevated. Bone calcium content was significantly increased by the presence of 10(-7) to 10(-4) mol/l AHZ. This increase was blocked by the presence of 10(-7) mol/l cycloheximide. Bone alkaline phosphatase activity was elevated in the presence of AHZ (10(-7) to 10(-4) mol/l), whereas it did not significantly alter acid phosphatase activity Bone collagen and DNA contents were significantly increased by 10(-7) to 10(-5) mol/l AHZ, while they were not significantly elevated by zinc sulfate (10(-7) and 10(-6) mol/l). The AHZ (10(-5) mol/l)-induced increase in bone alkaline phosphatase activity and DNA content were prevented by 10(-4) mol/l dipicolinate, a chelator of zinc. Furthermore, the AHZ (10(-5) mol/l)-induced increase in bone alkaline phosphatase activity, collagen and DNA contents were blocked by 10(-7) mol/l cycloheximide. These findings indicate that AHZ had a direct stimulatory effect on bone mineralization in vitro, and that bone protein synthesis was a necessary component of this response. The AHZ effect was more intensive than that of zinc sulfate.
本研究旨在阐明β-丙氨酰-L-组氨酸锌（AHZ）在组织培养中对骨代谢的体外作用。从断奶大鼠（3周龄雄性）取出颅骨，在补充了抗生素和牛血清白蛋白的杜尔贝科改良伊格尔培养基（高糖，4.5%）中培养长达96小时。实验培养物中含有10^(-8)至10^(-4)mol/L的AHZ。当AHZ浓度大于10^(-6)mol/L时，骨细胞锌含量显著增加。使用10^(-5)mol/L硫酸锌时，骨细胞锌含量也显著升高。10^(-7)至10^(-4)mol/L的AHZ可使骨钙含量显著增加。这种增加被10^(-7)mol/L的环己酰亚胺所阻断。在AHZ（10^(-7)至10^(-4)mol/L）存在的情况下，骨碱性磷酸酶活性升高，而酸性磷酸酶活性没有显著改变。10^(-7)至10^(-5)mol/L的AHZ可使骨胶原蛋白和DNA含量显著增加，而10^(-7)和10^(-6)mol/L的硫酸锌则未使其显著升高。10^(-4)mol/L的吡啶二羧酸（一种锌螯合剂）可阻止AHZ（10^(-5)mol/L）诱导的骨碱性磷酸酶活性和DNA含量增加。此外，10^(-7)mol/L的环己酰亚胺可阻断AHZ（10^(-5)mol/L）诱导的骨碱性磷酸酶活性、胶原蛋白和DNA含量增加。这些发现表明，AHZ在体外对骨矿化有直接刺激作用，并且骨蛋白合成是该反应的必要组成部分。AHZ的作用比硫酸锌更强烈。
