Ishida Koichi, Yamaguchi Masayoshi
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Shizuoka 422-8526, Japan.
Int J Mol Med. 2004 Dec;14(6):1077-81.
The role of albumin in osteoblastic cells was investigated. Osteoblastic MC3T3-E1 cells with subconfluent monolayers were cultured for 24 to 72 h in a medium without fetal bovine serum (FBS) in the presence or absence of albumin (0.25, 0.5, or 1.0 mg/ml of medium). The number of osteoblastic cells was significantly increased by culture for 24 to 72 h in the presence of albumin (1.0 mg/ml). The effect of albumin in increasing cell number was completely abolished in the presence of PD98049 (10(-7) M), staurosporine (10(-7) M), or dibucaine (10(-7) M), which is an inhibitor of various protein kinases. Also, the stimulating effect of albumin on cell proliferation was completely prevented in the presence of cycloheximide (10(-7) M), an inhibitor of protein synthesis, or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), an inhibitor of transcriptional activity. DNA content was significantly increased in the cells cultured with albumin (1.0 mg/ml) addition for 24 to 72 h. Alkaline phosphatase activity in the cells, which participates in osteoblastic calcification, was significantly decreased by the culture with albumin (0.25, 0.5, or 1.0 mg/ml) for 24 to 72 h. The expression of insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 mRNAs using transcription-polymerase chain reaction (RT-PCR) analysis in osteoblastic cells was not significantly altered by culture for 48 h with albumin (1.0 mg/ml). This study demonstrated that albumin had a role in the regulation of osteoblastic cell function.
研究了白蛋白在成骨细胞中的作用。将亚汇合单层的成骨MC3T3-E1细胞在无胎牛血清(FBS)的培养基中,于有或无白蛋白(0.25、0.5或1.0mg/ml培养基)存在的情况下培养24至72小时。在存在白蛋白(1.0mg/ml)的情况下培养24至72小时,成骨细胞数量显著增加。在存在PD98049(10^(-7)M)、星形孢菌素(10^(-7)M)或丁卡因(10^(-7)M,一种多种蛋白激酶的抑制剂)时,白蛋白增加细胞数量的作用完全被消除。此外,在存在蛋白质合成抑制剂环己酰亚胺(10^(-7)M)或转录活性抑制剂5,6-二氯-1-β-D-呋喃核糖基苯并咪唑(DRB)时,白蛋白对细胞增殖的刺激作用完全被阻止。添加白蛋白(1.0mg/ml)培养24至72小时的细胞中DNA含量显著增加。参与成骨细胞钙化的细胞中的碱性磷酸酶活性,在使用白蛋白(0.25、0.5或1.0mg/ml)培养24至72小时后显著降低。使用转录-聚合酶链反应(RT-PCR)分析成骨细胞中胰岛素样生长因子-I(IGF-I)和转化生长因子-β1 mRNA的表达,在使用白蛋白(1.0mg/ml)培养48小时后未显著改变。本研究表明白蛋白在成骨细胞功能调节中起作用。