Uchiyama Satoshi, Yamaguchi Masayoshi
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Shizuoka 422-8526, Japan.
Int J Mol Med. 2005 Apr;15(4):675-81.
The carotenoid beta-cryptoxanthin has been shown to have a stimulatory effect on bone formation in rat bone tissues in vitro. The effect of beta-cryptoxanthin in osteoblastic cells in vitro was investigated. Osteoblastic MC3T3-E1 cells were cultured for 72 h in alpha-minimal essential medium containing 10% fetal bovine sereum (FBS) to reach subconfluent monolayers. After culture, the medium was changed, then beta-cryptoxanthin (10(-8) to 10(-6) M) was added in the culture medium without FBS, and the cells were cultured for an additional 24, 48, or 72 h. The proliferation of osteoblastic cells was significantly enhanced in the presence of beta-cryptoxanthin (10(-8) to 10(-6) M), when it was cultured for 48 or 72 h in medium containing 10% FBS. When osteoblastic cells with subconfluency were cultured for 48 or 72 h in FBS free-medium containing beta-cryptoxanthin (10(-8) to 10(-6) M), alkaline phosphatase activity or deoxyribonucleic acid (DNA) content in the cells was significantly increased. Also, protein content in the cells was significantly increased by culture with 10(-6) M beta-cryptoxanthin for 48 or 72 h. The effect of beta-cryptoxanthin (10(-6) M) in increasing protein content, alkaline phosphatase activity, or DNA content in the cells was significantly blocked in the presence of staurosporine (10(-6) M) or PD98059 (10(-6) M), which is an inhibitor of protein kinases. The stimulatory effect of beta-cryptoxanthin (10(-6) M) on cellular biochemical components was completely prevented in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis, or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB; 10(-9) M), an inhibitor of transcriptional activity. The expressions of insulin-like growth factor (IGF)-I and transforming growth factor (TGF)-beta1 mRNAs were demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis in osteoblastic cells using mouse IGF-I or TGF-beta1-specific primers. These expressions were significantly raised in the presence of beta-cryptoxanthin (10(-6) M). This study demonstrates that beta-cryptoxanthin has a stimulatory effect on cell proliferation and biochemical components in osteoclastic MC3T3-E1 cells, and that the carotenoid can stimulate transcriptional activity in the cells.
类胡萝卜素β-隐黄质已被证明在体外对大鼠骨组织的骨形成具有刺激作用。研究了β-隐黄质在体外对成骨细胞的影响。将成骨MC3T3-E1细胞在含有10%胎牛血清（FBS）的α-最低必需培养基中培养72小时，以达到亚汇合单层。培养后，更换培养基，然后在不含FBS的培养基中加入β-隐黄质（10⁻⁸至10⁻⁶M），并将细胞再培养24、48或72小时。当在含有10%FBS的培养基中培养48或72小时时，β-隐黄质（10⁻⁸至10⁻⁶M）存在下成骨细胞的增殖显著增强。当亚汇合的成骨细胞在含有β-隐黄质（10⁻⁸至10⁻⁶M）的无FBS培养基中培养48或72小时时，细胞中的碱性磷酸酶活性或脱氧核糖核酸（DNA）含量显著增加。此外，用10⁻⁶Mβ-隐黄质培养48或72小时，细胞中的蛋白质含量也显著增加。在存在星形孢菌素（10⁻⁶M）或PD98059（10⁻⁶M，一种蛋白激酶抑制剂）的情况下，β-隐黄质（10⁻⁶M）增加细胞中蛋白质含量、碱性磷酸酶活性或DNA含量的作用被显著阻断。在存在蛋白质合成抑制剂环己酰亚胺（10⁻⁶M）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；10⁻⁹M）的情况下，β-隐黄质（10⁻⁶M）对细胞生化成分的刺激作用被完全阻止。使用小鼠IGF-I或TGF-β1特异性引物，通过逆转录-聚合酶链反应（RT-PCR）分析在成骨细胞中证实了胰岛素样生长因子（IGF）-I和转化生长因子（TGF）-β1 mRNA的表达。在β-隐黄质（10⁻⁶M）存在下，这些表达显著升高。本研究表明，β-隐黄质对破骨MC3T3-E1细胞的细胞增殖和生化成分具有刺激作用，并且该类胡萝卜素可以刺激细胞中的转录活性。
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