染料木黄酮对骨形成的促进作用研究

目的:用成年人成骨细胞体外培养的方法,观察染料木黄酮(genistein,GS)对骨形成的影响。方法:将生长状态良好的第三代成骨细胞用PBS洗涤后接种于无酚红高糖DMEM培养液中(含5%经活性碳-葡聚糖苷处理的胎牛血清),实验设二甲基亚砜溶剂对照、雌二醇对照(E2)、抗雌激素它莫西芬对照(TAM)及三个GS剂量组,观察指标包括细胞增殖,碱性磷酸酶活性及细胞骨钙蛋白合成量。结果:与溶剂对照组相比,10-8mol/L E2、10-7mol/L GS和10-6mol/L GS对人成骨细胞处理48h,能够明显促进成骨细胞的增殖作用,使细胞骨钙蛋白合成量显著增加,细胞内碱性磷酸酶活性明显提高;同时,GS可保护因雌激素耗尽所造成的成骨细胞内Ⅰ型胶原蛋白降低。10-7mol/L TAM可拮抗GS对成骨细胞所产生的这些生物学效应。结论:GS具有雌激素效应,可模拟E2而刺激骨形成并保护成骨细胞因雌激素缺乏所造成的细胞内Ⅰ型胶原蛋白降低。由于这一效应可被TAM拮抗,此结果提示,GS的促进骨形成作用是通过与雌激素受体结合产生的。

STUDY ON THE PROMOTIVE EFFECT OF GENISTEIN ON BONE FORMATION

Objective: To investigate the promotive effect of genistein (GS) on bone formation using in vitro adult human periosteum osteoblast. Methods: (1) Adult human periosteum osteoblast culture, isolation and identification. (2) The passage 3 osteoblast cells were used for experiment. (3) The cells were maintained in phenol red-free DMEM supplemented with 5% charcoal-stripped fetal calf serum. The project included four groups: three control groups (estradiol, anti-estrogen and vehicle) and three concentrations of GS group. Cell proliferation, activity of alkaline phosphatase (AKP) and osteocalcin were used for bone formation markers. Results: Compared with vehicle control, 10-8 mol/L E2,10-7 mol/L GS and10-6 mol/L GS could significantly increase osteoblast cell proliferation, activity of alkaline phosphatase (AKP) and osteocalcin synthesis. The maximum effect was observed after treatment with 5×10-7 mol/L GS for 24 h or longer. Additionally, GS prevents typeⅠcollagen decrease induced by estrogen depletion in osteoblast. Conclusion: Dietary genistein exerted anabolic effects by stimulating matrix synthesis and cell proliferation in osteoblast and by interacting with estrogen receptors.