目的 研究塞隆骨提取物对体外培养大鼠成骨样细胞ROS17/2.8代谢功能的调节作用,从而在细胞水平上阐述塞隆骨防治骨质疏松的作用机制及药效成分的筛选。方法 采用超临界萃取法制备塞隆骨的脂、醇、水提液及水煎提取液,用MTT法测定塞隆骨脂、醇、水提液及水煎液对成骨样细胞的增殖作用;测定碱性磷酸酶(ALP)活性,观察上述提取液对细胞分化的影响。结果 10mg/mL塞隆骨脂提液和水提液对大鼠成骨样细胞ROS17/2.8有显著的增殖作用(P<0.01);ALP检测结果显示,10mg/mL脂提液能极显著增强成骨样细胞内ALP活性(P<0.01),1.0、0.1mg/mL能提高ALP活性(P<0.05);10mg/mL水提液能使ALP活性升高(P<0.05)。结论 塞隆骨脂和水提取液中分别存在有较高活性的促成骨样细胞增殖、分化的物质,可作为虎骨的替代中药。

Objective To investigate the modulation of Sailong bone extracts on rat osteoblastoid cells ROS17/2.8 in vitro so as to clarify the mechanism of osteoporosis prevention and active component screening of Salong bone. Methods Sailong bone fat-soluble extract, Sailong bone ethanol extract and Sailong bone aqueous extract were extracted with super critical fluid extraction (SCFE), and Sailong bone boiling-water component was extracted with distilled water directly. MTT assay was applied to determine the proliferation of the cell promoted by four Sailong bone extracts at different doses. Differentiating effects of the extracts with different concentrations on the cell were evaluated by the examinations of alkali phosphate (ALP) activity. Results Sailong bone fat-soluble and aqueous extract (each 10 mg/mL) could significantly improve the proliferation of rat osteoblastoid cells ROS17/2.8 (P0.01). Sailong bone fat-soluble extract (10 mg/mL) significantly enhanced ALP activity (P0.01). Fat-soluble extract (1.0, 0.1 mg/mL) could improve the ALP activitiy (P0.05). Aqueous extract (10 mg/mL) could improve the ALP activity (P0.05). Conclusion In the Sailong bone fat-soluble extract and Sailong bone aqueous extract, there are some components promoting the proliferation and differentiation of osteoblast, and Sailong bone can be the substitute of the bone of tiger as a Chinese materia medica.

四川省科技厅科技攻关项目(01SG003-14)