增骨Ⅰ、Ⅱ、Ⅲ号对体外培养成骨细胞影响的实验研究

Experimental Study on Effect of Zenggu-Ⅰ, -Ⅱ, -Ⅲ on Osteoblast in Vitro

目的 :观察增骨Ⅰ、Ⅱ、Ⅲ号对体外培养成骨细胞增殖和成骨功能的影响。方法 :利用分段酶消化法从胎鼠颅骨中分离出成骨细胞 ,倒置相差显微镜观察其形态 ,碱性磷酸酶 (ALP)染色来鉴定细胞。然后用不同浓度的增骨Ⅰ、Ⅱ、Ⅲ号作用于成骨细胞 ,使用四甲基偶氮唑蓝 (MTT)比色分析、骨钙素 (BGP)含量放免测定、ALP含量测定、细胞钙含量测定来检测其对成骨细胞增殖和成骨能力的影响。结果 :增骨Ⅲ号以剂量依赖方式促进各项指标的表达 ,促进成骨细胞的增殖及骨形成 ,增骨Ⅰ、Ⅱ号对体外培养成骨细胞无影响。结论 :增骨Ⅲ号促进成骨细胞的增殖 ,促进骨形成 ,增骨Ⅰ、Ⅱ、Ⅲ号序贯疗法可有效防治骨质疏松症和促进骨形成。

Objective: To investigate the effect of Zenggu (ZG) Ⅰ, Ⅱ and Ⅲ on osteoblast (OB) cultured in vitro. Methods: OB was isolated from calvaria of 22 day old fetal SD rats by means of modified sequential collagenase digestion and incubated in RM1640 medium and the cell morphology was observed by alkaline phosphatase (ALP) staining. Different concentrations of ZG Ⅰ, Ⅱ and Ⅲ were added to the OB and incubated. The effect of ZG on the proliferation and osteogenesis of OB was monitored by MTT analysis, BGP content radioimmunoassay, ALP activity and calcium contents determination. Results: The BGP, ALP, and calcium contents of the cultured OB cells as well as the cell proliferation were higher in ZG Ⅲ group than those in the other groups (P<0 05) in a dose dependent manner. It suggested that ZG Ⅲ could promote the proliferation and osteogenesis of OB, but ZG Ⅰ and ZG Ⅱ had insignificant influence on OB cultured in vitro. Conclusion: ZG Ⅲ could promote the OB proliferation and bone formation, ZG Ⅰ, Ⅱ, Ⅲ sequential therapy could effectively prevent and treat osteoporosis and promote the bone formation.