目的研究云南白药对体外成骨细胞增殖和分化的影响作用,为其促进骨修复的机制提供理论依据。方法取胎鼠颅盖骨,用贴壁组织块反复消化法获取培养成骨细胞,碱性磷酸酶及钙结节染色对其鉴定;用不同质量浓度的云南白药刺激成骨细胞;用甲噻唑四唑氮比色法检测成骨细胞增殖能力,分光光度计和酶联免疫吸附法分别检测细胞裂解液中碱性磷酸酶活性和骨钙蛋白的含量。结果与空白对照组相比,不同药物质量浓度组均可以促进成骨细胞增殖,并且在第5、7d,5mg.L^-1组作用更明显,同时也可增加碱性磷酸酶的活性和促进骨钙蛋白合成。结论云南白药可促进成骨细胞的增殖分化。
Objective To investigate the effects of Yunnan Baiyao on promoting osteoblast proliferation and differentiation, and to provide theoretical basis for its bone repair mechanism. Methods Cranium was harvested from fetal SD rats. Then, osteoblast was acquired and cultured by repeating enzymatic digestion-adherent explant method identified via invert microscope, alkaline phosphorase stain, and stain of calcified nodules. After using different concentrations of Yunnan Baiyao stimulating osteoblasts, the proliferation of the acquired cells was examined through methyl thiazolyl tetrazolium assay. Alkaline phosphatase(ALP) activity and osteocalcin content were analyzed by using a spectrophotometer and enzyme linked immunosorbent assay in the cell lysate liquid. Results Compared with the controls, three different concentration groups can promote the proliferation of osteoblast; the 5 mg'L1 group had more obvious roles in 5 and 7 d. It also increased the activity of ALP and promoted osteocalcin synthesis simultaneously. Conclusion Yunnan Baiyao can promote proliferation and differentiation of osteoblasts.