成骨样细胞体外培养法筛选杜仲叶防治骨质疏松症的药效成份

目的 探讨杜仲叶提取成份对体外培养的成骨样细胞代谢功能的影响,筛选杜仲叶防治骨质疏松的药效成份。方法 从杜仲叶提取分离出4个化合物,以不同浓度(10-4g/L、10-6g/L、10-8g/L)加入体外培养的MG-63成骨细胞株中,并设空白组和对照组。培养4 d后,MTT比色法测定细胞增殖,对硝基磷酸盐法测定培养基中ALP活性。结果 细胞增殖数量与对照组比较,成份I 10-6g/L浓度组、Ⅱ、Ⅲ10-8g/L浓度组和成份Ⅳ10-4g/L、Ⅲ10-8g/L浓度组显著提高(P<0.05);成份Ⅱ、Ⅲ的10-4g/L、10-6g/L浓度组和成份Ⅳ10-6g/L浓度组极显著提高(P<0.01)。ALP活性与对照组比较,成份I 10-6g/L浓度组、成份Ⅱ10-4g/L、10-8g/L浓度组、成份Ⅲ10-8g/L浓度组和成份Ⅳ的10-4g/L、10-8g/L浓度组有显著性增强;成份Ⅱ10-6g/L浓度组和成份Ⅲ的10-4g/L、10-6g/L浓度组均极显著增强(P<0.01)。结论 4个成份均有一定的促进成骨细胞增殖、调节成骨细胞代谢的作用,以Ⅱ和Ⅲ效果明显。

Osteoblast like cell cultured in vitro to screen out effective components of preventing and treating osteoporosis in Duzhong leaf

Objective To explore active components from Duzhong leaves affecting MG-63 osteoblast cultured in vitro, screen out the effective components for preventing and treating osteoporosis in Duzhong leaf. Methods Components(Ⅰ、Ⅱ、Ⅲ、Ⅳ) abstracting from Duzhong leaves with different concentration(10-4g/L、10-6g/Ls、10-8g/L) added to cultured MG-63 osteoblast, then cell proliferation was tested by MTT colorimetry; ALP vigor in culture medium was tested by p-nitroghosphate method. Results Cell quantity in component I with 10-6g/ L, Ⅱ, Ⅲ with 10-6g/L, Ⅳ with 10-4g/L and 10-6g/L concentration groups was increased than that in control group(P < 0.05). Cell quantity in compound Ⅱ , Ⅲ with 10-4g/L and 10-6 g/L groups,Ⅳ with 10-4g/L concentration groups was notably higher than that in control group(P< 0.01). ALP vigor in componentⅠ with 10-4g/L, Ⅱ with 10-4g/L and 10-6g/L, Ⅲ& with 10-8g/L and Ⅳ with 10-4g/ L and 10-6g/L concentration groups was higher than that in control group(P < 0.05) ;component Ⅱwith 10-6g/L and Ⅲ with 10-4g/L and 10 - 6g/L concentration groups was significantly higher than that in the control group(P < 0.01) . Conclusion All components Ⅰ , Ⅱ , Ⅲand Ⅳ can improve osteoblast proliferation and regulate bone metabolism, component Ⅱ and Ⅲare better.