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目的:观察舒肝健骨方对成骨样细胞增殖分化的影响,并检测舒肝健骨方是否有雌激素活性。方法:培养成骨样细胞MG-63,给予不同浓度的舒肝健骨方处理后,MTT法检测复方对MG-63细胞增殖的影响;对硝基苯酚法检测复方对MG-63细胞碱性磷酸酶(Alkaline phosphatase,ALP)活性的影响。培养MCF-7细胞,转染萤火虫荧光素酶和海肾荧光素酶质粒,双荧光素酶报告基因检测试剂检测复方雌激素活性。结果:舒肝健骨方在处理MG-63细胞48,72 h时,0.75～12 g·L-1的剂量能促进成骨样细胞增殖,并且呈现剂量依赖性;在作用24,48,72 h时,促进成骨样细胞碱性磷酸酶活性,呈剂量依赖性。舒肝健骨方的雌激素活性与阴性对照相比无统计学差异,与阳性对照雌二醇相比有显著性差异(P<0.05)。结论:舒肝健骨方能够促进成骨样细胞的增殖分化,并且无雌激素活性。
Objective: To observe the effects of Shugan Jiangu Fang on proliferation and differentiation of osteoblast-like cells and to detect whether Shugan Jiangu Fang has estrogenic activity. Methods: The osteoblast-like cells MG-63 were cultured and treated with Shugan Jiangu Decoction at various concentrations. MTT assay was used to detect the effect of compound prescription on the proliferation of MG-63 cells. The effect of compound prescription on MG- Alkaline phosphatase (ALP) activity. MCF-7 cells were cultured, firefly luciferase and Renilla luciferase plasmids were transfected, and dual-luciferase reporter assay was used to detect the activity of compound estrogen. Results: Shuganjiangyuke decoction dose-dependently increased the proliferation of osteoblast-like cells at dose of 0.75-12 g · L-1 at 48 and 72 h, h, promote osteoblast-like cell alkaline phosphatase activity in a dose-dependent manner. There was no significant difference in the estrogenic activity between Shugan Jiangu Fang and the negative control, which was significantly different from the positive control estradiol (P <0.05). Conclusion: Shugan Jiangu Fang can promote the proliferation and differentiation of osteoblast-like cells without estrogen activity.