舒肝健骨方对成骨样细胞增殖分化的影响及雌激素活性的检测

Effects of Shugan Jiangu Decoction on Osteoblast-Like Cells Proliferation,Differentiation and Evaluation of Estrogenic Potential of Decoction

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 investigate the effects of Shugan Jiangu decoction on osteoblast-like cells and evaluate the estrogenic potential of the decoction.Method： Human osteoblast-like cells MG-63 were cultured and treated with Shugan Jiangu decoction.Cell proliferation was investigated by MTT assay.Alkaline phosphatase activity,as indicator of cell differentiation,was assayed by p-nitrophenylphosphate（pNPP） method.After transient cotransfection with plasmid pERE-TK-Luc and pRL-TK in MCF-7 cells,we evaluated the estrogenic potential of Shugan Jiangu decoction by dual-luciferase reporter assay（DLR）.Result： By MTT assay,at 48,72 h,Shugan Jiangu decoction（0.75-12 g·L^-1） increased osteoblast-like cell proliferation dose-dependently.By pNPP method assay,at 24,48,72 h,the decoction enhanced alkaline phosphatase activity dose-dependently.By DLR assay,there was no significant difference in estrogenic potential between Shugan Jiangu decoction and negative control.Conclusion： Shugan Jiangu decoction stimulated osteoblast-like cells proliferation and differentiation.No estrogenic potential was observed for Shugan Jiangu decoction.