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

作　　者：任艳[1] 李萍萍[1] 

机构地区：[1]北京大学临床肿瘤学院北京肿瘤医院暨北京市肿瘤防治研究所中西医结合科恶性肿瘤发病机制及转化研究教育部重点实验室,100142

出　　处：《中国实验方剂学杂志》2010年第13期141-144,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：北京市中医局科技项目(2004京中重IV15)

摘　　要：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.

关 键 词：舒肝健骨方 成骨样细胞MG-63 碱性磷酸酶 雌激素活性 

分 类 号：R285.5[医药卫生—中药学]