淫羊藿总黄酮胶囊活性成分宝藿苷Ⅱ对MC3T3-E1细胞增殖分化的影响  

Effects of baohuoside Ⅱ on the osteoblastic proliferation and differentiation of MC3T3-E1 cells

作　　者：李芳[1,2] 徐忠坤[1,2] 顾少莉 秦建平[1,2] 周军[1,2] 王振中[1,2] 萧伟[1,2] 

机构地区：[1]江苏康缘药业股份有限公司 [2]中药制药过程新技术国家重点实验室,连云港222001

出　　处：《中药药理与临床》2017年第6期54-57,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：科技部十二五"重大新药创制";项目编号:2013ZX09402203

摘　　要：目的:从淫羊藿总黄酮胶囊中制备分离宝藿苷Ⅱ,检测不同浓度宝藿苷II对体外培养的MC3T3-E1细胞增殖分化的影响。方法:分别用10、20、40nmol/ml的宝藿苷II作用MC3T3-E1细胞,CCK-8法检测细胞存活率;比色法测定碱性磷酸酶(ALP)活力;ELISA法检测骨钙素(OT)含量;茜素红染色分析并用氯化十六烷基吡啶测定钙沉积量;Western Blot法检测宝藿苷Ⅱ对BMP2信号通路蛋白表达的影响。结果:10、20、40nmol/ml的宝藿苷Ⅱ均可明显促进MC3T3-E1的增殖;提高MC3T3-E1分泌ALP和OT的能力;矿化结节数量明显增多且钙沉积量吸光度值显著增大;明显提高BMP2、Runx2和Osterix蛋白表达量,且呈剂量依赖性。结论:淫羊藿总黄酮胶囊活性成分宝藿苷II可以明显促进MC3T3-E1细胞的增殖分化。Objective： To prepare and isolate the baohuosideⅡ from the Epimedium flavonoids capsule as well as to investigate the effect of different concentrations of baohuosideⅡ on osteogenic differentiation of cultured MC3 T3-E1 cells in vitro. Methods： MC3 T3-E1 cells were cultured with 10 nmol/ml,20 nmol/ml and 40 nmol/ml of baohuosideⅡ. The cell suivival rate was determined in CCK-8 assay and the alkaline phosphatase（ ALP） activity was detected by colorimetric method. ELISA method was used to detect Osteocalcin（ OT） content. Alizarin red staining and cetylpyridinium chloride were used to analyze and determine the calcium deposition,respectively. Additionally,the effect of baohuosideⅡon proteins in BMP2 signaling pathway was analyzed by Western Blot. Results： 10 nmol/ml,20 nmol/ml and 40 nmol/ml BaohuosideⅡ significantly promoted the proliferation of MC3 T3-E1,enhanced the ability of MC3 T3-E1 to secrete ALP and OT,increased the number of mineralized nodules and the absorbance value of the calcium deposition,as well as obviously improved expressions of BMP2,Runx2 and Osterix protein in a dose-dependent manner. Conclusion： BaohuosideⅡ which comes from Epimedium flavonoids capsule could obviously promote the osteoblastic proliferation and differentiation of MC3 T3-E1 cells.

关 键 词：宝藿苷Ⅱ MC3T3-E1细胞 增殖 分化 

分 类 号：R285[医药卫生—中药学]