摘要
目的优化蒙药加味塔布森-2(MT-2)的提取工艺,并考察按最优工艺所制提取物对破骨细胞分化的抑制作用。方法采用网络药理学方法筛选MT-2工艺优化的指标性成分;以上述指标性成分的度值和含量计算综合评分,在单因素实验的基础上,使用Box-Behnken设计-响应面法优化MT-2的提取工艺并验证。以核因子κB受体激活蛋白配体(100 ng/mL)诱导RAW264.7细胞以制备破骨细胞分化模型,考察按最优工艺所制MT-2提取物(18.6、37.2、74.4 ng/mL)对破骨细胞分化的抑制作用。结果网络药理学筛选所得指标性成分包括绿原酸、松脂醇二葡萄糖苷、异绿原酸A、1,5-二咖啡酰奎宁酸、羟基红花黄色素A、人参皂苷Rg_(1)、人参皂苷Rb_(1)。MT-2最优提取工艺为乙醇体积分数60%,料液比1∶14(g/mL),提取时间94 min,提取2次;3次验证实验所得平均综合评分为95.50,与预测值(95.75)的相对误差为-0.26%。与破骨分化模型细胞比较,经最优工艺所制MT-2提取物处理后的细胞多为单核圆形细胞,破骨细胞数量均显著减少(P<0.05),且这种抑制作用有随药物质量浓度增加而增强的趋势。结论优化的MT-2提取工艺稳定、可行;所得提取物可抑制破骨细胞分化。
OBJECTIVE To optimize the extraction technology of modified Tabusen-2(MT-2),and to investigate inhibitory effects of the extract obtained by the optimal technology on osteoclast differentiation.METHODS The index components of MT-2 process optimization were selected by using network pharmacology.Based on single factor tests,the extraction technology of MT-2 was optimized by Box-Behnken design-response surface methodology according to the comprehensive score of contents of above index components,and then validated.RAW264.7 cells were induced by receptor activator of nuclear factor-κB ligand(100 ng/mL)to prepare osteoclast differentiation model.Inhibitory effects of MT-2 extract(18.6,37.2,74.4 ng/mL)obtained by the optimal technology on osteoclast differentiation were investigated.RESULTS The index components screened by network pharmacology included chlorogenic acid,terpineol diglucoside,isochlorogenic acid A,1,5-dicaffeoylquinic acid,hydroxysafflower yellow A,ginsenoside Rg_(1) and ginsenoside Rb_(1).The optimal extraction technology of MT-2 was ethanol volume fraction of 60%,the solid-liquid ratio of 1∶14(g/mL),extraction time of 94 min and extraction times of twice.The average comprehensive score obtained by the three validation experiments was 95.50,and the relative error with the predicted value(95.75)was-0.26%.Compared with osteoclastic differentiation model cells,the cells treated with MT-2 extract prepared by the optimal technology were mostly mononuclear round cells,and the number of osteoclasts decreased significantly(P<0.05),its inhibitory effects tended to strengthen with the increase of drug concentration.CONCLUSIONS The optimal extraction technology of MT-2 is stable and feasible.Obtained extract can inhibit osteoclast differentiation.
作者
郭姝
宋清翔
董睿
封千喜
薛培凤
董馨
GUO Shu;SONG Qingxiang;DONG Rui;FENG Qianxi;XUE Peifeng;DONG Xin(School of Pharmacy,Inner Mongolia Medical University,Hohhot 010110,China)
出处
《中国药房》
CAS
北大核心
2022年第16期1943-1949,共7页
China Pharmacy
基金
国家自然科学基金资助项目(No.81960758)
国家大学生创新创业训练计划项目(No.202010132003)
内蒙古自治区大学生创新创业训练计划项目(No.202110132029)。
