加味塔布森-2提取工艺优化及其对破骨细胞分化的抑制作用
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篇名: 加味塔布森-2提取工艺优化及其对破骨细胞分化的抑制作用
TITLE: Optimization of the extraction technology of modified Tabusen- 2 and its inhibitory effects on osteoclast differen- tiation
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(100ng/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、74.4ng/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Rg1、人参皂苷Rb1。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94min,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。
ABSTRACT: 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. RAW 264.7 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.
期刊: 2022年第33卷第16期
作者: 郭姝,宋清翔,董睿,封千喜,薛培凤,董馨
AUTHORS: GUO Shu,SONG Qingxiang,DONG Rui,FENG Qianxi,XUE Peifeng,DONG Xin
关键字: 加味塔布森-2;蒙药;提取工艺;Box-Behnken设计-响应面法;破骨细胞分化;抑制作用
KEYWORDS: modified Tarbson- 2; Mongolian medicine ; extraction technology ; Box-Behnken design-response surface
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