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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 Δ
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郭 姝 ,宋清翔,董 睿,封千喜,薛培凤,董 馨(内蒙古医科大学药学院,呼和浩特 010110)
中图分类号 R965;R283 文献标志码 A 文章编号 1001-0408(2022)16-1943-07
DOI 10.6039/j.issn.1001-0408.2022.16.05
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。
关键词 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
Optimization of the extraction technology of modified Tabusen-2 and its inhibitory effects on osteoclast
differentiation
GUO Shu,SONG Qingxiang,DONG Rui,FENG Qianxi,XUE Peifeng,DONG Xin(School of Pharmacy,Inner
Mongolia Medical University,Hohhot 010110,China)
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. 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 Rg1 and ginsenoside Rb1. 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.
KEYWORDS modified Tarbson-2; Mongolian medicine; extraction technology; Box-Behnken design-response surface
methodology;osteoclast differentiation;inhibitory effect
蒙药加味塔布森-2(modified Tabusen-2,MT-2)是我 材打粉混合而成的散剂,患者用药依从性差,加之方中
国蒙古族治疗骨科疾病的传统验方,由蓝刺头、杜仲、红 各味药材未经提取,不利于有效成分的吸收。复方制剂
花、三七、珍珠5味药材构成,用于治疗骨质疏松、软组织 所含成分复杂,且具有多成分、多靶点协同起效的特
挫伤等,具有良好的临床疗效 [1-2] 。MT-2 原剂型是以药 点 [3-4] 。既往关于复方提取工艺优化的研究大多未能将
Δ 基金项目 国家自然科学基金资助项目(No.81960758);国家大 有效成分含量与药效关联起来,对提取工艺优劣的评价
[5]
学生创新创业训练计划项目(No.202010132003);内蒙古自治区大学 过于片面,造成成分与疗效脱节 。因此,开展 MT-2 成
生创新创业训练计划项目(No.202110132029) 分、疗效、提取工艺的关联研究,对阐明 MT-2 的治疗作
*第一作者 硕士研究生。研究方向:中蒙药物质基础及质量控
用至关重要。为此,本研究拟借助网络药理学方法,挖
制。E-mail:54257065@qq.com
掘MT-2方中的有效成分,并以此作为MT-2提取工艺优
# 通信作者 实验师,硕士生导师,硕士。研究方向:中蒙药物质基
础及质量控制。电话:0471-6653162。E-mail:dongx1128@163.com 劣的评价指标,采用Box-Behnken设计-响应面法考察各
中国药房 2022年第33卷第16期 China Pharmacy 2022 Vol. 33 No. 16 ·1943 ·