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  • 沈燚,何玉琼,张奇,刘梦琴,辛海量,张泉龙,秦路平,张巧艳.巴戟天环烯醚萜苷类成分的制备及其对破骨细胞骨吸收的抑制作用[J].第二军医大学学报,2019,40(2):149-156    [点击复制]
  • SHEN Yi,HE Yu-qiong,ZHANG Qi,LIU Meng-qin,XIN Hai-liang,ZHANG Quan-long,QIN Lu-ping,ZHANG Qiao-yan.Preparation of Morinda officinalis iridoid glycosides and the inhibitory effect on bone resorption of osteoclasts[J].Acad J Sec Mil Med Univ,2019,40(2):149-156   [点击复制]
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巴戟天环烯醚萜苷类成分的制备及其对破骨细胞骨吸收的抑制作用
沈燚1,2,3,何玉琼1,张奇2,3,刘梦琴2,3,辛海量1,张泉龙3,秦路平1,3*,张巧艳1,2,3*
0
(1. 海军军医大学(第二军医大学)药学院生药学教研室, 上海 200433;
2. 福建中医药大学药学院, 福州 350122;
3. 浙江中医药大学药学院生药学教研室, 杭州 310053
*通信作者)
摘要:
目的 研究巴戟天中环烯醚萜苷类成分(Morinda officinalis iridoid glycosides,MOIG)的制备方法及其对破骨细胞骨吸收的抑制作用。方法 用高效液相色谱(HPLC)法测定MOIG中水晶兰苷和去乙酰基车叶草苷酸的含量;通过对AB-8、ADS-17、D101、HPD400、HPD600、HP20、S-8、SP850、XDA-1、XDA-6等型号大孔吸附树脂进行静态吸附-解吸附实验筛选大孔树脂型号,并优化大孔树脂富集MOIG的条件;以由小鼠骨髓单核细胞诱导的破骨细胞为模型评价MOIG对骨吸收的抑制作用。结果 经过优选,XDA-1型大孔树脂对MOIG有较好的吸附和解吸附作用,优化的XDA-1型大孔树脂富集MOIG的条件为:上样液中MOIG的质量浓度为19.15 mg/mL,pH值为1.0,树脂柱的径高比为1︰7,上样流速为2.0 BV/h(1 BV=80 mL),上样体积为0.75 BV,树脂吸附时间为11 h,7 BV水洗脱以除去其他的成分,10%乙醇以3.0 BV/h流速洗脱MOIG。验证试验结果表明,所制备的MOIG中水晶兰苷和去乙酰基车叶草苷酸的总含量可达54%以上。MOIG对由小鼠骨髓单核细胞诱导的破骨细胞增殖无显著影响,但能抑制破骨细胞抗酒石酸酸性磷酸酶活性,抑制破骨细胞的骨吸收(P<0.05,P<0.01)。结论 XDA-1型大孔树脂可用于富集MOIG,得到的水晶兰苷和去乙酰基车叶草苷酸总含量在54%以上;MOIG可抑制破骨细胞的骨吸收。
关键词:  巴戟天  环烯醚萜苷  大孔树脂  破骨细胞  骨吸收
DOI:10.16781/j.0258-879x.2019.02.0149
投稿时间:2018-09-27修订日期:2018-11-12
基金项目:国家自然科学基金(U1505226).
Preparation of Morinda officinalis iridoid glycosides and the inhibitory effect on bone resorption of osteoclasts
SHEN Yi1,2,3,HE Yu-qiong1,ZHANG Qi2,3,LIU Meng-qin2,3,XIN Hai-liang1,ZHANG Quan-long3,QIN Lu-ping1,3*,ZHANG Qiao-yan1,2,3*
(1. Department of Pharmacognosy, School of Pharmacy, Naval Medical University(Second Military Medical University), Shanghai 200433, China;
2. School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian, China;
3. Department of Pharmacognosy, School of Pharmacy, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China
*Corresponding authors)
Abstract:
Objective To study the preparation method of Morinda officinalis iridoid glycosides (MOIG) and to explore the inhibitory effect on bone resorption of osteoclasts. Methods High-performance liquid chromatography (HPLC) was applied to determine the contents of monotropein and deacetyl asperulosidic acid from MOIG. The static adsorption-desorption test was used to screen the types of macroporous resins of AB-8, ADS-17, D101, HPD400, HPD600, HP20, S-8, SP850, XDA-1 and XDA-6, and to optimize the related parameters in process of enriching MOIG using macroporous resins. Furthermore, the osteoclasts induced from mouse bone marrow monocytes were used to evaluate the inhibitory effects of MOIG on osteoclastic bone resorption. Results After optimization, XDA-1 macroporous resin had better adsorption and desorption effects on MOIG. The optimized preparation conditions were as follows:mass concentration of MOIG in the sample solutions was 19.15 mg/mL, pH value was 1.0, diameter-height ratio of resin column was 1︰7, flow rate of loading samples was 2.0 BV/h (1 BV=80 mL), loading volume was 0.75 BV, and adsorption time was 11 h. 7 BV water was used to remove other constituents, and 10% ethanol was used to elute MOIG in the flow rate of 3.0 BV/h. The total content of monotropein and deacetyl asperulosidic acid in the prepared MOIG was more than 54%. MOIG had no significant effect on proliferation of the osteoclasts induced by mouse bone marrow monocytes, while it could significantly inhibit the tartrate-resistant acid phosphatase activity and the bone resorption of osteoclasts (P<0.05, P<0.01). Conclusion XDA-1 macroporous resin can be used to enrich MOIG, with more than 54% total content of monotropein and deacetyl asperulosidic acid. The MOIG can significantly inhibit the bone resorption of osteoclasts.
Key words:  Morinda officinalis  iridoid glycosides  macroporous resin  osteoclasts  bone resorption