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纳米磁珠辅助红细胞膜垂钓技术及其在当归活性成分筛选中的应用 被引量:4

Application of magnetic nanoparticle-assisted erythrocyte membrane fishing assay on active compounds screening from Angelica sinensis
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摘要 目的为提高天然产物中活性成分的筛选效率,利用活性成分与细胞膜(CMs)受体间具有特异性亲和的性质,建立一种新型、高效的纳米磁珠(MNPs)辅助CMs垂钓的方法。方法将MNPs与兔红细胞CMs结合形成纳米磁珠-细胞膜复合物(MN-CMs),利用MN-CMs垂钓当归水提取液中的活性成分,通过GC-MS对垂钓结果进行分析。结果自制MNPs平均粒径为(250.6±3.3)nm(n=3),且粒径分散均匀,多分散指数(PDI)为0.010±0.003(n=3),具有较强的磁性和良好的超顺磁性,饱和磁化强度值为83.4 emu/g。MNPs能与CMs稳定结合,10 mg MNPs的最大吸附量为1.02 mg CMs,同时可以有效保持一定的CMs酶活性。GC-MS结果表明,MN-CMs从当归水提取液中垂钓出的潜在活性成分为藁本内酯,出峰时间为20 min,同时该新型垂钓方法能够有效避免非活性成分的干扰。结论建立的MNPs辅助CMs垂钓方法能够特异性垂钓出当归中活性成分藁本内酯,弥补了传统药物筛选方法的缺点,也为其他天然产物的活性成分筛选提供一种新方法。 Objective To improve the screening efficiency of the active ingredients in natural products by building up a kind of novel and efficient magnetic nanoparticle-assisted cell membranes(MN-CMs) fishing assay employing specific affinity interactions between active ingredients and receptors on cell membranes(CMs). Methods The magnetic nanoparticles(MNPs) were combined with erythrocyte membrane of rabbits to fish active ingredients from water extracts of Angelica sinensis, and the fishing results were analyzed by GC-MS. Results The particle size of the self-made magnetic beads was about(250.6 ± 3.3) nm(n = 3) with PDI index at 0.010 ± 0.003(n = 3), and the beads were monodisperse, strongly magnetic and superparamagnetic, the saturation magnetization was 83.4 emu/g. The combination of MNPs and CMs was stable, the maximum combined amount was 1.02 mg CMs/10 mg MNPs, and the combination was able to keep better enzyme activity of CMs in the fishing assay. GC-MS results showed that ligustilide was fished out as the active compound from water extracts of A. sinensis by MN-CMs assay with the retention time at 20 min, and the new established fishing assay could effectively avoid the interference of inactive components. Conclusion Ligustilide, one of the active ingredients in A. sinensis, can be screened out by the established fishing assay of MN-CMs. The developed fishing method in this workmakes up for some deficiencies of traditional screening method and provides a novel and efficient way to screen ingredients from natural products.
作者 何嘉辉 茆睿智 唐铖 徐亮 肖学凤 HE Jia-hui1, MAO Rui-zhi2, TANG Cheng2, XU Liang2,3, XIAO Xue-feng1(1.School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; 2. Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; 3. Tianjin Medical College, Tianjin 300222, Chin)
出处 《中草药》 CAS CSCD 北大核心 2018年第5期1041-1047,共7页 Chinese Traditional and Herbal Drugs
基金 国家自然科学基金资助项目(81402889) 天津市自然科学基金资助项目(2014JCYBJC24300)
关键词 天然产物 当归 活性成分 细胞膜 纳米磁珠 垂钓 纳米磁珠-细胞膜复合物 磁性 藁本内酯 natural products Angelica sinensis(OLiv.) Diels active ingredients cell membrane magnetic nanoparticle fishing magnetic nanoparticle-assisted cell membranes magnetic ligustilide
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