三七皂苷R1-低相对分子质量三七多糖纳米粒的制备及其对SH-SY5Y细胞损伤的保护作用  

Preparation of notoginsenoside R-low molecular weight Panax notoginseng polysaccharide nanoparticles and its protective effect on SH-SY5Y cell injury

作　　者：张梦雪 王瑞莹 李楠 吴晨阳 郑婷婷 邹佩志 韩杰 张运[1] 黎迎[1] 郭一飞[1,2] 孙桂波 董政起[1,2] ZHANG Meng-xue;WANG Rui-ying;LI Nan;WU Chen-yang;ZHENG Ting-ting;ZOU Pei-zhi;HAN Jie;ZHANG Yun;LI Ying;GUO Yi-fei;SUN Gui-bo;DONG Zheng-qi(Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193,China;Key Laboratory for New Drug Discovery Based on Classic Prescriptions,Chinese Academy of Medical Sciences,Beijing 100193,China)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]中国医学科学院基于经典名方的新药发现重点实验室,北京100193

出　　处：《中草药》2021年第10期2918-2926,共9页Chinese Traditional and Herbal Drugs

基　　金：中央高校基本科研业务费专项北京协和医学院创新项目(2018PT35030);中国医学科学院医学与健康科技创新工程(2020-I2M-2-011)。

摘　　要：目的利用水溶性低相对分子质量三七多糖(low molecular weight Panax notoginseng polysaccharide,PNP-L)作为新型载体制备纳米粒,实现对三七皂苷R_(1)的增溶并提高其治疗效果。方法利用SephadexG-100凝胶对三七多糖(Panax notoginseng polysaccharide,PNP)分级纯化,得到相对均质的低相对分子质量三七多糖(PNP-L)。以PNP、PNP-L作为三七皂苷R_(1)的载体,采用乳化溶剂挥发法制备三七皂苷R_(1)-三七多糖(R_(1)-PNP)纳米粒和三七皂苷R_(1)-低相对分子质量三七多糖(R_(1)-PNP-L)纳米粒。测定其载药量及包封率,探索低相对分子质量多糖作为载体的可行性。采用马尔文粒度分析仪对纳米粒粒径、粒度分布及Zeta电位进行测定,差示扫描量热仪(DSC)进行晶型分析,透射电子显微镜(TEM)测定其形态。体外培养人神经母细胞瘤SH-SY5Y细胞,构建缺氧/复氧模型,比较三七皂苷R_(1)、R_(1)-PNP-L物理混合制剂、R_(1)-PNP-L纳米粒对神经细胞损伤的保护作用。结果R_(1)-PNP-L纳米粒平均粒径(139.17±7.22)nm,多分散指数(PDI)为0.41±0.19,Zeta电位(-19.14±3.14)mV,载药量28.62%,包封率85.85%,将三七皂苷R_(1)溶解度提高了32.30倍,并起到缓释作用。R_(1)-PNP-L纳米粒对SH-SY5Y细胞无明显细胞毒性,对缺氧/复氧模型诱导的SH-SY5Y细胞损伤的保护效果R_(1)-PNP-L纳米粒>R_(1)-PNP-L物理混合制剂>三七皂苷R_(1)。结论PNP-L可作为三七皂苷R_(1)的一种良好载体,形成的纳米粒可有效提高三七皂苷R_(1)溶解度并对缺氧/复氧模型诱导的SH-SY5Y细胞损伤具有保护作用。Objective To increase the solubility of notoginsenoside R_(1)and improve the therapeutic effect,hydrophilic low molecular weight Panax notoginseng polysaccharide(PNP-L)was used as a new carrier to construct the nanoparticles.Methods Sephadex G-100 gel was used to fractionate and purify P.notoginseng polysaccharide(PNP)to obtain uniform PNP-L with low molecular weight.Using PNP and PNP-L as the carrier of notoginsenoside R_(1),the notoginsenoside R_(1)-notoginseng polysaccharide nanoparticles(R_(1)-PNP nanoparticles)and notoginsenoside R_(1)-notoginseng polysaccharide nanoparticles with low molecular weight(R_(1)-PNP-L nanoparticles)were prepared by solvent evaporation method.Determining the drug loading and encapsulation efficiency of nanoparticles,and exploring the feasibility of polysaccharides with low molecular weight as carriers.Malvern particle size analyzer was used to measure the particle size,poly dispersity index(PDI)and Zeta potential.Differential scanning calorimeter(DSC)was used to analyze the crystalline form of the nanoparticles.Transmission electron microscopy was used to observe the surface morphology.SH-SY5 Y nerve cells were cultured in vitro and a hypoxia/reoxygenation model was constructed to compare the protection effect of P.notosaponin R_(1),R_(1)-PNP-L physical mixture and R_(1)-PNP-L nanoparticles.Results The particle size,PDI,Zeta potential,drug loading and encapsulation efficiency of R_(1)-PNP-L nanoparticles were(139.17±7.22)nm,0.41±0.19,(-19.14±3.14)mV,28.62%and 85.85%,respectively.R_(1)-PNP-L nanoparticles increased the solubility of notoginsenoside R_(1)by 32.30 times and had slow-release effect.R_(1)-PNP-L nanoparticles had no cytotoxicity to SH-SY5 Y cells.Protection effect on SH-SY5 Y cell injury induced by hypoxia/reoxygenation model:R_(1)-PNP-L nanoparticles>R_(1)-PNP-L physical mixture>notoginsenoside R_(1).Conclusion PNP-L is a good carrier to deliver notoginsenoside R_(1),and the prepared nanoparticles can improve the solubility of notoginsenoside R_(1)effectively and in

关 键 词：三七皂苷R1 三七多糖 新型载体 纳米粒 提高溶解度 SH-SY5Y细胞 损伤保护 乳化溶剂挥发法 缓释 

分 类 号：R283.6[医药卫生—中药学]