黄芪多糖-超顺磁性氧化铁纳米复合物的制备及其诱导巨噬细胞极化的活性研究  

Preparation and polarization activity research of astragalus polysaccharide-superparamagnetic iron oxide nanocomposite

作　　者：黄琳清 史新萌 王静蓉[3,4] 瞿鼎 刘玉萍[1,2] 陈彦[1,2] HUANG Lin-qing;SHI Xin-meng;WANG Jing-rong;QU Ding;LIU Yu-ping;CHEN Yan(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,China;Multi-component of Traditional Chinese Medicine and Microecology Research Center,Jiangsu Provincial Academy of Chinese Medicine,Nanjing 210028,China;State Key Laboratory for Quality Research of Chinese Medicines,Macao University of Science and Technology,Macao 999078,China;State Key Laboratory of Dampness Syndrome of Chinese Medicine,The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120,China)

机构地区：[1]南京中医药大学附属中西医结合医院,江苏南京210028 [2]江苏省中医药研究院,中药组分与微生态研究中心,江苏南京210028 [3]中药质量研究国家重点实验室(澳门科技大学),中国澳门999078 [4]广州中医药大学第二附属医院,省部共建中医湿证国家重点实验室,广东广州510120

出　　处：《药学学报》2023年第3期779-788,共10页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82173985,81873017);江苏省卫生健康委医学科研项目(K2019007);中药质量研究国家重点实验室(澳门科技大学)开放课题(2R2104)。

摘　　要：尺寸与表面修饰物是影响超顺磁性氧化铁纳米(superparamagnetic iron oxide nanoparticles,SPIONs)诱导巨噬细胞极化作用的两个关键因素。粒径越小的SPIONs诱导巨噬细胞M1极化效果越好,其次,采用合理的SPIONs表面修饰方法还可达到增强极化效果的目的。本研究使用溶剂热法制备SPIONs并采用Box-Benhnken中心组合实验设计和响应面法优化制备方法,获得小粒径SPIONs。以黄芪多糖(astragalus polysaccharide,APS)为SPIONs亲水性表面修饰物,通过EDC/NHS酯化法成功构建黄芪多糖-超顺磁性氧化铁纳米复合物(APS-SPIONs)。采用动态光散射仪及红外光谱仪对APS-SPIONs的结构进行确证;采用分光光度法对铁、多糖含量进行表征;流式细胞术考察APS-SPIONs对诱导RAW264.7细胞极化的影响;构建RAW264.7巨噬细胞与HepG2人肝癌细胞Transwell共培养体系,初步考察给药后巨噬细胞体外杀伤肿瘤细胞的效果,CCK-8(cell counting kit-8)法考察APSSPIONs对小鼠单核巨噬细胞RAW264.7的增殖活性影响。最终成功制得粒径在(82.93±1.47)nm,zeta电位为(-24.00±0.47)mV的APS-SPIONs。经检测,多糖含量为8.69%,铁含量为7.04%。APS-SPIONs在体外可有效诱导RAW264.7向M1型极化,在共培养体系中提高巨噬细胞对肝癌细胞的杀伤能力,且对巨噬细胞的增殖无抑制作用。本研究为极化巨噬细胞并重塑肿瘤免疫微环境达到杀伤肿瘤作用的药物研发提供策略和方法。Size and surface modification are the two key factors affecting the effect of macrophages polarization induced by superparamagnetic iron oxide nanoparticles(SPIONs).The smaller the particle size,the better the polarization effect of SPIONs.Besides,the reasonable SPIONs surface modification method can also be used to enhance the polarization effect.In this study,SPIONs was prepared by solvothermal method and optimized by Box-Benhnken center combination design and response surface method.Furthermore,astragalus polysaccharidesuperparamagnetic iron oxide nanocomplex(APS-SPIONs)was successfully constructed by EDC/NHS esterification method.The structure of APS-SPIONs was confirmed by dynamic light scatter and infrared spectrometer,and the contents of iron and polysaccharide were characterized by spectrophotometry.The effect of APS-SPIONs on inducing mouse macrophages RAW264.7 polarization was investigated by flow cytometry.The RAW264.7 macrophages-HepG2 human hepatoma cancer cells Transwell co-culture system was established to investigate APS-SPIONs improve anti-tumor function of macrophages in vitro,and the proliferation activity of APS-SPIONs on RAW264.7 detected by cell counting kit-8(CCK-8)method.The results showed that the average particle size and zeta potential of APS-SPIONs were(82.93±1.47)nm and(-24.00±0.47)mV.Polysaccharide and Fe content were 8.69%and 7.04%,respectively.APS-SPIONs effectively induced the polarization of RAW264.7 into M1 type in vitro,improving the anti-tumor ability of macrophages in a co-culture system,without effecting the proliferation of macrophages.Our study provides a drug development strategy and preliminary research results to educate macrophages and reshape the tumor immune microenvironment to achieve tumor-killing effects.

关 键 词：磁性氧化铁纳米粒 黄芪多糖 巨噬细胞极化 肿瘤相关巨噬细胞 纳米药物 

分 类 号：R945[医药卫生—微生物与生化药学]