β-榄香烯聚合物胶束的构建及体外抗肿瘤作用研究  

Construction ofβ-elemene polymer micelles and its antitumor effect in vitro

作　　者：孙吴倩 邵麒 贾玉倩 郭秋红 刘兴超 周莉 左赛杰 SUN Wuqian;SHAO Qi;JIA Yuqian;GUO Qiuhong;LIU Xingchao;ZHOU Li;ZUO Saijie(College of Pharmacy,Hebei University of Chinese Medicine,Shijiazhuang 050091,China;Hebei Technology Innovation Center of TCM Formula Preparations,Shijiazhuang 050091,China;Hebei Higher Education Institute Applied Technology Research Center on TCM Development and Industrialization,Shijiazhuang 050091,China)

机构地区：[1]河北中医药大学药学院,河北石家庄050091 [2]河北省中药组方制剂技术创新中心,河北石家庄050091 [3]河北省高校中药开发与产业化应用技术研发中心,河北石家庄050091

出　　处：《沈阳药科大学学报》2023年第11期1409-1416,1465,共9页Journal of Shenyang Pharmaceutical University

基　　金：河北省自然科学基金项目(H2020423006);河北省高等学校青年拔尖人才计划项目(BJK2022034);河北中医学院博士科研基金项目(BSZ2019004);河北中医学院优秀青年教师基础研究计划项目(YQ2019010)。

摘　　要：目的以甲氧基聚乙二醇-聚己内酯(MPEG-PCL,MP)嵌段共聚物为载体,β-榄香烯(β-Elemene,β-Ele)为主药成分制备榄香烯纳米胶束(β-Ele/MP),并对β-Ele/MP纳米胶束进行表征以及体外释放考察;以A549细胞为体外抗肿瘤作用研究对象,考察β-Ele溶液,β-Ele/MP纳米胶束对A549细胞增殖活性的抑制作用,在荧光显微镜下观察细胞对纳米胶束的摄取行为。方法采用薄膜分散法制备β-Ele/MP纳米胶束,并对其粒径、Zeta电位、载药量、包封率等进行表征;以不同pH值的磷酸盐缓冲液(pH=5.5、pH=6.8、pH=7.4)为释放介质对β-Ele/MP纳米胶束进行体外释放的考察。采用CCK-8法测定β-Ele溶液、β-Ele/MP纳米胶束对A549细胞增殖活性的抑制作用;以香豆素6为荧光探针,Hochest33342对细胞核进行染色定位,荧光显微镜下观察细胞对胶束的摄取行为。结果制备得到β-Ele/MP纳米胶束平均粒径是(35.8±0.212)nm,多分散系数(PDI)为0.093±0.001,Zeta电位是(-17.3±0.721)mV,包封率和载药量分别为(93.9±1.124)%,(5.9±1.212)%;体外释放的结果显示β-Ele/MP纳米胶束72 h内最高释放量达64.56%,遵循Weibull方程;体外抗肿瘤实验结果表明,β-Ele/MP纳米胶束对A549细胞增殖活性的抑制作用要强于β-Ele溶液;β-Ele/MP纳米胶束相比于β-Ele溶液的IC 50值有明显的降低;荧光显微镜下观察A549细胞对纳米胶束的摄取行为,结果表明随着时间的延长,细胞对药物摄取量增加。结论本实验结果表明β-Ele纳米胶束在抗肿瘤方面具有应用前景,可为改善抗肿瘤药物的开发与应用提供更多的理论和实验依据。Objectiveβ-Elemene nano-micelles(β-Ele/MP)were prepared using methoxy polyethylene glycol polycaprolactone(MPEG-PCL,MP)block copolymer as a carrier andβ-Elemene(β-Ele)as a main component.β-Ele/MP nano-micelles were characterized and their release in vitro were evaluated.Taking A549 cells as the object of in vitro anti-tumor study,the inhibitory effects ofβ-Ele solution andβ-Ele/MP nano-micelles on the proliferative activity of A549 cells were investigated,and the uptake of nano-micelles by cells was observed under fluorescence microscope.Methodsβ-Ele/MP nano-micelles were prepared by thin film dispersion method,and their particle size,Zeta potential,drug loading and encapsulation efficiency were characterized.The release in vitro ofβ-Ele/MP nano-micelles was evaluated using phosphate buffers(pH=5.5,pH=6.8,pH=7.4)at different pH values.The inhibitory effects ofβ-Ele solution andβ-Ele/MP nano-micelles on A549 cell proliferation were determined by CCK-8 method.The uptake behavior of nano-micelles was observed under fluorescence microscope by using coumarin 6 as a fluorescence probe and Hochest33342.Results The average particle size ofβ-Ele/MP micelles was(35.8±0.212)nm,poly-dispersion coefficient(PDI)was 0.093±0.001,Zeta potential was(-17.3±0.721)mV,encapsulation efficiency and drug loading were(93.9±1.124)%,(5.9±1.212)%,respectively.The release ofβ-Ele/MP nano-micelles in vitro was 64.56%within 72 h,which followed the Weibull equation.In vitro test results showed that the inhibitory effect ofβ-Ele/MP nano-micelles on the proliferation activity of A549 cells was stronger than that ofβ-Ele solution.Compared withβ-Ele solution,IC 50 value ofβ-Ele/MP nano-micelles was significantly decreased.The uptake behavior of nano-micelles by A549 cells was observed under a fluorescence microscope,and the results showed that the drug uptake by cells increased with time.Conclusion Theβ-Ele nano-micelles have a promising application in antitumor drugs,and can provide more theoretical and experimental basis for

关 键 词：甲氧基聚乙二醇-聚己内酯 纳米胶束 Β-榄香烯 A549细胞 体外释放 

分 类 号：R94[医药卫生—药剂学]