透明质酸修饰的pH响应型阿霉素纳米递送系统靶向多发性骨髓瘤的体外研究  

Hyaluronic acid-modified pH-responsive doxorubicin nano-delivery system targeting multiple myeloma in vitro

作　　者：阎佳画 张传敏 肖伯莲 王丽娟 YAN Jiahua;ZHANG Chuanmin;XIAO Bolian;WANG Lijuan(Linyi People's Hospital Cultivation Base,Jinzhou Medical University,Shandong Linyi 276000,China;Central Laboratory,Linyi People's Hospital,Shandong Linyi 276000,China;Linyi Key Laboratory of Tumor Biology,Shandong Linyi 276000,China;Key Laboratory of Neurophysiology,Health Commission of Shandong Province,Shandong Linyi 276000,China;XZMU Key Laboratory Cultivation Base for Translational Oncology,Shandong Linyi 276000,China;Department of Hematology,Linyi People's Hospital,Shandong Linyi 276000,China)

机构地区：[1]锦州医科大学临沂市人民医院培养基地,山东临沂276000 [2]临沂市人民医院中心实验室,山东临沂276000 [3]临沂市肿瘤生物学重点实验室,山东临沂276000 [4]山东省卫生健康委员会医药卫生神经生理学重点实验室,山东临沂276000 [5]徐州医科大学肿瘤转化医学重点实验室培育建设点,山东临沂276000 [6]临沂市人民医院血液内科,山东临沂276000

出　　处：《现代肿瘤医学》2023年第11期1973-1979,共7页Journal of Modern Oncology

基　　金：国家自然科学基金项目(编号:81402353);中国博士后科学基金(编号:2015M580594);山东省重点研发计划(编号:2018GSF118035);徐州医科大学附属医院发展基金(编号:XYFM2020016)。

摘　　要：目的:构建一种特异性靶向多发性骨髓瘤的pH响应型纳米递送系统,实现对多发性骨髓瘤细胞靶向释放化疗药物阿霉素。方法:采用酸敏感的DSPE-PEOz和阳离子类脂DOTAP包封模型药物阿霉素(doxorubicin,DOX),得到阿霉素纳米递送系统(DOX-NDS),并经HA-PEG_(2000)-DSPE(HA)靶向长循环修饰,最终制得阿霉素靶向纳米递送系统(DOX-HA-NDS);纳米粒度电位仪分析DOX-HA-NDS的粒径和Zeta电位;多功能酶标仪检测DOX-HA-NDS的包封率(encapsulation efficiency,EE)和载药量(drug loading,DL);透析法研究DOX-HA-NDS在pH 5.0和pH 7.4条件下的体外释药行为;流式细胞仪检测其细胞摄入;CCK-8法评价空白靶向纳米递送系统(Blank-HA-NDS)的毒性和DOX-HA-NDS的细胞增殖抑制作用。结果:构建的DOX-HA-NDS粒径为(193.1±5.0)nm,Zeta电位为(-41.1±2.0)mV,离心法和透析法测得包封率分别高达93%和90%,载药量为32.76%和32%,4℃存储能稳定6个月以上。在pH 7.4条件下,DOX-HA-NDS的释药缓慢,且6 h累积释放率仅为30%,而在pH 5.0条件下,DOX-HA-NDS的释药明显加快,6 h累积释放率高达97%,表明构建的阿霉素靶向纳米递送系统具有pH响应控释性能。细胞摄入实验结果表明,经HA修饰后的阿霉素纳米递送系统可以更有效靶向肿瘤细胞;体外抗肿瘤活性结果表明,Blank-HA-NDS基本无细胞毒性,DOX-HA-NDS对人多发性骨髓瘤细胞(ARH-77)的增殖抑制作用最强。结论:构建的阿霉素靶向纳米递送系统不仅包封率高,而且兼具主动靶向和pH响应控释性能,提高对DOX的输送效率,从而增强了其对ARH-77的细胞增殖抑制作用。Objective:To construct a pH-responsive nano-delivery for specific targeting multiple myeloma cells to realize the controlled release of chemotherapeutic drug DOX.Methods:Doxorubicin targeted nano-delivery system(DOX-HA-NDS)was designed and constructed by using DSPE-PEOz,DOTAP and HA-PEG_(2000)-DSPE(HA)for delivery of the model drug doxorubicin(DOX).The particle size and Zeta potential of DOX-HA-NDS were analyzed by using nanoparticle size analyzer.The encapsulation efficiency and drug loading of DOX-HA-NDS were detected by microplate reader.The drug release behavior of DOX-HA-NDS in vitro(pH 5.0 and pH 7.4)was studied by dialysis method.The cell uptake of DOX-HA-NDS was detected by flow cytometry.The toxicity of blank nano-delivery system(Blank-HA-NDS)and the inhibitory effect of DOX-HA-NDS on cell proliferation were evaluated by CCK-8 assay.Results:The obtained DOX-HA-NDS with particle size of(193.1±5.0)nm,Zeta potential of(-41.1±2.0)mV,the high encapsulation efficiency of 93% and 90%,and the drug loading efficiency of 32.76% and 32% with centrifugation and dialysis method,could maintain its stability at least 6 months at 4℃.At pH 7.4,the cumulative DOX release from DOX-HA-NDS was slow and the cumulative release rate for DOX-HA-NDS was only 30% at 6 h,while at pH 5.0,the DOX release fraction increased significantly and the cumulative release rate for DOX-HA-NDS was up to 97%within 6 h,indicating DOX-HA-NDS has pH-responsive controlled release behavior.The results of cell uptake test showed that compared with DOX-NDS,nano-delivery system modified with hyaluronic acid(DOX-HA-NDS)could target tumor cells more effectively.The results of in vitro antitumor activity showed that the blank nano-delivery system(Blank-HA-NDS)had low cytotoxicity.At the same time,compared with Free DOX and DOX-NDS,DOX-HA-NDS had the strongest cell growth inhibition on human multiple myeloma cells(ARH-77).Conclusion:The constructed DOX-HA-NDS with high encapsulation efficiency,active targeting ability and pH-responsive controlled releas

关 键 词：纳米递送 阿霉素 主动靶向 pH响应控释 CD44 

分 类 号：R733.3[医药卫生—肿瘤]