摘要
目的以组氨酸(histidine,His)修饰的透明质酸(hyaluronic acid,HA)为载体,阿霉素(doxorubicin,DOX)为模型药物,制备纳米粒DOX/His-HA,并分析其理化性质和体内外抗肿瘤活性。方法通过化学交联法将组氨酸与HA进行结合,采用超声法制备纳米粒DOX/His-HA。用DAWN HELEOSⅡ动态激光光散射仪测定His-HA纳米粒的粒径和Zeta电位,透射电镜观察其形态;紫外-可见分光光度计测定DOX/His-HA纳米粒中DOX的吸光度值,计算其包封率、载药量和DOX的体外释放度。采用MTT法检测DOX/His-HA对Hep G2细胞的生长抑制作用;摄入试验观察Hep G2细胞对DOX/His-HA的摄入情况;抑瘤试验分析DOX/His-HA的体内抗肿瘤活性。结果 His-HA纳米粒呈球形,粒径为230~480 nm,分散度良好,Zeta电位为-19.3^-13.5 m V。DOX/His-HA的载药量和包封率分别达到(8.14±0.09)%和(91.37±0.69)%。在前4 h内,DOX/His-HA纳米粒均存在明显突释现象;4 h后,DOX/HisHA纳米粒逐渐表现出一定的缓释性。随着组氨酸取代度的增加,DOX/His-HA的Hep G2细胞毒性增强。组氨酸取代度越高,DOX/His-HA越易被细胞摄入。DOX/His-HA具有更强的抑瘤效果,且对正常组织无明显的毒副作用。结论制备的DOX/His-HA纳米粒是一种缓释性和肿瘤靶向性良好的给药系统。
Objective To prepare hyaluronic acid nanoparticles（DOX / His-HA）by using histidine modified hyaluronic acid as the carrier and adriamycin（DOX） as a model drug, and analyze their physicochemical property and anti-tumor activity in vitro and in vivo. Methods Histidine and hyaluronic acid were combined by chemical cross-linking method,based on which DOX / His-HA nanoparticles were prepared by ultrasonic method, and determined for particle size and Zeta potential by DAWN HELEOS Ⅱ dynamic laser particle size analyzer, and observed for morphology by transmission electron microscopy. The absorbance of DOX in DOX / His-HA nanoparticles was determined by ultraviolet-visible spectrophotometer, while the encapsulation efficiency, drug load and in vitro release were calculated. MTT assay was used to determine the inhibitory effect of DOX / His-HA on growth of Hep G2 cells, while intake test to observe the intake of DOX / His-HA by Hep G2 cells, and tumor inhibition test to analyze the antitumor activity in vivo of DOX / His-HA.Results His-HA nanoparticles were in spherical form with a good dispersion, at diameters of 230 ～ 480 nm, of which the Zeta potential was-19. 3 ～-13. 5 m V. The drug load and encapsulation efficiency of DOX / His-HA nanoparticles were（8. 14 ± 0. 09）% and（91. 37 ± 0. 69）% respectively. Burst release was observed in DOX / His-HA nanoparticles within 4 h, while controlled release 4 h later. The cytotoxicity of DOX / His-HA to Hep G2 cells increased with the increasing degree of His substitution. The higher the His substitution degree was, the easier the intake of DOX / His-HA was. DOX / His-HA showed strong anti-tumor effect, without obvious side effects on normal tissue. Conclusion The prepared DOX / His-HA nanoparticles was a control-released and tumor-targeting drug delivery system.
出处
《中国生物制品学杂志》
CAS
CSCD
2016年第6期600-605,共6页
Chinese Journal of Biologicals
基金
国家自然科学基金(81274093)
山东省自然科学基金(ZR2013CL026)
山东医药卫生计划(2013WS0284)
山东中医药卫生课题(2015232)
潍坊市中医药发展项目(2005001)
关键词
阿霉素
透明质酸
纳米粒
抗肿瘤活性
Doxorubicin（DOX）
Hyaluronic acid
Nanoparticles
Anti-tumor activity
