【作者】 李博； 李贞； 王成润； 金一；

【Author】 LI Bo,LI Zhen,WANG Chengrun,JIN Yi College of pharmaceutical Sciences,Zhejiang University,Hangzhou 310058,China

【机构】 浙江大学药学院；

【摘要】 目的制备c(RGD)fk介导的阿霉素-双氢青蒿素纳米粒脂质体(lipoparticles),并建立小鼠S180肉瘤及B16黑色素瘤模型,对其抗肿瘤效果进行评价。方法以聚乳酸/羟乙酸-阿霉素接枝共聚物(PLGA-DOX),二棕榈酰磷脂酰胆碱(DPPC)-双氢青蒿素磷脂复合物为载体材料,制备阿霉素-双氢青蒿素纳米粒脂质体,合成了c(RGD)fk-聚乙二醇-胆固醇接枝共聚物(c(RGD)fk-PEG-Chol)并对纳米粒脂质体进行修饰;应用动态光散射粒度仪,透射电子显微镜、扫描电子显微镜,对载药纳米粒的Zeta电位、粒径与形态进行评价;c(RGD)fk修饰后的纳米粒脂质体对内皮细胞的结合率明显提高,建立ICR小鼠S180肉瘤模型和B16黑色素瘤模型,观察制剂的抑瘤效果,以及对肿瘤VEGF、HIF-1及VWF蛋白表达的影响。结果 PBS7.4缓冲溶液中,通过磷脂层的包裹,纳米粒的Zeta电位由正电性变为中性,粒径增大,形态呈球形。透射电子显微镜和扫描电子显微镜结果显示包裹后的纳米粒表面结构发生了明显的变化。ICR小鼠S180皮下移植瘤模型和肺部B16瘤细胞模型表明,c(RGD)fk可提高纳米粒脂质体的肿瘤组织血管靶向性,发挥双氢青蒿素与阿霉素的协同治疗效果,改善荷瘤小鼠的生命体征,提高抑瘤率最高达80.5%;免疫组织化学检测表明,c(RGD)fk介导的脂质体纳米粒组可有效降低肿瘤组织COX-2、VEGF、HIF-1及VWF的表达。结论细胞实验表明c(RGD)fk介导的纳米粒脂质体可有效的靶向到内皮细胞,提高制剂的靶向性,最大限度的发挥联合用药的效果,抑制肿瘤的生长和转移。更多还原

【Abstract】 OBJECTIVE The lipoparticles loaded doxorubicin and dihydroartemisinin modified by c(RGD)fk were prepared here,and the physicochemical and antitumor effect of c(RGD)fk-LNP was studied with experimental tumor S180 and B16 mice.METHODS PLGA and DPPC were used to prepare lipoparticles,and the characteristics of LNP was analyzed using dynamic light scattering,TEM and SEM.The result of fluorescent microscopy showed that the uptake of c(RGD)fk-liposome by HUVEC could be increased;Experimental tumor S180 and B16 mice was used to observe the anti-tumor effect of c(RGD)fk-LNP,and the protein expressions of VEGF,HIF-1,and VWF were evaluated.RESULTS The nanoparticles were almost spherical in shape.Their size increased from 100nm to about 200nm,and the zeta potential was decreased from 58mv to -7.6mv which indicated the cover of DPPC.The tumor inhibitory rate of c(RGD)fk-LNP group is 80.5%,which was better than other single dose groups.The combined drug therapy of DHA and DOX can significantly inhibit tumor growth. The cyclopeptide c(RGD)fk can target to the microvessel of tumor and show less cytotoxicity,and the expression of VEGF,HIF-1and MVD were down regulated.Histopathological examination showed the inhibition of angiogenesis and metastasis in tumor.更多还原