pH响应性PTEN/PLGA-(HE)_(10)-MAP纳米粒的构建及体外评价  

Construction and in vitro evaluation of pH-responsive and tumor-targeted PTEN/PLGA-(HE)_(10)-MAP nanoparticles

作　　者：雍琴 岳瀚勋 石敏 黄仕琴 赵轩 余娴[1] YONG Qin;YUE Hanxun;SHI Min;HUANG Shiqin;ZHAO Xuan;YU Xian(Phase I Clinical Trial Center,The Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010;Department of Pharmacy,Central Hospital of Luohe,Luohe 462000;Department of Pharmacy,Central Hospital of Jiangjin District,Chongqing 402260,China)

机构地区：[1]重庆医科大学附属第二医院Ⅰ期临床试验研究室,重庆400010 [2]漯河市中心医院药学部,漯河462000 [3]重庆市江津区中心医院药学部,重庆402260

出　　处：《中国药科大学学报》2021年第3期301-310,共10页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.82072327)。

摘　　要：构建一种pH响应性细胞穿膜肽(cell-penetrating peptides,CPPs)修饰的载抑癌基因第10号染色体同源缺失性磷酸酯酶-张力蛋白(PTEN)质粒DNA的纳米粒PTEN/PLGA-(HE)_(10)-MAP,探讨其基因递送和体外靶向抗肿瘤作用。采用双乳化-溶剂挥发法制备载PTEN质粒DNA的聚乳酸-羟基乙酸共聚物(PLGA)纳米粒PTEN/PLGA;利用酰胺缩合反应将pH响应性组氨酸-谷氨酸(HE)重复寡肽与模型两亲性多肽(MAP)的重组体(HE)10-MAP偶联至PLGA纳米粒表面,得到纳米粒PTEN/PLGA-(HE)_(10)-MAP。以粒径、Zeta电位以及包封率与载药量为指标对其进行表征分析;通过考察其细胞毒性、细胞摄取,以及靶向转染真核表达质粒和抗肿瘤细胞增殖的能力,分析其作为目的基因靶向递送系统的可行性。结果显示,制备的纳米粒粒径为(2`66.5±2.86)nm,包封率为(80.6±6.11)%,在pH 7.4、7.0和6.5条件下Zeta电位分别为-(6.7±0.26)mV、+(0.7±0.22)mV和+(37.5±0.85)mV;未载质粒DNA的空载体纳米粒PLGA-(HE)_(10)-MAP在肿瘤和正常细胞中的细胞毒性试验显示细胞存活率均在80%以上,制备的纳米粒可以被细胞摄取表达,且具有pH靶向抑制肿瘤细胞增殖的作用,在肿瘤的基因治疗中具有一定的应用前景。To construct PTEN/PLGA-(HE)_(10)-MAP nanoparticles,which encapsulated PTEN plasmid DNA andcombined with the p H-responsive cell-penetrating peptides(CPPs),and to investigate their effects of genedelivery and anti-tumor targets in vitro.Poly(lactic-co-glycolic acid)(PLGA)nanoparticles loaded with PTENplasmid DNA were prepared by double emulsification-solvent evaporation method.PTEN/PLGA-(HE)_(10)-MAPnanoparticles were prepared by coupling the histidine-glutamic acid-model amphipathic peptide nanocomplex[(HE)_(10)-MAP]to the surface through amide condensation reaction.Particle size,Zeta potential,encapsulationrate and drug loading were tested to characterize the nanoparticles.By analyzing the cytotoxicity,cellularuptake,targeted transfection of eukaryotic expression plasmids and anti-tumor cell proliferation,the feasibility asa targeted gene delivery system were evaluated.The particle size of PTEN/PLGA-(HE)_(10)-MAP nanoparticles was(266.5±2.86)nm,with the encapsulation efficiency(80.6±6.11)%.Zeta potentials were-(6.7±0.26)m V,+(0.7±0.22)m V and+(37.5±0.85)m V at p H 7.4,7.0 and 6.5,respectively.In the cytotoxicity test,thecell survival rates of tumor and normal cells were above 80%.Non-loading PLGA-(HE)_(10)-MAP nanoparticlesshowed no obvious cytotoxicity.The results of cellular uptake experiments showed that PTEN/PLGA-(HE)_(10)-MAP nanoparticles were more readily taken up by cells.The results of CCK-8 showed that the nanoparticlescould p H-specifically inhibit proliferation of tumor cell in vitro.And PTEN/PLGA-(HE)_(10)-MAP nanoparticles maybe applied in tumor gene therapy.

关 键 词：纳米粒 PH响应 PLGA 肿瘤靶向 细胞穿膜肽 基因递送 

分 类 号：R944[医药卫生—药剂学]