叶酸偶联紫杉醇纳米粒的制备及其生物效应研究

发布时间：2018-11-24 08:05

【摘要】：目的以叶酸修饰的生物可降解材料乳酸-羟基乙酸共聚物(PLGA-PEG-FA)为载体,构建叶酸偶联紫杉醇纳米粒〔PLGA-PEG-FA(PTX)-NPs〕并进行评价。研究其对MCF-7细胞的效应并初步探讨其靶向作用机制。方法乳化分散法制备PLGA-PEG-FA(PTX)-NPs,并对纳米粒的形态、粒径、Zeta电位、包封率及载药量进行评价;同法制备无叶酸偶联的紫杉醇纳米粒〔PLGA-PEG(PTX)-NPs〕,分别用荧光物质FITC标记,应用荧光显微镜观察其对MCF-7细胞和A549细胞的靶向作用,以及游离叶酸对PLGA-PEG-FA(PTX)-NPs的干扰作用;MTT法观察PLGA-PEG-FA(PTX)-NPs、PLGA-PEG(PTX)-NPs对MCF-7细胞的生长抑制作用;流式细胞术检测细胞凋亡率。结果制备的PLGA-PEG-FA(PTX)-NPs为均匀球形粒子,粒径为(88.2±6.7)nm,Zeta电位为(56.5±4.2)mV,包封率为(92.96±0.16)%,载药量为(4.84±0.10)%。与PLGA-PEG(PTX)-NPs相比,PLGA-PEG-FA(PTX)-NPs能更有效的被MCF-7细胞吞噬(P0.05);PLGA-PEG-FA(PTX)-NPs、PLGA-PEG(PTX)-NPs对A549细胞无明显靶向作用;培养基中加入游离叶酸干扰了PLGA-PEG-FA(PTX)-NPs的靶向性。MTT法测得PLGA-PEG-FA(PTX)-NPs抑制MCF-7细胞生长的IC50值(10.72μg·ml~(-1))明显低于PLGAPEG(PTX)-NPs组(17.41μg·ml~(-1),P0.05),紫杉醇浓度在1~15μg·ml~(-1)时,PLGA-PEG-FA(PTX)-NPs组的生长抑制率在24h明显高于PLGA-PEG(PTX)-NPs组(P0.05),紫杉醇浓度达到20μg·ml~(-1)时,两组无显著性差异。PLGA-PEG(PTX)-NPs组的48h细胞凋亡率(87.04%)明显高于PLGA-PEG(PTX)-NPs组(64.14%)。结论制备的PLGA-PEG-FA(PTX)-NPs能够靶向抑制MCF-7细胞增殖并促进其凋亡,其靶向性可能是由叶酸受体介导的。
[Abstract]:Objective to construct folic acid-coupled paclitaxel nanoparticles (PLGA-PEG-FA (PTX)-NPs) using folic acid modified biodegradable material lactic acid-glycolic acid copolymer (PLGA-PEG-FA) as the carrier. To study its effect on MCF-7 cells and the mechanism of its targeting. Methods PLGA-PEG-FA (PTX)-NPs, was prepared by emulsification and dispersion method. The morphology, particle size, Zeta potential, encapsulation efficiency and drug loading capacity of nanoparticles were evaluated. Paclitaxel nanoparticles without folic acid coupling (PLGA-PEG (PTX) NPs),) were prepared by the same method and labeled with fluorescent substance FITC, and their targeting effects on MCF-7 and A549 cells were observed by fluorescence microscope. And the interference of free folic acid to PLGA-PEG-FA (PTX)-NPs; The inhibitory effect of PLGA-PEG-FA (PTX)-NPs,PLGA-PEG (PTX)-NPs on the growth of MCF-7 cells was observed by MTT assay, and the apoptosis rate was detected by flow cytometry. Results the PLGA-PEG-FA (PTX) NPs particles were homogeneous spherical particles, the particle size was (88.2 卤6.7) nm,Zeta potential was (56.5 卤4.2) mV, entrapment efficiency was (92.96 卤0.16)%, the drug loading was (4.84 卤0.10)%. Compared with PLGA-PEG (PTX) NPs, PLGA-PEG-FA (PTX) NPs was more effectively phagocytized by MCF-7 cells (P0.05), PLGA-PEG-FA (PTX) NPs,PLGA-PEG (PTX) NPs had no significant targeting effect on A549 cells. The concentration of free folic acid in the medium interfered with the targeting of PLGA-PEG-FA (PTX)-NPs. The IC50 value of PLGA-PEG-FA (PTX)-NPs in inhibiting the growth of MCF-7 cells (10. 72 渭 g ml~ (- 1) was significantly lower than that of PLGAPEG by MTT assay (PTX) NPs group (17.41 渭 g ml~ (-1), When the concentration of paclitaxel was 15 渭 g ml~ (-1), the growth inhibition rate of PLGA-PEG-FA (PTX)-NPs group was significantly higher than that of PLGA-PEG (PTX)-NPs group at 24 h (P0.05), and taxol concentration reached 20 渭 g ml~ (-1). There was no significant difference between the two groups. The apoptosis rate in PLGA-PEG (PTX)-NPs group (87.04%) was significantly higher than that in PLGA-PEG (PTX)-NPs group (64.14%). Conclusion the prepared PLGA-PEG-FA (PTX)-NPs can inhibit the proliferation and promote the apoptosis of MCF-7 cells, which may be mediated by folic acid receptor.
【作者单位】：空军总医院药学部;
【基金】：全军医学科技青年培育项目基金,No.13QNP079
【分类号】：R943

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