秦利芳,林东海,李刚,谢鑫鑫,王俊腾,闻真
QIN Li-fang1,LIN Dong-hai1,LI Gang2,XIE Xin-xin1,WANG Jun-teng1,WEN Zhen1(1.School of Pharmacy

• 1:烟台大学药学院
• 2:烟台大学生命科学学院

摘要(Abstract)：

目的制备单甲氧基聚乙二醇-聚乳酸-羟基乙酸纳米粒(monomethoxy polyethylene glycol-poly-actic coglycolic acid-nanoparticles,mPEG-PLGA-NPs)并研究其理化性质及体外抗黏性能,考察人宫颈癌细胞HeLa对mPEG-PLGA-NPs的摄取能力。方法采用溶剂扩散法制备mPEG-PLGA-NPs,测定其平均粒径和zeta电位。采用黏蛋白结合法,考察mPEG-PLGA-NPs的体外抗黏性能。以香豆素-6(coumarin 6)为荧光标记物,通过共聚焦显微镜及HPLC法进行HeLa细胞的体外摄取研究。结果 mPEG-PLGA-NPs的平均粒径为(106.2±4.3)nm,Zeta电位为-(12.40±0.11)mV。PLGA-NPs的抗黏蛋白黏附率为35.5%,而mPEG-PLGA-NPs的抗黏蛋白黏附率为92.5%,比PLGA-NPs高3倍左右。相同孵育时间内,HeLa细胞对mPEG-PLGA-NPs的摄取量是PLGA-NPs的近2倍,HeLa细胞对PLGA-NPs和mPEG-PLGA-NPs的摄取量与孵育时间成明显的依赖关系。结论 mPEG-PLGA-NPs具有较强的抗黏性;HeLa细胞对mPEG-PLGA-NPs的摄取明显高于PLGA-NPs(P<0.01),表现出更好的细胞亲和性。
Objective To prepare mPEG-PLGA nanoparticles,discuss its properties and in vitro anti-adhesive activity and investigate the cellular uptake of them in human cervical cancer cells(HeLa).Methods mPEG-PLGA-NPs were prepared by a solvent diffusion method.The particle size distribution and Zeta potential of nanoparticles were measured by light scattering.The anti-adhesive activity of nanoparticles was evaluated by pig gastric mucin(PM) binding experiments.Coumarin 6 was incorporated into nanoparticles as fluorescent marker.The cellular uptake of PLGA-NPs and PEG-PLGA-NPs by HeLa cell lines were determined by laser scanning confocal microscope and HPLC.Results The mean size of mPEG-PLGA-NPs was(106.2±4.3)nm,Zeta potential was-(12.40±0.11)mV.The PM anti-adhesive rate of PLGA-NPs and PEG-PLGA-NPs was 35.5% and 92.5%,respectively.The anti-adhesive activity of mPEG-PLGA-NPs was three times higher than PLGA-NPs.All nanoparticles showed low cytotoxicity.At the same incubation time,the uptake of mPEG-PLGA-NPs was two times higher than that of unmodified PLGA-NPs.The cellular uptake of PLGA-NPs and mPEG-PLGA-NPs was dependent on the incubation time.Conclusions mPEG-PLGA-NPs has much higher anti-adhesive activity.The uptake of mPEG-PLGA-NPs by HeLa cells is much higher compared to the PLGA-NPs(P<0.01).The mPEG-PLGA-NPs shows better cell affinity than PLGA-NPs.

关键词(KeyWords)： 单甲氧基聚乙二醇-聚乳酸-羟基乙酸;纳米粒;抗黏性;HeLa细胞;细胞摄取
mPEG-PLGA;nanoparticle;anti-adhesive activity;HeLa cell line;cellular uptake

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(30973949);; 山东省自然科学基金资助项目(ZR2009CM012)

作者(Author): 秦利芳,林东海,李刚,谢鑫鑫,王俊腾,闻真
QIN Li-fang1,LIN Dong-hai1,LI Gang2,XIE Xin-xin1,WANG Jun-teng1,WEN Zhen1(1.School of Pharmacy

DOI: 10.14066/j.cnki.cn21-1349/r.2012.12.014

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