荷载Zebularine的聚合物胶束复合体纳米颗粒对体外培养的晶状体上皮细胞增殖和黏附的影响

Effect of Zebularine loaded MePEG-PCL nanoparticles on viability,attachment of in vitro cultured lens epithelial cells

目的:研究荷载Zebularine(Zeb)的聚合物胶束复合体(MePEG-PCL)纳米颗粒(NPs)对体外培养的晶状体上皮细胞(LECs)活性、黏附性和凋亡的影响。方法:体外培养胎儿晶状体组织HLE B-3永生细胞,分为6个组,分别给予游离Zeb 50μmol/L(ZebF1组)、100μmol/L(ZebF2组)、荷载Zeb的MePEG-PCL NPs 50μmol/L(ZebNP1组)、100μmol/L(ZebNP2组)、MePEG-PCL空载颗粒(NPs组)、空白培养基(C组),分别采用MTT比色法、改良MTT比色法观察细胞活性、黏附性,采用DNA ladder法研究细胞凋亡。结果:MTT比色法显示:游离Zeb和荷载Zeb的MePEG-PCL NPs对细胞活性有抑制作用,均呈时间-剂量依赖性增强(P<0.05),与游离组相比,相同剂量荷载Zeb的MePEG-PCL NPs组在给药12h时细胞活性无差异,而给药24,48,96h后载体组细胞活性降低(P<0.05)。改良MTT比色法显示:所有Zeb给药组细胞黏附率均降低,与游离组相比相同剂量荷载Zeb的MePEG-PCL NPs组黏附率降低(P<0.05)。DNA ladder显示:给药后96h C组、NPs组,ZebF1组未出现DNA碎片表达,ZebF2,ZebNP1,ZebNP2组出现DNA碎片表达,其表达强度依次为ZebNP2>ZebNP1>ZebF2(P<0.05)。结论:荷载Zeb的MePEG-PCL NPs可有效地抑制体外培养的LECs活性、黏附性,造成细胞的凋亡,其效果优于同剂量的游离药物。

AIM： To investigate the effect of zebularine（ Zeb）loaded Poly（ ethylene glycol）- block- poly（ ε-caprolactone） methyl ether（ MePEG- PCL） nanoparticles（ NPs） on the viability,attachment,and apoptosis of in vitro cultured lens epithelial cells（ LECs）.METHODS： In vitro cultured infant human lens tissue HLE B- 3 immortalized cells were distributed randomly divided into six groups. Each group was administered with free Zeb 50μ mol /L（ ZebF1 group）,100μ mol /L（ ZebF2group）, Zeb- loaded MePEG- PCL NPs 50μ mol /L（ ZebNP1 group）,Zeb-loaded MePEG-PCL NPs100μmol/L（ ZebNP2 group）,MePEG- PCL empty NPs（ NPs group）or blank medium（ group C） respectively. A tetrazolium dye assay（ MTT） test and modified MTT test were performed to determine cell viability and cell attachment.DNA ladder was used to detect the cell apoptosis.RESULTS： Determined by MTT colorimetric method：Cell proliferation rate of LECs were suppressed by all Zeb administration groups in a concentration- time dependent manner（ P 〈 0. 05）. Compared with the free Zeb groups,the viability of LECs were suppressed more effectively by the same dose of Zeb loaded MePEG- PCL NPs after 24,48,96h（ P 〈 0. 05）. Determined by improved MTT colorimetric method： The attachment of LECs were decreased in all Zeb administration groups,Zeb loaded MePEG- PCL NPs had better effect on suppressing the attachment of LECs than the free Zeb groups with same dose（ P 〈 0. 05）. The DNA ladder confirmed that after administration of 96 h,group C,NPs group and ZebF1 group showed no DNA fragment, however the DNA fragment were performed in ZebF2, ZebNP1, ZebNP2 groups and displays the trend of ZebNP2 〉 ZebNP1 〉ZebF2（ P 〈 0. 05）.CONCLUSION： Zeb loaded MePEG-PCL NPs had better effect on suppressing the viability and attachment of in vitro cultured LECs than the free Zeb groups,as well as enhancing the apoptosis.