反义寡聚核苷酸对兔角膜基质细胞的通透性

Permeability of antisense oligonucleotides through rabbit keratocytes

目的 :观察反义寡聚核苷酸及其脂质体对角膜基质细胞的通透性。方法 :将荧光标记的 10 0、2 0 μg/ml反义寡聚核苷酸与 5 0、10 μg/ml的反义寡聚核苷酸脂质体加入至兔角膜细胞培养板中 ,分别在 0 5和 4小时 ,荧光显微镜下观察。结果 :0 5小时 ,10 0 μg/ml反义寡聚核苷酸和 5 0、10 μg/ml反义寡聚核苷酸脂质体组细胞内有荧光 ,而 2 0 μg/ml的反义寡聚核苷酸组无荧光 ;4小时 ,所有实验组细胞内都有荧光 ,但 2 0 μg/ml反义寡聚核苷酸组荧光细胞数量少。结论 :游离反义寡聚核苷酸可进入角膜细胞中 。

Objective:To investigate the permeability of antisense oligonucleotides and their liposomes through rabbit keratocytes.Methods:Oligonucleotides(100 and 20μg/ml) and their liposomes (50 and 10μg/ml) of FITC were put into the plates of keratocytes,which were observed with fluorescence microscope 1/2 hour and 4 hours later.Results:The fluorescence in the keratocytes was found in the groups of oligonucleotides (100μg/ml) and their liposomes (50 and 10μg/ml) after 0.5 hour,and not in the group of oligonucleotides (20μg/ml).The fluorescence was found in keratocyctes of all experimental groups after 4 hours,and less in the group of oligonucleotides (20μg/ml). Conclusion:Free oligonucleotides could enter the keratocytes and enhanced by liposomes