弹性阳离子脂囊泡载入的人酪氨酸酶基因表达和黑色素生成的增强。
Enhancement of gene expression and melanin production of human tyrosinase gene loaded in elastic cationic niosomes.
机构信息
Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand.
出版信息
J Pharm Pharmacol. 2012 Oct;64(10):1376-85. doi: 10.1111/j.2042-7158.2012.01509.x. Epub 2012 Mar 22.
OBJECTIVES
Disturbance in the synthesis of tyrosinase might be one of the major causes of vitiligo. The enhancement of tyrosinase gene expression and melanin production by loading the plasmid in elastic cationic niosomes was investigated in tyrosinase gene knocked out human melanoma (M5) cells and in tyrosine-producing mouse melanoma (B(16) F(10) ) cells.
METHODS
Niosomes composed of Tween 61/cholesterol/dimethyl dioctadecyl ammonium bromide at 1:1:0.5 molar ratio were prepared by the freeze-dried empty liposomes method. The thin lipid film was redissolved in distilled water or 25% ethanol to obtain the non-elastic or elastic cationic niosomes, respectively.
KEY FINDINGS
The maximum loading of the plasmid in non-elastic and elastic niosomes was 130 and 100 µg per 16 mg of the niosomal contents, respectively. The plasmid-loaded elastic cationic niosomes exhibited high specific tyrosinase activity of 1.66 and 1.50 fold in M5 cells and 6.81 and 4.37 fold in B(16) F(10) cells compared with the free plasmid and the plasmid-loaded non-elastic cationic niosomes, respectively.
CONCLUSIONS
This study has demonstrated not only the enhancement of the expression of human tyrosinase gene by loading in elastic cationic niosomes, but also the potential application of this gene delivery system for the further development of vitiligo gene therapy.
目的
酪氨酸酶合成障碍可能是白癜风的主要原因之一。本研究旨在探讨将质粒载入具有弹性正离子的阳离子脂囊泡后对人黑色素瘤(M5)细胞和产酪氨酸的鼠黑色素瘤(B(16)F(10))细胞中天冬氨酸酶基因表达和黑色素生成的影响。
方法
采用冻干空白脂囊泡法制备由吐温 61/胆固醇/二甲基双十八烷基溴化铵以 1:1:0.5 摩尔比组成的脂囊泡。将薄脂膜重新溶解于蒸馏水中或 25%乙醇中,分别得到非弹性或弹性阳离子脂囊泡。
主要发现
非弹性和弹性脂囊泡对质粒的最大载入量分别为 130 和 100μg/16mg 脂囊泡内容物。与游离质粒和载入非弹性阳离子脂囊泡的质粒相比,载入弹性阳离子脂囊泡的质粒在 M5 细胞中表现出的天冬氨酸酶比活性分别增加了 1.66 倍和 1.50 倍,在 B(16)F(10)细胞中分别增加了 6.81 倍和 4.37 倍。
结论
本研究不仅证实了将质粒载入弹性阳离子脂囊泡后可增强人酪氨酸酶基因的表达,还为进一步开发白癜风基因治疗提供了这种基因传递系统的潜在应用价值。