摘要
目的研究阳离子脂质体膜的组分对细胞摄取和基因转染的影响。方法以十八酰胺(SA)3、β[N-(N′,N′-二甲基氨基乙基)氨甲酰基-胆固醇(DC-Chol)、卵磷脂(SPC)、二油酰磷脂酰乙醇胺(DOPE)等为膜材,制备阳离子脂质体。用流式细胞仪测定不同阳离子脂质体对细胞摄取荧光标记寡核苷酸(FAM-ODN)的效率,结合荧光显微镜观察报告基因(pEGFP-N1)编码的绿荧光蛋白(GFP)表达的情况。结果不同阳离子脂质对细胞摄取产生截然不同的效果,DC-Chol/SPC脂质体对细胞摄取ODN的促进作用显著高于SA/SPC脂质体;而中性脂质对基因表达有重要影响,DC-Chol/DOPE脂质体的转染效率优于DC-Chol/SPC脂质体。结论通过药剂学的处方筛选,可以提高阳离子脂质体作为非病毒载体的性能。
OBJECTIVE To study the effect of the lipid compositions on cellular uplake and gene transfection mediated by cationic liposomes. METHODS Various lipids such as stearylamine （SA）, 3β[ N- （ N＇, N＇-dimethylaminoethan）-carbamoyl] cholesterol （DC-Chol）, soy bean phosphatidylcholine （SPC） and dioleoyl phosphatidylethanolaminc （DOPE） were selected to prepare cationic liposomes. The cellular uptake of fluorescence labeled oligonucleotides （FAM-ODN） was determined by flow cytometric analysis. The expression of plasmid enceded green fluorscence proteins （GFP） was observed under fluorescence microscopy and analyzed by flow cytometer. RESULTS The cellular uptake efficiency was significantly influenced by the type of cationic lipid. DC-Chol/SPC liposomes influenced more efficiently in enhancing cellular uptake than SA/SPC liposomes. However, DC-Chol/DOPE liposomes showed high transfection efficiency than DC-Chol/SPC liposomes, indicating that the neutral lipid played a key role in the process of gene transfection. CONCLUSION These results demonstrate that utilization of pharmaceutical technology may improve the gene delivery based on cationic liposomes.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2006年第3期232-235,共4页
Chinese Pharmaceutical Journal
基金
国家自然科学基金资助项目(30371692)
浙江省自然科学基金资助项目(Y205451)
关键词
阳离子脂质体
细胞摄取
基因转染
脂质组分
cationic liposomes
cellular uptake
gene transfection
lipid compositions
