OBJECTIVE: To prepare Aerosols containing DOTAP/DNA complexes, and to study their cell transfection efficiencies in vitro. METHODS: DOTAP/DNA complexes were prepared by modified Bligh and Dyer extraction method, which could be scattered in ether evenly with different N/P ratios. Drug-loading rate and particle size distributions were examined by inorganic phosphate assay and laser granularity distributing apparatus. Aerosols containing DOTAP/DNA complexes were further filled. The cell transfection efficiencies and cytotoxicity of aerosols were determined using human lung adenocarcinoma cell line H1299 as a model. RESULTS: The drug-loading rate of DOTAP/DNA complexes with optimal N/P ratio (2∶1) reached (72 ± 1.580)% , and the particle size was about (188.700 ± 42.770) nm. The cell survival rate after the aerosol application was (91.643 ± 6.970)% , and fluorescence intensity was 1.080×105 RLU·mg-1. CONCLUSIONS: Developed novel Aerosols containing DOTAP/DNA complexes have low toxicity and high cell transfection efficiencies, which may have the potential to be an effective gene delivery formulation to the respiratory tract.%目的:制备阳离子脂质DOTAP和DNA的复合物(DOTAP/DNA)气雾剂并研究其细胞转染效率.方法:采用改良的Bligh and Dyer萃取法,制备能够在乙醚中均匀分散的不同氮-磷比的DOTAP/DNA复合物,采用定磷法测定载药率、激光粒度分布仪测定粒径分布等参数；进一步灌装DOTAP/DNA复合物气雾剂,以人肺腺癌细胞H1299为模型,检测气雾剂的细胞毒性(细胞相对存活率)和DNA转染效率(荧光强度).结果:最佳氮-磷比(2∶1)的DOTAP/DNA复合物的载药率可达到(72±1.580)％,粒径为(188.700±42.770)nm；气雾剂的细胞相对存活率可达(91.643±6.970)％,荧光强度可达1.080× 105 RLU· mg-1.结论:研制的新型DOTAP/DNA复合物气雾剂,其毒性较低,细胞转染效率较高,具有被进一步开发研究成为呼吸道基因输送制剂的潜力.
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