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目的 对不同方法制备酸敏脂质体进行包封率测定及形态学观察 ,探讨酸敏脂质体包裹反义寡核苷酸的生物学效应。方法 测定四种不同方法制备包裹1 2 5I IL 8脂质体的包封率 ;在超高倍显微分析仪下观察形态结构。用包裹细胞磷脂酶A2 (cPLA2 )反义寡核苷酸的酸敏脂质体转染U937细胞 ,提取细胞RNA ,采用反转录 聚合酶链反应 (RT PCR) ;免疫印迹 (Westernblot)法检测cPLA2 蛋白的表达。结果 反相蒸发法与钙融合法联合制备的酸敏脂质体具有良好地包封率和形态结构 ,经酸敏脂质体包裹cPLA2 反义寡脱氧核苷酸可明显抑制cPLA2基因及蛋白的表达。结论 制备所得的酸敏脂质体可良好的发挥其生物学效应
OBJECTIVE To evaluate the entrapment efficiency and morphology of acid-sensitive liposomes prepared by different methods, and to investigate the biological effects of acid-sensitive liposome-coated antisense oligonucleotides. Methods The entrapment efficiency of 1 215 IL 8 liposomes prepared by four different methods was determined. Morphology was observed under ultra-high magnification microscopy. U937 cells were transfected with acid liposomes encapsulating the cellular phospholipase A2 (cPLA2) antisense oligonucleotides, and RNA was extracted by reverse transcription polymerase chain reaction (RT PCR). Western blotting cPLA2 protein expression. Results Acid-sensitive liposomes prepared by reverse-phase evaporation combined with calcium-fusing method had good entrapment efficiency and morphological structure. The inclusion of cPLA2 antisense oligodeoxynucleotides could significantly inhibit the cPLA2 gene and protein expression. Conclusion The prepared acid-sensitive liposomes can exert their biological effects well