酸敏脂质体的制备及其生物学活性

Preparation and biological activity of pH-sensitive liposome

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。

Objective To measure the envelo pm ent rate and observe the conformation of pH-sensitive liposome made by differe nt methods,to check the biological activity of pH-sensitive liposome-wrapped a ntisense oligodeoxynucleotide (ASODN).Methods Liposome-capsulated 125 I-IL-8 mad e by four kinds of methods were compared for their envelopment rates,their confo rmation was observed with high-power microscopic analyzer;using pH-sensitive l iposome-wrapped cell phospholipase A 2 (cPLA 2) to transfect the U937 cell.RN A of cell was extracted,expression of cPLA 2 protein was checked with Western b lot.Results The liposome made by calcium-induced fusion and revers e-phase evaporation has the highest envelopment rate and the best conformation .pH-sensitive liposome-wrapped cPLA2 can restrain the expression of gene and p rotein.Conclusion The pH-sensitive liposome made by the author has a good biological activity.