论文部分内容阅读
目的制备心肌特异性靶向肽PCM修饰的载增强绿色荧光蛋白表达质粒(pEGFP)的脂质体(PCM-LIP),并初步考察其心肌靶向性。方法采用薄膜分散-超声法,以PCM为配体,DOTAP为阳离子脂质材料制备脂质体,PCM-LIP与pEGFP室温孵育制备载质粒的脂质体。对PCM连接方法进行优化,测定PCM的连接率,并考察脂质体的载药能力、形态、粒径分布、电位及其在磷酸盐缓冲液(PBS)中的稳定性。通过倒置荧光显微镜和流式细胞术考察脂质体转染心肌细胞H9c2的效果,表征其心肌靶向性,筛选PCM的最佳用量。结果 PCM通过插入法连接,用量为脂质的3%,与pEGFP孵育后的PCM-LIP呈圆球形,粒径为(261.9±2.2)nm,zeta电位在为(-5.0±0.6)mV,在PBS中具有良好的稳定性,其对心肌细胞的转染效率高于未修饰的脂质体(P<0.05)。结论 PCM可提高脂质体对心肌细胞的转染效率,PCM-LIP具有良好的心肌细胞靶向性。
OBJECTIVE: To prepare PCM-LIP (PCM-LIP) modified with PCM-specific enhancer of pEGFP and to investigate its cardiac targeting. Methods Liposomes were prepared by membrane dispersion-sonication with PCM as ligand and DOTAP as cationic lipid. PCM-LIP and pEGFP were incubated at room temperature to prepare liposomes. The PCM connection method was optimized to determine the connectivity of PCM. The drug loading ability, morphology, particle size distribution, potential and its stability in phosphate buffered saline (PBS) were investigated. The effects of liposome transfection on H9c2 cells were investigated by inverted fluorescence microscopy and flow cytometry, and the optimal targeting of PCMs was obtained. Results The PCM was inserted into the medium at an amount of 3% of the lipid. After incubation with pEGFP, the PCM-LIP was spherical with a diameter of (261.9 ± 2.2) nm and a zeta potential of (-5.0 ± 0.6) mV. PBS has good stability, its transfection efficiency of myocardial cells than unmodified liposomes (P <0.05). Conclusions PCM can enhance the transfection efficiency of liposome to cardiomyocytes, and PCM-LIP has good cardiomyocyte targeting.