| 摘要: |
| 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。 |
| 关键词: PCM 脂质体 质粒 心脏肌细胞 转染效率 |
| DOI：10.16781/j.0258-879x.2016.11.1348 |
| 投稿时间：2016-03-08修订日期：2016-08-31 |
| 基金项目:国家自然科学基金（81400212）. |
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| Preparation of PCM modified liposome and evaluation of its myocardium targeting efficiency |
| WANG Xin,HUANG Hua,WANG Mou,CHEN Hua-li* |
(Chongqing Medical Engineering Research Center, Chongqing Medical University, Chongqing 400016, China
*Corresponding author.) |
| Abstract: |
| Objective To prepare PCM modified liposome (PCM-LIP) containing enhanced green fluorescent protein expression plasmid (pEGFP) and to evaluate its myocardial targeting ability. Methods Liposome was prepared by film-ultrasonic, with PCM used as ligand and DOTAP as cationic lipid material. PCM-LIP containing pEGFP was prepared by incubating liposome with pEGFP at room temperature. The connecting method of PCM was optimized and the connection rate of PCM was determined. The characteristics of liposomes including encapsulation ability, morphology, particle size, zeta potential and stability in phosphate buffer solution (PBS) were observed. The transfection efficiency of liposomes into H9c2 cells was evaluated by inverted fluorescence microscopy and flow cytometry, so as to characterize their myocardial targeting ability and to determine the optimum dosage of PCM. Results PCM-LIP was prepared by insertion method and the amount of PCM accounted for 3% of the lipid. After incubation with pEGFP, PCM-LIP was spherical in shape, with the particle size being (261.9±2.2) nm, zeta potential being (-5.0±0.6) mV, and PCM-LIP was stable in PBS at -4℃ for 30 d. The transfection efficiency of PCM-LIP was significantly higher than that of unmodified liposome (P<0.05). Conclusion PCM can improve the transfection efficiency of liposome into cardiomyocytes and PCM-LIP shows a satisfactory myocardial targeting ability. |
| Key words: PCM liposomes plasmids cardiac myocytes transfection efficiency |
