负载MMP-2 siRNA阳离子脂质体的制备及体外转染肝星状细胞的研究.docVIP

 负载 MMP-2 siRNA 阳离子脂质体的制备及 体外转染肝星状细胞的研究# 刘凤，陈平圣** 5 10 15 20 25 30 35 （东南大学医学院病理学与病理生理学系，南京 210009） 摘要：目的：研制负载基质金属蛋白酶-2（matrix metalloproteinase-2, MMP-2）siRNA 的阳 离子脂质体，对其体外理化性质进行检测并观察对肝星状细胞（hepatic stellate cells, HSC） 的毒性作用、转染效率及基因沉默效率。方法：采用薄膜分散—高压均质法制备阳离子脂 质体，用透射电镜、激光粒度分析仪对其进行表征，体外转染 HSC-T6，评价其细胞毒性、 转染效率及基因沉默效率。结果：本法制备的阳离子脂质体平均粒径为 133.9nm，分散性良 好，呈球形或卵球形，其平均 zeta 电位为 41.67mV。体外作用于 HSC-T6，细胞毒性低，转 染效率为 72.4%，三条 MMP-2 siRNA 基因沉默效率分别为 50.6%、40.0%、53.4%。结论： 采用上述方法制备的负载 MMP-2 siRNA 的阳离子脂质体，粒径较小，具有较好的稳定性， 在保证转染效率的同时，对 HSC-T6 细胞毒性较小，基因沉默效率高，从而为 MMP-2 siRNA 干扰实验研究提供了一个有效的工具。 关键词：病理学；阳离子脂质体；siRNA；肝星状细胞；基质金属蛋白酶-2；转染效率 中图分类号：R943；R365 Preparation of cationic liposomes delivering MMP-2 siRNA and its transfection study of hepatic stellate cells in vitro LIU Feng1, CHEN Pingsheng2 (1. Medical School, Southeast University，Nanjing 210009; 2. Medical School,Southeast University, Nanjing 210009) Abstract: Objective: To explore the preparation method and characteristics of cationic liposomes delivering MMP-2 siRNA and evaluate its toxicity, transfection efficiency and gene silencing efficiency on hepatic stellate cells. Methods: The cationic liposomes were prepared by film dispersion and high pressure homogenizer method. TEM, ZETA-SIZER were used to characterize cationic liposomes. Transfect HSC in vitro and evaluate its toxicity, transfection activity and gene silencing efficiency. Results: The average diameter of cationic liposomes was about 133.9 nm and the average zeta potential was 41.67mV. Transfect HSC in vitro showed that it has low cytotoxicity, high transfection activity, and the three siRNA of gene silencing efficiency is50.6%、 40.0%、53.4%. Conclusion: Cationic liposomes which have smaller particle size, good stability, low toxicity and high gene silencing efficiency can be successfully prepared by these methods. This study provides an effective tool to siRNA interference experiments. Key words: pathology; cationic liposomes; siRNA; hepatic stellate cells; matri