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
Preparation of cationic liposomes delivering MMP-2 siRNA and its transfection study of hepatic stellate cells in vitro
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。
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.
刘凤、陈平圣
基础医学生物科学研究方法、生物科学研究技术
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
pathologycationic liposomessiRNAhepatic stellate cellsmatrix metalloproteinase-2transfection activity
刘凤,陈平圣.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[EB/OL].(2012-03-31)[2025-08-06].http://www.paper.edu.cn/releasepaper/content/201203-857.点此复制
评论