Targeting delivery of oligonucleotide and plasmid DNA to hepatocyte via galactosylated chitosan vector
Targeting delivery of oligonucleotide and plasmid DNA to hepatocyte via galactosylated chitosan vector
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DOI：
10.1016/j.ejpb.2005.02.011
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发表时间：
2005-08-01
影响因子：
4.9
通讯作者：
Zhang, JF
中科院分区：
文献类型：
--
作者：
Gao, SY;Chen, JN;Zhang, JF
寡核苷酸靶向细胞并维持其生物学功能是核酸治疗的重要内容。本研究旨在证明半乳糖化低分子量壳聚糖（gal-LMWC）是一种安全有效的反义寡核苷酸（阿索）和质粒DNA肝靶向载体。MTT细胞毒性实验表明，Gal-LMWC对HepG 2细胞的毒性低于高分子量壳聚糖（HMWC）和低分子量壳聚糖（LMWC）。利用复合凝聚过程，gal-LMWC可以通过静电相互作用与质粒DNA或阿索形成稳定的纳米复合物。gal-LMWC/阿索复合物和gal-LMWC/质粒DNA复合物的形态、粒径和zeta电位非常相似。在HepG 2细胞中，使用gal-LMWC载体的转染效率显著高于裸DNA或裸阿索。gal-LMWC/阿索复合物和gal-LMWC/质粒DNA复合物的转染效率强烈地依赖于正的壳聚糖氨基和负的DNA磷酸基团的摩尔比（N/P比）。抑制实验证实，增强的转染效率是由于ASGR介导的gal-LMWC/阿索复合物或gal-LMWC/DNA复合物的内吞作用。这些结果表明，gal-LMWC可用于基因治疗，以提高转染效率在体外和体内。(c)2005年由Elsevier B. V.出版
Delivery of oligonucleotide, to specific cells and maintenance of its biological function are important for nucleic acid therapy. The objective of this paper is to demonstrate that galactosylated low molecular weight chitosan (gal-LMWC) is a safe and effective vector of antisense oligonucleotide (ASO) and plasmid DNA for the hepatocyte targeting delivery. Gal-LMWC has been successfully prepared and MTT cytotoxic assay shows that cytotoxicity of gal-LMWC is lower than that of high molecular weight chitosan (HMWC) and low molecular weight chitosan (LMWC) in HepG2 cells. Using a complex coacervation process, gal-LMWC can form stable nano-complexes with plasmid DNA or with ASO by the electrostatic interaction. The morphometrics, particle size, and the zeta potential of gal-LMWC/ASO complexes and gal-LMWC/plasmid DNA complexes are very similar. The transfection efficiency by using gal-LMWC vector is significantly higher than that of naked DNA or naked ASO in HepG2 cells. Transfection efficiency of gal-LMWC/ASO complexes and gal-LMWC/plasmid DNA complexes depends on the molar ratio of the positive chitosan amino group and the negative DNA phosphate group (N/P ratio) strongly. Inhibition experiments confirm that the enhanced transfection efficiency is due to the ASGR mediated endocytosis of the gal-LMWC/ASO complexes or gal-LMWC/DNA complexes. These results suggest that gal-LMWC can be used in gene therapy to improve the transfection efficiency in vitro and in vivo. (c) 2005 Published by Elsevier B.V.