Jo Jun-ichiro, Ikai Tomonori, Okazaki Arimichi, Yamamoto Masaya, Hirano Yoshiaki, Tabata Yasuhiko
Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Kawara-cho Shogoin, Sakyo-ku Kyoto, Japan.
J Control Release. 2007 Apr 23;118(3):389-98. doi: 10.1016/j.jconrel.2007.01.005. Epub 2007 Jan 20.
The objective of this study is to prepare a novel gene carrier from pullulan, a polysaccharide with an inherent affinity for the liver, and evaluate the feasibility in gene transfection. Various amounts of spermine were chemically introduced into pullulan with molecular weights of 22,800, 47,300, and 112,000 to prepare cationized pullulan derivatives with different percentages of spermine introduced. Each cationized pullulan derivative was complexed with a plasmid DNA at various ratios and applied to HepG2 cells for in vitro gene transfection. The level of gene expression depended on the percent spermine introduced of cationized pullulan derivatives and the molecular weight of pullulan. However, when compared at the complexation molar ratio of pullulan derivative to the plasmid DNA, the expression level became maximum around the ratio of 10(2), irrespective of the pullulan molecular weight. Pre-treatment of cells with asialofetuin of asialoglycoprotein receptor ligand decreased the level of gene expression by the complexes. The cationized pullulan derivative with an appropriate physicochemical character is a promising non-viral carrier which promotes the receptor-mediated internalization of plasmid DNA and consequently enhances the expression level.
本研究的目的是利用对肝脏具有内在亲和力的多糖支链淀粉制备一种新型基因载体，并评估其在基因转染中的可行性。将不同量的精胺化学引入分子量分别为22,800、47,300和112,000的支链淀粉中，以制备精胺引入百分比不同的阳离子化支链淀粉衍生物。每种阳离子化支链淀粉衍生物与质粒DNA以不同比例复合，并应用于HepG2细胞进行体外基因转染。基因表达水平取决于阳离子化支链淀粉衍生物中精胺的引入百分比和支链淀粉的分子量。然而，当以支链淀粉衍生物与质粒DNA的复合摩尔比进行比较时，无论支链淀粉分子量如何，在10(2)左右的比例时表达水平达到最高。用去唾液酸糖蛋白受体配体去唾液酸胎球蛋白预处理细胞可降低复合物的基因表达水平。具有适当理化性质的阳离子化支链淀粉衍生物是一种有前途的非病毒载体，可促进质粒DNA的受体介导内化，从而提高表达水平。
