生物还原响应的葡聚糖修饰的线性聚乙烯亚胺基因释放载体的研究

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王春晓1， 林超2， 赵鹏2， 赵健1*

 (1.青岛科技大学 橡塑材料与工程教育部重点实验室/橡塑材料与工程山东省重点实验室，山东 青岛 266042；2.同济大学 生物医学工程与纳米科学研究院，上海 200092)

摘要： 通过二硫吡啶端基化的线性聚乙烯亚胺（LPEISSPy,Mn=5 000）与巯基化葡聚糖（DexSH,Mw=5 000）的偶联反应，制备出生物还原响应的葡聚糖基线性聚乙烯亚胺（DexSSLPEI），并将其用于基因释放载体的研究。核磁共振氢谱分析表明DexSSLPEI的氢谱与其化学结构一致。动态光散射及琼脂糖凝胶电泳分析证明DexSSLPEI能够复合基因形成纳米尺寸的、表面带正电荷的复合物。毒性实验表明该聚合物对细胞的毒性较小。在n(N)/n(P)为最佳值(10/1)时，DexSSLPEI与绿色荧光蛋白报告基因的复合物可以有效转染MCF7和HepG2细胞，其效率优于22 kDa的线性聚乙烯亚胺（LPEI,Mw=22 kDa）阳性对照。荷瘤小鼠体内转染实验表明DexSSLPEI可以通过静脉给药输送荧光素酶基因，导致体内各器官及肿瘤的转基因表达。在肿瘤内该转基因表达较其它脏器更高，并高于22 kDa的LPEI在肿瘤内的基因表达。

 关键词： 线性聚乙烯亚胺； 葡聚糖； 基因载体； 绿色荧光蛋白

 中图分类号： R 318     文献标志码： A

 Bioreductionresponsive Dextranated Linear Polyethylenimine as Gene Delivery Vector

WANG Chunxiao1， LIN Chao2， ZHAO Peng2， ZHAO Jian1

 (1.Key Laboratory of RubberPlastics, Ministry of Education/Shandong Provincial Key Laboratory of RubberPlastics, Qingdao University of Science and Technology, Qingdao 266042, China；2.The Institute for Biomedical Engineering and Nano Science, Tongji University, Shanghai 200092, China)

Abstract: By conjugating linear polyethylenimine dithiopyridine (Mn=5 000) with thiolated dextran （DexSH,Mw=5 000）, a group of disulfidelinked dextranLPEI conjugates (DexSSLPEI) are prepared and evaluated as nonviral vectors for gene delivery. 1H NMR spectra of these DexSSLPEI conjugates are in line with their chemical structure. Dynamic light scattering and argarose gel analysis manifest that the conjugates can bind gene to form nanoscale polyplexes. Cytotoxicity test show that DexSSLPEI conjugates have low cytotoxicity. At an optimal n(N)/n(P) ratio of 10/1, the polyplexes of DexSSLPEI conjugates with the plasmid encoding green fluorescence protein (pCMVGFP) can efficiently transfect MCF7 and HepG2 cells, with the efficiency which is higher than that yielded by 22 kDa LPEI as a positive control. Moreover, the conjugates are applicable for intravenous delivery of the plasmid encoding luciferase (pCMVLuc) into HepG2 tumorbearing nude mice, causing transgene expression largely in tumor rather than other organs. Moreover, the conjugates lead to higher transgene expression in the tumor than 22 kDa LPEI as a positive control.

Key words: linear polyethylenimine; dextran; gene delivery vector;green fluorescence protein

收稿日期： 20141009

基金项目： 国家自然科学基金项目（51073082；51373088).

作者简介： 王春晓（1986—），女，硕士研究生. *通信联系人.

文章编号： 16726987（2015）06060507； DOI： 10.16351/j.16726987.2015.06.003