透明质酸键合分枝状低分子量聚乙烯亚胺作为siRNA载体的研究  

Hyaluronic Acid-Graft-Branch Low Molecular Weight Polyethylenimine for Gene Delivery

作　　者：唐川[1] 刘俊成 蒋革[1] TANG Chuan;LIU Juncheng;JIANG Ge(College of Life Science and Technology,Dalian University,Dalian 116622,China)

机构地区：[1]大连大学生命科学与技术学院,辽宁大连116622

出　　处：《重庆理工大学学报（自然科学）》2020年第4期209-214,共6页Journal of Chongqing University of Technology：Natural Science

基　　金：国家自然科学基金青年科学基金项目(21606030);辽宁省自然科学基金指导计划(201602020);2019年大连市高层次人才创新项目支持计划。

摘　　要：通过构建透明质酸键合低分子量聚乙烯亚胺的低毒性siRNA载体,探究其转染si RNA的效果。利用核磁共振氢谱确定透明质酸与低分子量聚乙烯亚胺嫁接形成复合物的结构,并通过MTT方法检测其对细胞的毒性。采用凝胶电泳法确定siRNA与复合载体的结合比,利用粒度仪对其粒径大小、带电量等进行表征,并通过透射电子显微镜观察其形态。通过复合载体在B16F1细胞内,观察其基因沉默效率。结果表明:当siRNA与复合载体的质量比为1∶20时,其表面带电量约为+0. 3 m V,粒子直径约为113. 7 nm,形态均匀呈球形,分散均匀。其对细胞毒性远远小于高分子量聚乙烯亚胺,但转染效果较好。因此,透明质酸键合低分子量聚乙烯亚胺是一种低毒、高效的新型非病毒载体。To explore the transfection effect of hyaluronic Acid-Bonded low-molecular-weight polyethyleneimine with low-toxicity si RNA vector,the structure of hyaluronic acid-low molecular weight polyethyleneimine grafted complex was determined by 1 H NMR,and its cytotoxicity was measured by MTT assay. The binding ratio of siRNA to the composite carrier was determined by gel electrophoresis. The particle size and charged volume were characterized by particle size analyzer,and the morphology was observed by transmission electron microscope. The efficiency of gene silencing in B16 F1 cells was observed by using complex vectors. The results showed that,when the mass ratio of siRNA to composite carrier is 1∶ 20,the surface charged amount is about + 0. 3 mV,the particle diameter is about 113. 7 nm,and the morphology is spherical and uniform. Its cytotoxicity is far less than that of high molecular weight polyethyleneimine,but its transfection effect is better.Therefore,hyaluronic acid bonded low molecular weight polyethyleneimine is a new non-viral vector with low toxicity and high efficiency.

关 键 词：透明质酸 低分子量 聚乙烯亚胺 siRNA载体 转染 

分 类 号：R318.08[医药卫生—生物医学工程]