泊洛沙姆407修饰对聚乙烯亚胺的毒性和转染性质的影响

目的:考察泊洛沙姆407修饰对聚乙烯亚胺(PEI)的毒性和转染性质的影响.方法:使用琥珀酰亚胺碳酸脂法将P407连接在PEI的氨基上,得到新聚合物,通过1H-NMR确定新聚合物的结构,将该聚合物与DNA形成复合物,测定复合物的zeta电位,MTT法考察复合物的细胞毒性,使用质粒pGL3-lus作为报告基因,测定虫荧光素酶活性评价复合物对Hela细胞的转染效率.结果:1H-NMR结果表明合成的聚合物具有较高的纯度.复合物的Zeta电位随氮/磷比(N/P)值的增加而增高.复合物的细胞毒性随着N/P值的增加而增大,新聚合物其细胞毒性显著低于未修饰的PEI.新聚合物在高N/P值时仍能保持较高的转染效率.结论:泊洛沙姆407修饰的PEI可以作为一种有效的非病毒基因栽体.

Effects of Poloxamer 407 Modification on the Cytotoxicity and Transfection Activity of Polyethyleneimine

Objective ： To study the effect of poloxamer 407 （P407） modification on the cytotoxicity and transfection activity of polyethyleneimine （PEI）. Method： PEI was modified by conjugating PEI with P407 with suceinimidyl carbonate method. Structural a- nalysis of the synthesized polymer was performed by 1H-NMR. Zeta potential of the synthesized polymer/DNA complexes was measured. The cytotoxicity of the complexes was evaluated using MTT method. The pGL3-1us as the reporter gene, the transfection efficiency of the complexes was determined in vitro by measuring luciferase activity in Hela cells. Result： The result of 1H-NMR showed the purity of the synthesized polymers was high. Zeta potential and the cytotoxicity of the complexes were both increased with the increase of N/P ratios. The synthesized polymers showed lower cytotoxicity than the unmodified PEI. The synthesized polymers at high N/P ratios still showed high transfection efficiency. Conclusion： The P407-modified PEI can be an effective non-viral genetic carder.