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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pGL3-1us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载体。
Objective: To investigate the effect of Poloxamer 407 on the toxicity and transfection properties of polyethyleneimine (PEI). Methods: P407 was attached to the amino group of PEI by the succinimide-carbolic acid method to obtain a new polymer. The structure of the new polymer was confirmed by ~ 1H-NMR. The polymer was formed into a complex with DNA, and the complex zeta potential and MTT assay. The plasmid pGL3-1us was used as a reporter gene to determine the transfection efficiency of the luciferase activity assay complex on Hela cells. Results: ~ 1H-NMR results showed that the synthesized polymer had high purity. The Zeta potential of the complex increased with the increase of nitrogen / phosphorus ratio (N / P). The cytotoxicity of the complex increased with the increase of N / P value. The new polymer was significantly less cytotoxic than the unmodified PEI. The new polymer maintains high transfection efficiency at high N / P values. Conclusion: Poloxamer 407 modified PEI can be used as a potent non-viral gene carrier.