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目的合成非病毒基因载体材料聚谷氨酸-聚丙烯亚胺,并考查其与DNA的结合能力及稳定性。方法以G2.0聚丙烯亚胺(PPI)为引发剂,通过开环聚合的方法,合成了聚丙烯亚胺-聚谷氨酸苄脂多臂接枝聚合物,再以G1.0聚丙烯亚胺或G2.0聚丙烯亚胺作为氨解试剂进行氨解合成了新型树枝状的聚谷氨酸-聚丙烯亚胺聚合物(MP-g-PPI)。采用红外光谱(IR)、1HNMR对聚合产物进行结构表征,通过琼脂糖凝胶电泳实验考察聚合物与DNA的结合能力,通过离心法考察了聚合物与DNA形成复合物的沉淀稳定性,通过MTT法考察聚合物在Hela细胞中的毒性,并通过测定发荧光细胞百分数获得复合物在Hek-293细胞中的转染率。结果成功合成了聚谷氨酸-聚丙烯亚胺聚合物,此聚合物可有效的与DNA结合成纳米复合物,可通过一定的空间斥力阻止复合物的聚集,且毒性较PEI 25K低,可有效介导基因的转染。结论聚谷氨酸-聚丙烯亚胺聚合物可与DNA形成稳定的复合物,此聚合物有望成为新型的非病毒基因载体材料。
OBJECTIVE: To synthesize polyglutamic acid-polypropyleneimine (non-viral gene carrier material) and investigate its binding ability and stability with DNA. Methods Polypropyleneimine-polyglutamic acid benzyl ester multi-arm graft polymer was prepared by ring-opening polymerization using G2.0 polypropyleneimine (PPI) as initiator, The new dendritic polyglutamic acid-polypropyleneimine polymer (MP-g-PPI) was synthesized by aminolysis of imine or G2.0polyacrylimide as aminolysis reagent. The polymer was characterized by IR and 1HNMR. The binding ability between the polymer and DNA was investigated by agarose gel electrophoresis. The precipitation stability of polymer-DNA complex was investigated by MTT method. Method to examine the toxicity of the polymer in Hela cells, and the transfection efficiency of the complex in Hek-293 cells was obtained by measuring the percentage of fluorescing cells. Results Polyglutamic acid-polypropyleneimine polymer was synthesized successfully. This polymer can be effectively combined with DNA into nanocomposite. It can block the aggregation of the compound through a certain space repulsion, and its toxicity is lower than that of PEI 25K. Effectively mediate gene transfection. Conclusion Polyglutamic acid-polypropyleneimine polymer can form a stable complex with DNA. This polymer is expected to become a new non-viral gene carrier material.