阳离子胆固醇的合成及其基因传递性能研究

发布时间：2018-10-16 22:13

【摘要】：采用琥珀酸将N,N-二甲基乙二氨偶联到胆固醇上,合成新型的阳离子胆固醇衍生物——N-(N′,N′-二甲基)乙基丁二酸单胆固醇酯单酰胺(DC-CHEMS),并考察其作为基因载体的可行性.以DC-CHEMS和磷脂为膜材采用薄膜分散法制备脂质体,透射电镜显示其形态规整,粒径均匀;激光粒度测定仪测得其平均粒径为136nm,Zeta电位为25mV.琼脂糖凝胶电泳显示当+/-电荷比大于4∶1时,脂质体能将DNA完全结合,且能保护DNA免受DNase I酶的降解.溶血试验和MTT细胞毒性试验结果显示,所得脂质体具有较低的毒性.细胞摄取和基因转染实验表明:所得脂质体能显著促进HepG2细胞对F-ODN的摄取;当+/-电荷比为1∶1,1∶2时,基因转染效果强于市售的转染试剂Lipfectamine汶2000.因此,DC-CHEMS作为基因载体,具备了高效低毒的优势,值得进一步的深入研究.
[Abstract]:A new kind of cationic cholesterol derivative N- (N-) -dimethyl) ethyl succinate monoamide (DC-CHEMS) was synthesized by using succinic acid (Succinic acid) to couple N-dimethyl-ethylenediamine to cholesterol. The feasibility of N- (N-) -dimethyl-ethylenediamine (DC-CHEMS) as a gene carrier was investigated. Liposomes were prepared by membrane dispersion method using DC-CHEMS and phospholipid as membrane materials. The morphology of liposomes was uniform by transmission electron microscope, and the average particle size of liposomes was 136nmPe (25mV) measured by laser particle size analyzer. Agarose gel electrophoresis showed that when the /-charge ratio was greater than 4:1, the liposome could bind DNA completely and protect DNA from DNase I degradation. The results of hemolysis test and MTT cytotoxicity test showed that the liposomes had lower toxicity. The results of cell uptake and gene transfection showed that the liposomes could significantly promote the uptake of F-ODN in HepG2 cells, and when the ratio of / to charge was 1: 1: 1: 2, the effect of gene transfection was better than that of the marketable transfection reagent Lipfectamine Wen2000. Therefore, as a gene vector, DC-CHEMS has the advantages of high efficiency and low toxicity, which is worthy of further study.
【作者单位】：浙江工业大学药学院;
【基金】：国家自然科学基金资助项目(81102396)
【分类号】：R944

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