胎牛血清对聚乙烯亚胺基因传递系统的降毒作用研究

Study on Toxicity Attenuation Effects of Fetal Bovine Serum on Polyethylenimine Gene Delivery System

目的:研究胎牛血清(FBS)对聚乙烯亚胺(PEI)基因传递系统的降毒作用。运用Nanoparticles albumin bound技术的原理,制备以FBS修饰PEI/DNA的复合物(PEI/DNA/FBS)。采用MTT比色法计算人宫颈癌上皮Hela细胞存活率(CV),测定PEI质量浓度分别为0.5、1、2、5、10、15、20、30μg/ml时的细胞毒性,比较FBS含量分别为0、2.5%、5%、10%时PEI/FBS复合物和N(PEI中氮的物质的量)/P(DNA中磷的物质的量)比分别为10、20时的PEI/DNA/FBS复合物的细胞毒性差异,比较PEI和N/P比分别为10、20且FBS含量为0、5%、10%的PEI/DNA/FBS复合物作用4、8、12、16 h时的细胞毒性差异。结果:CV值与PEI质量浓度呈负相关,当PEI≥10μg/ml时,对细胞有明显的毒性;随FBS含量的增加CV值增加,当FBS含量为2.5%、5%、10%时PEI/DNA/FBS复合物CV值均在80%以上,且FBS含量为5%、10%时CV值均大于100%;且随作用时间的延长CV值基本维持稳定。结论:FBS可有效降低PEI基因传递系统的细胞毒性,经FBS修饰后的PEI基因传递系统不会随作用时间的延长而加大对细胞的损伤。

OBJECTIVE： To study the toxicity attenuation effects of fetal bovine serum （FBS） on the polyethyleneimine （PEI） gene delivery system. METHODS： Nanoparticles albumin bound technology was conducted to prepare the FBS-modified PEI/DNA compound PEI/DNA/FBS. MTT assay was used to calculate the Hela cell viability rate （CV） in human cervical epithelium, deter- mine the cytotoxicity of PEI with the mass concentration of 0.5, 1, 2, 5, 10, 15, 20 and 30μg/ml, compare the toxicities be- tween the PEI/FBS with the FBS content of 0, 2.5%, 5% and 10% and PEI/DNA/FBS with the N （amount of nitrogen in PEI）/P （amount of phosphorus in DNA） ratio of 10 and 20, and compare the toxicities changes between PEI and PEI/DNA/FBS with the N/P ratio of 10 and 20 for 4, 8, 12 and 16 h, respectively. RESULTS： CV value and PEI mass concentration were negatively corre- lated and it had obvious toxicity to cells when PEI ≥ 10μg/ml; the CV value was increased with the increasing of FBS content, the CV value of compound PEIfDNA/FBS was more than 80% when the FBS content was respectively 2.5%, 5% and 10% and the CV value was more than 100% when the FBS contents were respectively 5% and 10% ; and the CV value was basically stable with the prolonging of action time. CONCLUSIONS： FBS can effectively reduce the cell toxicity of PEI gene delivery system and the FBS-modified PEI gene delivery system will not increase the cell damage with the prolonging of action time.