精氨酸修饰壳聚糖基因纳米粒子对血液补体活性和溶血作用的影响

Effect of arginine-modified chitosan gene nanoparticles on complement activation and hemolyzation

目的 考察精氨酸修饰壳聚糖基因纳米粒子（ACGN）对血液补体活性和溶血的影响.方法 将精氨酸接枝到壳聚糖上制备精氨酸修饰的壳聚糖,用共沉降的方法制备精氨酸修饰壳聚糖基因纳米粒子,用光子相关光谱检测精氨酸修饰壳聚糖基因纳米粒子的粒径,用凝胶电泳阻滞实验对精氨酸修饰壳聚糖与DNA的相互作用进行研究,并考察壳聚糖基因纳米粒子（CGN）和ACGN在体内外对血液补体活性和溶血作用的影响.结果 精氨酸修饰壳聚糖在电荷比为2∶1时能有效地完全包覆DNA,并形成粒径大约为120～180nm的纳米粒子,ACGN和CGN在体内外对红细胞没有明显的破坏作用,在体外对补体系统均没有明显的影响作用,但在体内引起补体的轻微升高.结论 ACGN在体内外没有引起明显的补体激活和溶血作用,有望成为一种新型的、安全的非病毒基因载体.

Objective To investigate the effect of arginine-modified chitosan gene nanoparticles （ACGN） on complement activation and hemolyzation. Methods Arginine-modified chitosan （ACS） was synthesized by incorporating arginine onto chitosan backbone. ACGN were prepared by coacervation process. Interaction between ACS and DNA was investigated by agarose gel electrophoresis retardation assay and photon correlation spectroscopy（PCS）. ELISA kits were used to investigate the effect of ACGN on complement activation and hemolyzation in vivo and in vivo. Results ACS can entrap whole plasmid DNA at charge ratio of 2：1 and formed nanoparticles with mean diameters about 120-180 nm. ACGN didn＇t play an obvious detrimental effect on red blood cells in vivo and intro. ACGN had no obvious influence on complement activation in vitro, but caused a slight increase in vivo. Conclusion ACGN didn＇t cause obvious change in complement activation and hemolyzation, which suggested that ACS would be a novel and safe non-viral vector.