精氨酸修饰壳聚糖基因纳米粒子对血小板GMP-140表达的影响

目的 考察精氨酸修饰壳聚糖基因纳米粒子(ACGN)对血小板GMP-140表达的影响.方法 制备精氨酸修饰的壳聚精(ACS)并朋红外光谱对其进行表征;用共沉降的方法制备精氨酸修饰壳聚糖基因纳米粒子,用凝胶电泳阻滞实验对精氨酸修饰壳聚精与DNA的相互作用进行研究;用酶联免疫双抗夹心法测定血小板α颗粒膜蛋白-140(GMP-140)表达来考察精氨酸修饰壳聚糖基因纳米粒子对血小板激活的影响.结果 红外光谱结果显示精氨酸成功地接枝到壳聚糖上,在正负电荷比≥2:1时,ACS能完全阻滞DNA的迁移,表明所有的DNA均已被ACS完全包覆.壳聚糖基因纳米粒子(CGN)和ACGN在体外均不引起血小板的激活,但在体内则都引起轻微血小板激活.结论 精氨酸修饰壳聚糖纳米粒子对血小板GMP-140的表达没有引起明显的变化,有望成为一种新型的、安全的非病毒基因载体.

Effect of arginine-modified chitosan gene nanoparticles on the change of platelet GMP-140

Objective To investigate the effect of arginine-modified chitosan gene nanoparticles(ACGN)on the change of platelet GMP-140. Methods Arginine-modified chitosan(ACS) was synthesized and characterized by FTIR. ACGN were prepared by coacervation process. Interaction between ACS and DNA was investigated by agarose gel electrophoresis retardation assay. GMP-140 ELISA kits were used to investigate the effect of arginine-modified chitosan gene nanoparticles on the change of platelet GMP-140 in vivo and in vivo. Results FTIRresult demonstrated that arginine was successfully incorporated into the backbone of chitosan. Plasmid DNA was entirely entrapped by ACS at the charge ratio of 2:1 or above. Platelet activation wasn't caused by CGN or ACGN in vitro, but slight platelet activations were caused by both CGN and ACGN in vivo, as compared with the control group. Conclusion ACS didn't cause obvious change in platelet GMP-140, which suggested that ACS would be a novel and safe non-viral vector.