多聚体超声造影剂微泡携带DNA能力研究

Ability of ultrasound contrast agent microbubles to carry DNA

目的 研究自行研制的新型多聚体超声造影剂(UCA)微泡携带DNA片段的能力。方法 以乳液共聚法制备多聚体UCA;以人血细胞提取人类基因组DNA,并用超声将其破碎为DNA片段;将多聚体UCA与PI染色的DNA片段共孵育30、60、90、120分钟后,在荧光显微镜下观察微泡与DNA片段结合情况;将多聚体UCA与PI染色的DNA片段共孵育30分钟,经不同转速离心后观察微泡与DNA片段结合情况。结果 多聚体UCA与PI染色的DNA片段共孵育30分钟后,荧光显微镜下均可见带有红色荧光物质的微泡分布,没有发现由于孵育时间延长,微泡表面荧光增强或减弱的现象,不同离心转速对微泡表面荧光强度没有影响。结论 自行研制的新型多聚体UCA微泡与DNA片段在30分钟以内即可结合,并经过高速离心仍然保持与DNA片段结合状态。

Objective To study the ability of a kind of new self-designed polymer ultrasound contrast agent (UCA) microbubble to carry DNA. Methods The self-designed polymeric UCA were prepared by means of emulsion copolymerization. Human genome DNA, which was extracted from human blood cells, was broken into small fractions by ultrasound. The polymeric UCA were incubated with PI-labeled DNA fractions 30, 60, 90 and 120 minutes, then observed separately under fluorescence microscopy. After polymer UCA were incubated with PI-labeled DNA fractions 30 minutes, it was take into a centrifugal machine at different velocities. Fluorescence microscopy was used again to compare the two conditions. Results After the polymeric UCA were incubated with PI-labeled DNA fractions 30 minutes, microbubles were found with red fluorescent light under fluorescence microscopy. With the incubating time extending, there was not stronger red light fluorescent than ever before. The red light fluorescent did not change with different velocities of centrifugal machine. Conclusion The self-designed polymeric UCA can be combined with DNA fractions in 30 minutes. High-velocity centrifugal effect does not change the combination.