液态氟碳脂质微球靶向损伤心肌细胞的体外实验

In Vitro Experimental Study of Rat Cardiomyocyte Injury with Targeting of Perfluorocarbon Lipid Particles

采用生物素-亲和素作用实现自制液态氟碳脂质微球与损伤后心肌细胞的靶向结合。先体外培养大鼠原代心肌细胞,将培养出的心肌细胞分为TNF-α处理组和非处理组,TNF-α处理组的心肌细胞给予200ng/ml TNF-α溶液刺激。6h后两组均先后加入等量的生物素化的抗大鼠细胞间黏附分子-1(ICAM-1)单克隆抗体、链酶亲和素和自制生物素化液态氟碳脂质微球,然后用Hoechst液对两组心肌细胞核染色,最后在荧光显微镜下观察。结果显示TNF-α处理组染成蓝色的心肌细胞核周围可见大量绿色荧光微球,但非处理组蓝色的心肌细胞核周围有极少许绿色荧光微球。研究表明心肌细胞在TNF-α刺激下可产生ICAM-1炎症细胞因子,液态氟碳脂质微球联合ICAM-1单克隆抗体可通过生物素-亲和素作用实现与体外培养的大鼠损伤心肌细胞靶向连接。

The present study was to investigate in vitro the rat cardiomyocyte injury with targeting of home-made perfluorocarbon lipid particles with avidin-biotin interaction. Neonatal rat cardiomyocytes were cultured in vitro and divided into two groups： TNF-α activated group and non-activated group. Those in the TNF-α activated group were exposed to 200ng/ml TNF-α solution for 6 hours and then cardiomyocytes in both groups were pretargeted with bioti- nylated ICAM-1 monoclonal antibodies, and were exposed to streptavidin, and then to home-made green fluorescently-labeled biotinylated perfluorocarbon lipid particles. Cardiomyocytes nucleus stained with Hoechst. The results were detected with fluorescence microscope, As a result, in TNF-α activated group, around blue fluorescent cardiomyocytes nucleus, a great amount of green fluorescent particles were found, while there were few green fluorescent particles in non-TNF activated group. It has been shown that ICAM-1 is expressed in the surface of cardiomyocytes when they are stimulated by TNF-α. Perfluorocarbon lipid particles associated with ICAM-1 monoclonal antibodies can be targeted to injured cardiomyocytes by avidin-biotin interaction.