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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。
Using biotin-avidin to effect the self-made liquid fluorocarbon lipid microspheres and the targeted binding of injured cardiomyocytes. Primary rat cardiomyocytes were cultured in vitro. The cultured cardiomyocytes were divided into TNF-α-treated group and untreated group. The myocardial cells treated with TNF-α were stimulated with 200ng / ml TNF-α. After 6h, the same amount of biotinylated monoclonal antibody against ICAM-1 (ICAM-1), streptavidin and self-made biotinylated liquid fluorocarbon lipid microspheres were successively added to the two groups, Then two groups of cardiac myocytes were stained with Hoechst solution, and finally observed under a fluorescence microscope. The results showed that a large number of green fluorescent microspheres were observed in the nuclei of myocardial cells stained blue in TNF-α-treated group, but there were very few green fluorescent microspheres in the non-treated group. Studies have shown that myocardial cells can produce ICAM-1 inflammatory cytokines stimulated by TNF-α, liquid fluorocarbon lipid microspheres combined with ICAM-1 monoclonal antibody can be achieved by biotin-avidin and rat cultured in vitro injury Cardiomyocyte-targeted ligation.