矢车菊素-3-O-葡萄糖苷进入巨噬细胞的荧光检测  

Fluorescence Detection of the Entry of Cyanidin-3-O-Glucoside into Macrophages

作　　者：张英慧[1] 王丙云[2] 黄剑波[1] 计慧琴[2] 马艳玲[1] 钟希琼[1] 董华强[1] 

机构地区：[1]佛山科学技术学院北院食品安全系,广东南海528231 [2]佛山科学技术学院北院动物医学系,广东南海528231

出　　处：《食品科学》2016年第9期198-202,共5页Food Science

基　　金：国家自然科学基金项目面上项目(31071537);广东省科技计划项目(2011A030100018);佛山市科技创新专项资金项目(2014GA000425)

摘　　要：利用荧光技术探索矢车菊素-3-O-葡萄糖苷(cyanidin 3-O-glucoside,C3G)在体外培养的巨噬细胞中的分布。首先采用荧光分光光度法扫描C3G的特异激发波长和发射波长,再利用激光共聚焦技术探讨C3G进入小鼠和人巨噬细胞的过程以及C3G在细胞中的定位,并分析C3G孵育时间对细胞内荧光强度变化的影响。结果表明:在488 nm和520 nm的激发波长下,C3G孵育15 min的细胞质内开始呈现绿色和红色荧光,并且随着孵育时间的变化,细胞内荧光强度逐渐增强,其中孵育60 min可观察到荧光布满细胞核,其荧光强度是孵育前的6.45倍。研究表明采用特异波长的激光共聚焦断层扫描技术可示踪到花色苷在干预细胞内的分布,结果显示花色苷C3G可快速穿过巨噬细胞的细胞膜和核膜,直达细胞核。The distribution of cyanidin-3-O-glucoside（C3G） in cultured mouse and human macrophages was detected with fluorescence technique. First, the specific excitation wavelength and emission wavelength of C3 G were scanned with fluorescence spectrophotometric assay. Then, a confocal laser scanning microscopy was utilized to investigate the entry process of C3 G into cultured mouse macrophages and human macrophage THP-1 cells, and to trace the location of C3 G as well as the change in fluorescence intensity in the cells during incubation. The results showed that green and red fluorescence in macrophages after 15 min C3 G incubation could be detected with light excitation at 488 nm and 520 nm, respectively. The fluorescence signals were enhanced in the cells as the incubation time was prolonged. The special fluorescence appeared in nuclei when the cells were treated with C3 G for 60 min, and the average fluorescence intensity was enhanced by 6.45 folds when C3 G incubation time was increased up to 60 min. These results indicated that the distribution of C3 G in cells during incubation could be traced with laser scanning confocal microscopy assay. It was concluded that C3 G could pass rapidly through the cell membrane and nuclear membrane of macrophages, and entered directly the cell nucleus.

关 键 词：矢车菊素-3-O-葡萄糖苷 巨噬细胞 激光共聚焦显微技术 荧光分光光度法 

分 类 号：TS201.4[轻工技术与工程—食品科学]