In order to explore the immediate effect of polysaccharides and macrophages, polysaccharides from masson pine pollen (PPM60) were labeled with fluorescein isothiocyanate (FITC) by using a chemical-derived method, and the reactant was named PPM60-Tyr-FITC. Direct interaction of PPM60-Tyr-FITC and RAW264.7 macrophages could be detected by flow cytometer (FCM), and this interaction could be inhibited by Pitstop 2 (clathrin inhibitor) and TAK-242 (Toll-like receptor 4 inhibitor). The results of confocal laser scanning microscopy (CLSM) also revealed that there was a co-localization phenomenon between PPM60-Tyr-FITC and RAW264.7 macrophage receptors, and it could be suppressed by Pitstop 2 and TAK-242. It was confirmed that PPM60 enters into RAW264.7 macrophages mainly through endocytosis, rather than the phagocytosis, and TLR4 played a mediating role.
Keywords: PPM60-Tyr-FITC; RAW264.7 macrophages; polysaccharides.