The Chemical Profiling and Immunological Activity of Polysaccharides from the Rhizome of Imperata cylindrica Using Hot Water Extraction

To investigate the immunomodulatory activity of polysaccharides derived from the rhizome of Imperata cylindrica, polysaccharides (IRPs-H) were extracted using hot water extraction and further purified via DEAE-52 ion-exchange chromatography, yielding three fractions: IRPs-H1, IRPs-H2, and IRPs-H3. The structural features of these fractions were characterized by Fourier-transform infrared spectroscopy (FT-IR), high-performance gel permeation chromatography (HPGPC), atomic force microscopy (AFM), and thermogravimetric analysis (TGA). Their immunological activities were evaluated in vitro. All three fractions were identified as neutral pyranose-type polysaccharides, primarily composed of glucose and xylose, exhibiting good thermal stability and lacking long-chain structures. In vitro assays using RAW264.7 macrophages demonstrated that these polysaccharides promoted cell proliferation (50–800 μg/mL), enhanced phagocytic activity, and induced morphological changes characteristic of macrophage activation, including irregular shapes and pseudopod formation. ELISA and flow cytometry analyses revealed dose-dependent increases in nitric oxide (NO), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and reactive oxygen species (ROS) levels. Notably, the IRPs-H3 fraction stimulated TNF-α and IL-6 production to levels of 438.02 ± 14.14 pg/mL and 30.13 ± 1.27 pg/mL, respectively, which were comparable to those induced by lipopolysaccharide (LPS), the positive control (460.83 ± 16.10 pg/mL and 31.87 ± 1.72 pg/mL, respectively). These results suggest that polysaccharides extracted from the rhizome of Imperata cylindrica possess significant immunostimulatory properties and hold potential for development as functional food ingredients or immune-enhancing agents.