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Background <p><i>Trapa acornis</i> Nakano, also known as “Nan-hu-ling” (NHL), is an important agricultural crop whose fruits are commonly consumed as food, while its stems are usually discarded as agricultural by-products.</p> Methods and results <p>In the current investigation, a homogeneous polysaccharide (NHLP) with a weight-average molecular weight (Mw) of 62.0&#xa0;kDa was isolated from the stem of NHL. Monosaccharide composition analysis revealed that NHLP was primarily composed of Gal (24.4%), Glc (18.9%), GalA (16.8%), Xyl (12.1%), and Rha (7.7%). Methylation analysis indicated that NHLP mainly contained residues of 4-Gal<i>p</i>A, 4-Glc<i>p</i>, 4-Gal<i>p</i>, 4-Xyl<i>p</i>, 3,6-Gal<i>p</i>, t-Ara<i>f</i>, t-Gal<i>p</i>, and t-Rha<i>p</i>. The linkage patterns of these residues were structurally resolved using one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy. In vitro assays demonstrated that NHLP stimulated RAW264.7 cells to produce nitric oxide (NO) in a dose-dependent manner, promoted the release of proinflammatory cytokines including interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), and upregulated transcriptional profiles of genes involved in inflammatory responses (IL-6, IL-1β, TNF-α, COX-2, iNOS, and IκBα) and proteins (iNOS, COX-2, IL-1β, and TNF-α).</p> Conclusions <p>This study clarified the proposed structure of the polysaccharide derived from <i>T. acornis</i> stem and its macrophage-activating effects, providing supportive evidence for its potential application in immune activation.</p> Graphical abstract <p></p>

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Isolation, structural characterization, and immunostimulatory activity of a polysaccharide from the stem of Trapa acornis Nakano on RAW264.7 macrophages

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年3月27日

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  • Bin Zhang,
  • Xueping Zhang,
  • Lei Liu,
  • Xin Zhong,
  • Yiting Lyu,
  • Ke Xu,
  • Ran Zhang,
  • Xiaowen Shi

摘要

Background

Trapa acornis Nakano, also known as “Nan-hu-ling” (NHL), is an important agricultural crop whose fruits are commonly consumed as food, while its stems are usually discarded as agricultural by-products.

Methods and results

In the current investigation, a homogeneous polysaccharide (NHLP) with a weight-average molecular weight (Mw) of 62.0 kDa was isolated from the stem of NHL. Monosaccharide composition analysis revealed that NHLP was primarily composed of Gal (24.4%), Glc (18.9%), GalA (16.8%), Xyl (12.1%), and Rha (7.7%). Methylation analysis indicated that NHLP mainly contained residues of 4-GalpA, 4-Glcp, 4-Galp, 4-Xylp, 3,6-Galp, t-Araf, t-Galp, and t-Rhap. The linkage patterns of these residues were structurally resolved using one- and two-dimensional nuclear magnetic resonance (NMR) spectroscopy. In vitro assays demonstrated that NHLP stimulated RAW264.7 cells to produce nitric oxide (NO) in a dose-dependent manner, promoted the release of proinflammatory cytokines including interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), and upregulated transcriptional profiles of genes involved in inflammatory responses (IL-6, IL-1β, TNF-α, COX-2, iNOS, and IκBα) and proteins (iNOS, COX-2, IL-1β, and TNF-α).

Conclusions

This study clarified the proposed structure of the polysaccharide derived from T. acornis stem and its macrophage-activating effects, providing supportive evidence for its potential application in immune activation.

Graphical abstract