Structural Characterization and Evaluation of Anti-ulcerative Colitis Activity of Homogeneous Polysaccharide from Astragali Radix-Angelicae Sinensis Radix Herb Pair
10.13422/j.cnki.syfjx.20251766
- VernacularTitle:一种芪归均一多糖的结构表征及抗溃疡性结肠炎作用评价
- Author:
Wenjuan LIU
1
;
Shanbo MA
2
;
Ying BU
3
;
Tao MA
4
;
Xiaopeng SHI
2
;
Yuping TANG
1
Author Information
1. College of Pharmacy，Key Laboratory of Traditional Chinese Medicine（TCM） Compatibility of Shaanxi Administration of TCM，Shaanxi University of Chinese Medicine，Xianyang 712046，China
2. The First Affiliated Hospital of Air Force Medical University，Xi'an 710032，China
3. Third Affiliated Hospital of Naval Medical University，Shanghai 200433，China
4. The 968th Hospital of Joint Logistics Support Force of the Chinese People's Liberation Army，Jinzhou 121000，China
- Publication Type:Journal Article
- Keywords:
Astragali Radix-Angelicae Sinensis Radix herb pair;
polysaccharides;
high performance liquid chromatography（HPLC）;
gas chromatography-mass spectrometry（GC-MS）;
nuclear magnetic resonance（NMR）;
immunoregulation;
ulcerative colitis（UC）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(20):204-213
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the immunomodulatory effect of polysaccharides from Astragali Radix-Angelicae Sinensis Radix herb pair（Qi-gui polysaccharides） on lipopolysaccharide（LPS）-induced RAW264.7 macrophages and to characterize the structure of the active component Qi-gui homogeneous polysaccharide（AAPS-4a）， and evaluate its protective effect on ulcerative colitis（UC）. MethodsThe effects of six Qi-gui polysaccharides（0.01-100 mg·L-1） on the proliferation of RAW264.7 cells were assessed by cell proliferation and activity assay（CCK-8）， and enzyme-linked immunosorbent assay（ELISA） was used to investigate the effects of the six polysaccharides（3， 10 mg·L-1） on the secretion levels of tumor necrosis factor（TNF）-α， interferon（IFN）-β， and nitric oxide（NO） in LPS-induced RAW264.7 cells. After screening for active polysaccharides， high-performance size-exclusion chromatography（HPSEC） was used to determine its homogeneity and relative molecular weight， then its characteristic functional groups were identified by Fourier transform infrared spectroscopy（FT-IR）， monosaccharide composition was analyzed by high performance liquid chromatography（HPLC）， methylation analysis combined with gas chromatography-mass spectrometry（GC-MS） was performed to determine the types and linkage modes of sugar residues， and one- and two-dimensional nuclear magnetic resonance（NMR） were used to identify the sugar residue composition and configuration of the active polysaccharide. Finally， experimental animals were divided into the normal group， model group， AAPS-4a low-dose group（50 mg·kg-1）， AAPS-4a high-dose group（100 mg·kg-1）， and sulfasalazine（SASP） group （75 mg·kg-1）. Except for the normal group， the acute UC mouse model was induced using 3.5% dextran sulfate sodium salt（DSS）. Each treatment group was administered the corresponding dose via oral gavage for 7 days， and changes in body weight were recorded. After treatment， the spleen index and disease activity index（DAI） score were calculated， TNF-α and interleukin-6（IL-6） levels in the serum were detected by ELISA， and histopathological changes in colon tissue were observed by hematoxylin-eosin（HE） staining. ResultsAt the cellular level， AAPS-4a exhibited a dose-dependent inhibition of LPS-induced increases in TNF-α， IFN-β， and NO levels（P<0.01）. Structural characterization of AAPS-4a revealed that it was a homogeneous polysaccharide with a relative molecular weight of 7.6×103 Da， consisting of mannose（Man）， glucose（Glc）， and galactose（Gal） in a molar ratio of 1.3∶23.9∶1.0. It was primarily composed of five sugar residues of 1，6-α-D-Glcp， T-α-D-Glcp， 1，3-β-D-Galp， 1，4-α-D-Manp， and 1，2-α-D-Galp. In vivo experiments showed that compared with the normal group， the model group demonstrated markedly increased DAI score and spleen index， significantly reduced colon length， and significantly elevated levels of TNF-α and IL-6（P<0.01）. Compared with the model group， the AAPS-4a high-dose group significantly reduced the DAI score and spleen index， as well as TNF-α and IL-6 levels， and improved colonic atrophy（P<0.05， P<0.01）. Pathological observations showed that AAPS-4a significantly inhibited inflammatory cell infiltration in colon tissue and alleviated pathological damage. ConclusionAAPS-4a， a neutral homogeneous polysaccharide composed of 1，6-α-D-Glcp， T-α-D-Glcp， 1，3-β-D-Galp， 1，4-α-D-Manp and 1，2-α-D-Galp， is identified as a key bioactive component contributing to the anti-UC effect of the Qi-gui herb pair. Its immunoregulatory and anti-UC properties suggest its potential as a therapeutic agent for UC.
