Baitouweng Tang (白头翁汤) alleviates dextran sulfate sodium-induced ulcerative colitis in mice: a network pharmacology combined with experimental study

doi:10.19852/j.cnki.jtcm.2026.01.011

Abstract

Abstract:

OBJECTIVE: To explore the mechanism of Baitouweng Tang (白头翁汤, Pulsatilla decoction, PD) alleviates dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice by integrating network pharmacology prediction with experimental validation, focusing on the modulation of inflammatory signaling.

METHODS: A chronic UC model was induced in C57BL/6 mice by cyclical administration of DSS. Mice were treated with either a low (15 mL/kg) or high (30 mL/kg) dose of PD. Disease severity was assessed clinically and via histopathology. Serum levels of inflammatory cytokines were quantified. A network pharmacology approach was employed to predict the core targets and pathways of PD against UC. Key predictions concerning the toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-κB) pathway were subsequently verified in colonic tissue using quantitative polymerase chain reaction and Western blotting.

RESULTS: PD treatment significantly ameliorated DSS-induced UC symptoms, including reducing disease activity, preventing colon shortening, and improving histological architecture. PD effectively rebalanced the systemic inflammatory milieu by decreasing pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and elevating anti-inflammatory cytokines interleukin-10 (IL-10). Network pharmacology analysis identified the TLR4/NF-κB signaling pathway as a central target. Experimental validation confirmed that PD markedly suppressed the upregulation of both TLR4 and NF-κB at the transcriptional and protein levels in the inflamed colon.

CONCLUSION: PD demonstrates protective effects against experimental UC. Its mechanism is associated with the inhibition of the TLR4/NF-κB signaling pathway and the subsequent attenuation of inflammatory responses. This study provides a modern pharmacological basis for the classical application of PD in treating heat-toxin related intestinal disorders, bridging traditional use and mechanistic understanding.

Key words: colitis, ulcerative, toll-like receptor 4, NF-kappa B, network pharmacology, Pulsatilla decoction

SUN Yan, ZHOU Haozheng, FENG Jianhui, LIN Zikai, HE Jie, WANG Zhenhua, GUO Yuting, WEN Shaohong, LI Gang. Baitouweng Tang (白头翁汤) alleviates dextran sulfate sodium-induced ulcerative colitis in mice: a network pharmacology combined with experimental study[J]. Journal of Traditional Chinese Medicine, 2026, 46(1): 119-126.

Figures/Tables 4

Figure 1 Intervention extent of PD on UC and histopathological changes in colonic tissue A: DAI scores; B: macroscopic colon appearance; C: measured colon length in the indicated groups; D: changes in body weight in the indicated groups; E: colon tissue HE staining; B1, E1, E6: control group; B2, E2, E7: model group; B3, E3, E8: mesalazine group; B4, E4, E9: PD low group; B5, E5, E10: PD high group; E1; E2; E3; E4; E5: × 100; E6; E7; E8; E9; E10: ×400. Control group: a normal control group provided with a normal diet and water; model group: a model group subjected to DSS modeling, with alternating saline and 2% DSS administration; mesalazine group: a mesalazine group mesalazine (300 mg/kg) was used to gavage mice when drinking 2% DSS; PD low group: a low-dose PD group in which mice treated with DSS were gavaged with PD (15 mL/kg); PD high group: a high-dose PD group in which in which mice treated with DSS were gavaged with PD (30 mL/kg). PD: Baitouweng Tang (Pulsatilla decoction); UC: ulcerative colitis; DAI: disease activity index; HE: hematoxylin-eosin; DSS: dextran sulfate sodium. Data were presented as mean ± standard error of the mean (n = 3). The t test was used for measurement data; one-way analysis of variance was used for multiple comparisons. Compared with the control group, aP < 0.001; compared with the model group, bP < 0.01.

Figure 2 Effect of PD on the levels of pro-inflammatory and anti-inflammatory cytokines in the serum of DSS-induced UC model mice A: IL-6. B: TNF-α. C: TGF-β. D: IL-4. E: IL-10. Control group: a normal control group provided with a normal diet and water; model group: a model group subjected to DSS modeling, with alternating saline and 2% DSS administration; mesalazine group: a mesalazine group mesalazine (300 mg/kg) was used to gavage mice when drinking 2% DSS; PD low group: a low-dose PD group in which mice treated with DSS were gavaged with PD (15 mL/kg); PD high group: a high-dose PD group in which in which mice treated with DSS were gavaged with PD (30 mL/kg). PD: Baitouweng Tang (Pulsatilla decoction); UC: ulcerative colitis; DSS: dextran sulfate sodium; IL: interleukin; TNF-α: tumor necrosis factor-alpha; TGF-β: transforming growth factor-beta. Data were presented as mean ± standard error of the mean (n = 3). The t test was used for measurement data; one-way analysis of variance was used for multiple comparisons. Compared with the control group, aP < 0.001; compared with the model group, bP < 0.001, cP < 0.01.

Figure 3 Network pharmacology analysis A: intersecting targets; B: core targets in the PPI network; C: KEGG analysis; D: associated pathways. PD: Baitouweng Tang (Pulsatilla decoction); UC: ulcerative colitis; PPI network: protein-protein interaction networks; GO: gene ontology; KEGG: kyoto encyclopedia of genes and genomes; TLR4/ NF-κB: toll-like receptor 4/ nuclear factor-kappa B; PI3K: phosphatidylinositol 3 kinase; Akt: protein kinase B; MAPK: mitogen-activated protein kinase. The abbreviations and their full names appeared in Figure 3B are presented in supplementary Table 2.

Figure 4 Effect of PD on mRNA and protein levels in the colon A: effect of PD on mRNA levels in the colon; A1: TLR4 mRNA; A2: NF-κb mRNA; B: effect of PD on protein levels in the colon; B1: TLR4 protein; B2: NF-κb protein; B3: Western blot images. Control group (1 and 2): a normal control group provided with a normal diet and water; model group (3 and 4): a model group subjected to DSS modeling, with alternating saline and 2% DSS administration;mesalazine group (5 and 6): a mesalazine group mesalazine (300 mg/kg) was used to gavage mice when drinking 2% DSS; PD low group (7 and 8): a low-dose PD group in which mice treated with DSS were gavaged with PD (15 mL/kg); PD high group (9and 10):a high-dose PD group in which in which mice treated with DSS were gavaged with PD (30 mL/kg). PD: Baitouweng Tang (Pulsatilla decoction); TLR4: toll-like receptor 4; NF-κb: nuclear factor-kappa B; DSS: dextran sulfate sodium. Data were presented as mean ± standard error of the mean (n = 3). The t test was used for measurement data; one-way analysis of variance was used for multiple comparisons. Compared with the model group, aP < 0.01; bP < 0.05; dP < 0.001; compared with the control group, cP < 0.01; eP < 0.05.

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