Treatment with Biaoxu Ganmao Granules ameliorates cyclophosphamide-induced immunosuppression in mice: Paeoniflorin-associated modulation of the TLR4/NF-κB pathway.

Nonpolysaccharide fraction of Lonicerae japonicae Flos attenuates cyclophosphamide-induced immunosuppression associated with modulation of the Keap1/Nrf2/HO-1/GPX4 signalling pathway

Background Cyclophosphamide (CTX)-induced immunosuppression is closely associated with oxidative stress and ferroptosis, but effective therapeutic interventions remain limited. This study investigated the protective effects of the non-polysaccharide fraction of Lonicerae japonicae Flos (LJFE) against CTX-induced immunosuppression and explored its underlying molecular mechanism. Methods LJFE was prepared by water extraction followed by ethanol precipitation, and its chemical profile was characterized using UPLC-MS/MS. Mice with CTX-induced immunosuppression were administered LJFE at doses of 100, 200, and 400 mg/kg for 10 consecutive days. Immune organ indices, serum IgE and IgM levels, oxidative stress markers (SOD, GSH, MDA), and the TBX21/GATA3 ratio were measured. The Keap1/Nrf2/HO-1/GPX4 signalling pathway was investigated via Western blotting and immunofluorescence. Serum metabolomics and molecular docking were applied to screen key differential metabolites and predict binding interactions between characteristic components and target proteins. LPS-stimulated RAW264.7 macrophages were used to verify pathway activation. Results Seventeen chemical constituents were identified in LJFE. In vivo, LJFE markedly restored thymus and spleen indices, increased serum IgE and IgM levels by approximately 45% and 35%, respectively, elevated SOD and GSH activities, reduced MDA accumulation, and normalized the TBX21/GATA3 ratio. Mechanistically, LJFE downregulated Keap1 and upregulated Nrf2, HO-1, and GPX4 expression. Metabolomics analysis identified hydroferulic acid as a key differential biomarker involved in antioxidative regulation. Molecular docking suggested that characteristic components of LJFE exhibit favorable binding affinity toward Keap1 and Nrf2. In vitro, LJFE dose-dependently activated the Keap1/Nrf2/HO-1 signalling axis and suppressed excessive NO release in LPS-challenged macrophages. Conclusion LJFE ameliorates CTX-induced immunosuppression by modulating the Keap1/Nrf2/HO-1/GPX4 pathway, thereby alleviating oxidative stress and ferroptosis. These findings support the potential of LJFE for further development as an immune-enhancing functional food ingredient.

Siweixizangmaoru Decoction Alleviates Rheumatoid Arthritis by Enhancing Pol β-Mediated Attenuation of DNA Damage and Suppression of cGAS/STING/NF-κB/NLRP3-Associated Pyroptosis

Objective: Siweixizangmaoru decoction (SXD), a classical Tibetan prescription documented in the medical text Four Medical Tantras, has shown therapeutic activity in experimental rheumatoid arthritis (RA). However, its candidate active constituents and underlying mechanisms remain unclear. Methods: In this study, serum-absorbed constituents following SXD administration were profiled using ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS), and network pharmacology was employed to identify core targets and candidate active constituents of SXD against RA. Candidate active constituents were further screened using cell-based assays, and the selected constituents were quantified using HPLC fingerprint analysis. Both in vivo (CIA rat model) and in vitro (RAW264.7 pyroptosis model) systems were used to investigate the pharmacological effects and mechanisms of SXD. Results: A total of 11 absorbed constituents were identified in the serum of SXD-treated rats, and four preliminary candidate active constituents, including chebulagic acid, kaempferol, gentiopicroside, and berberine, were screened. The combined in vivo and in vitro results showed that SXD attenuated RA progression and reduced inflammatory cytokine levels in rat serum and cell culture supernatants. At the molecular level, SXD increased Pol β expression and reduced DNA damage markers, cytosolic dsDNA accumulation, and cGAS/STING-, NF-κB-, and NLRP3-associated signaling. Pol β knockdown partially reduced the effects of SXD. Conclusions: These findings support the partial involvement of Pol β-associated processes in the anti-arthritic effects of SXD.

18β-Glycyrrhetinic acid monoglucuronide (GAMG) alleviates radiation-induced lung injury in mice by regulating macrophage polarization through the NF-κB signaling pathway.

PURPOSE Radiation-induced lung injury (RILI) is an unforeseeable outcome of radiotherapy, and there are yet no feasible alternatives to treatment. The objective of this study is to characterize the protection of RILI by 18β-Glycyrrhetinic acid monoglucuronide (GAMG, also referred to as glycyrrhetinic acid 3-O-mono-β-D-glucuronide)3-O-mono-β-D-glucuronide) and to explore the molecular mechanisms that may be implicated. METHODS A murine model of radiation-induced lung injury (RILI) was established by administering a single 16 Gy dose of thoracic irradiation to the mice, followed by continuous intervention with GAMG. The investigation focused on assessing morphological and pathological alterations in lung tissue, quantifying inflammatory mediators, evaluating collagen deposition, and analyzing the expression of α-smooth muscle actin (α-SMA). Histopathological changes and collagen deposition were examined using Hematoxylin and Eosin (H&E) and Masson trichrome staining techniques. Cytokine concentrations in bronchoalveolar lavage fluid (BALF) were quantified via enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry (IHC) was utilized to assess markers indicative of M1-like macrophage polarization, specifically CD86 and inducible nitric oxide synthase (iNOS). Macrophage polarization status was further analyzed using flow cytometry. The activity of the NF-κB signaling pathway in lung tissue was investigated through Western blot analysis. RESULTS The administration of GAMG effectively reduced symptoms of RILI and mitigated lung tissue damage in mice. Mechanistic studies revealed that GAMG inhibited radiation-induced activation of the NF-κB signaling pathway, while suppressing the differentiation of macrophages toward the pro-inflammatory M1-like phenotype in lung tissue. Further experiments showed that GAMG regulation of macrophage polarization may be achieved by inhibiting the NF-κB pathway. CONCLUSION GAMG may decrease RILI via inhibiting NF-κB-induced inflammation and regulating macrophage polarization. This work aims to clarify the mechanism of action of GAMG, which could potentially serve as a therapeutic agent for preventing and treating lung damage caused by radiation.

Ludangshen oral liquid ameliorates cyclophosphamide-induced immunosuppression in mice: Insights from metabolomic, gut microbiota, and multi-omics correlation analyses.

Immune dysfunction elevates susceptibility to infections, diseases, and tumors. While conventional immunomodulators often cause adverse effects, plant-derived therapies offer superior stability and hold considerable research promise. Codonopsis pilosula (Franch.) Nannf., a traditional medicinal herb, replenishes blood, promotes fluid production, and tonifies qi. Ludangshen Oral Liquid (LDSOL), a preparation derived from C. pilosula, contains multiple bioactive constituents, including lobetyolin; however, its immunomodulatory effects and their associations with host metabolism and gut microbiota remain unclear. In this study, we investigated the effects of LDSOL in cyclophosphamide-induced immunosuppressed mice using pharmacodynamic evaluation, metabolomics, 16S rRNA sequencing, and multi-omics correlation analysis. Our results demonstrated that LDSOL alleviated hepatosplenomegaly and thymic atrophy, regulated blood cell counts, and elevated levels of IL-2, IL-6, TNF-α, IFN-γ, and IgG. It also restored the balance between Th17/Treg and Th1/Th2 subsets, suggesting improved immune responses. Metabolomic and 16S rRNA analyses revealed that LDSOL modulated endogenous metabolites (e.g., L-phenylalanine, serotonin) and gut bacterial communities (e.g., Lactobacillus, Ruminococcus). Integrated correlation analysis further identified potential associations among altered gut microbial taxa, LDSOL-responsive metabolites, and immune-related parameters. Collectively, these findings indicate that LDSOL ameliorates cyclophosphamide-induced immunosuppression in mice and suggest that metabolic remodeling and gut microbiota alterations may be associated with its immunomodulatory effects.

Pentoxifylline targets TLR4/MyD88/NF-κB signaling to ameliorate neuroinflammation and metabolic dysfunction in a rat model of chronic hypoperfusion-induced vascular cognitive impairment

Cistanche deserticola Polysaccharide Ameliorates Cyclophosphamide-Induced Splenic Immunosuppression in Mice: Integrated Transcriptomic and Proteomic Analyses of Immune Modulation

Cistanche deserticola polysaccharide (CDP) exhibits pleiotropic bioactivities, yet its splenic-protective profile remains incompletely defined. Here, male ICR mice were challenged with cyclophosphamide (CTX) to establish an immunosuppressed model and concurrently treated with CDP. By integrating functional assays, splenic histopathology, and transcriptomic and proteomic analyses, we show that CDP dose-dependently restores splenic mass, rescues white-pulp atrophy, and suppresses megakaryocytic hyperplasia. Functionally, CDP rebalances pro-/anti-inflammatory cytokines, scavenges splenic ROS/MDA, and potentiates GSH-Px/SOD antioxidant capacity versus CTX alone. Multi-omics convergence (3103 DEGs; 1387 DEPs) delineated a CDP-distinctive signature related to innate immune recognition, oxidative stress buffering, and protease/ion-transport modules. Notably, transcriptional enrichment of neutrophil extracellular trap (NET)-associated proxies—synergized with NOD-like receptor/IL-17 signaling—suggests a putative innate-priming mechanism warranting functional validation, rather than confirmed pathway activation. Collectively, CDP mitigates CTX-induced splenic injury primarily through coupled redox restoration and transcriptional-level immune modulation.