Cyclophosphamide (CTX), used in cancer chemotherapy, a high dose of which would cause immunosuppressive effect and intestinal mucosa damage. American ginseng (Panax quinquefolius L.) has a long history of functional food use for immunological disorder, colitis, cancer, and so on. This study aimed to illustrate the underlying mechanism of American ginseng’s immunomodulatory effect in CTX-induced mice. In this study, all groups of American ginseng (American ginseng polysaccharide [AGP], American ginseng ginsenoside [AGG], co-treated with American ginseng polysaccharide and ginsenoside [AGP_AGG]) have relieve the immune disorder by reversing the lymphocyte subsets ratio in spleen and peripheral blood, as well as stimulating CD4⁺T cells and IgA-secreting cells in small intestine. These three treatment groups, especially AGP_AGG co-treated group recovered the intestine morphology that up-regulated villus height (VH)/crypt depth (CD) ratio, areas of mucins expression, quantity of goblet cells, and expression of tight junction proteins (ZO-1, occludin). Importantly, the microbiome-metabolomics analysis was applied in this study to illustrate the possible immuno-modulating mechanism. The synergistic effect of polysaccharides and ginsenosides (AGP_AGG group) restored the gut microbiota composition and increased various beneficial mucosa-associated bacterial taxa Clostridiales, Bifidobacterium, and Lachnospiraceae, while decreased harmful bacteria Escherichia-Shigella and Peptococcaceae. Also, AGP_AGG group altered various fecal metabolites such as uric acid, xanthurenic acid, acylcarnitine, 9,10-DHOME, 13-HDoHE, LysoPE15:0, LysoPC 16:0, LysoPI 18:0, and so on, that associated with immunometabolism or protective effect of gut barrier. These results suggest AG, particularly co-treated of polysaccharide and ginsenoside may be used as immunostimulants targeting microbiome-metabolomics axis to prevent CTX-induced side effects in cancer patients.

Background:Viral pneumonia is one of the most serious respiratory diseases, and multicomponent traditional Chinese medicines have been applied in the management of infected patients. As a representative TCM, HouYanQing (HYQ) oral liquid shows antiviral activity. However, the unclear mechanisms, as well as the ambiguous clinical effects, limit widespread application of this treatment. Therefore, in this study, a proteomics-based approach was utilized to precisely investigate its efficacy.Methods:Based on the efficacy evaluation of HYQ in a mouse model of pneumonia caused by influenza A virus (H1N1) and the subsequent proteomics analysis, specific signatures regulated by HYQ treatment of viral pneumonia were identified.Results:Experimental verifications indicate that HYQ may show distinctive effects in viral pneumonia patients, such as elevated galectin-3-binding protein and glutathione peroxidase 3 levels.Conclusion:This study provides a precise investigation of the efficacy of a multicomponent drug against viral pneumonia and offers a promising alternative for personalized management of viral pneumonia.

Background: Yiqi Zengmian (YQZM) functions as an immunopotentiator by enhancing both cellular and humoral immunity. However, its pharmacodynamic active constituents, particularly those absorbed into the bloodstream, and mechanism of action remain unclear. This study aimed to investigate the immunopotentiating effects and mechanisms of YQZM in mice immunized with the BBIBP-CorV (Beijing Bio-Institute of Biological Products Coronavirus Vaccine). Methods: Serum pharmacochemistry and ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) were employed to identify bioavailable components of YQZM. The mice received the BBIBP-CorV twice on days 1 and 14, while YQZM was orally administered for 28 days. Neutralization assays and ELISA quantified antigen-specific antibodies (abs), flow cytometry (FC) and intracellular cytokine staining (ICS) were used to assess immune cell populations and their cytokines, and an enzyme-linked immunospot assay (ELISpot) quantified memory T and B cells (MBs and MTs). To identify underlying mechanisms, network pharmacology, RNA sequencing (RNA-Seq), molecular docking, Western blotting (WB), and quantitative reverse transcription PCR (RT-qPCR) were performed. Results: YQZM significantly enhanced antigen-specific antibody titers, immune cell proportions, cytokine levels, and memory lymphocyte functions. UPLC-MS/MS analysis identified 31 bioactive compounds in YQZM. KEGG enrichment analysis based on RNA-Seq and network pharmacology implicated the TLR-JAK-STAT signaling pathway in YQZM’s immune-enhancing effects. WB and RT-PCR validated that YQZM upregulated the expression of critical nodes in the TLR-JAK-STAT signaling pathway. Furthermore, molecular docking indicated that YQZM’s primary active components exhibited strong binding affinity for critical proteins. Conclusions: YQZM effectively enhances vaccine-induced innate and adaptive immunity via a multi-component, multi-target mechanism, among which the TLR-JAK-STAT signaling pathway is a validated molecular target.

Background: Dangguibuxue decoction (DD), containing Angelica sinensis (Oliv.) Diels (AS) and Astragalus membranaceus (Fisch.) Bge. (AM) (1:5), is a well-known traditional Chinese medicine (TCM) used for strengthening qi and nourishing the blood. DD has shown therapeutic effects in nephropathy patients. However, the underlying mechanisms based on the traditional efficacy are still not fully elucidated. Methods: The chemical constituents in DD were identified using UPLC-MS/MS. Network pharmacology analysis was applied to predict the potential target genes and associated signaling pathways. A renal fibrosis mouse model was induced by the intraperitoneal injection of aristolochic acid I (AA I) at 3.0 mg/kg. Mice were treated with AM, AS, and DD at two dosages by oral gavage for 30 days. Body weights, serum biochemistry, hematology, and histopathology observations were assessed. The key targets predicted were validated using qRT-PCR and Western blotting. The active constituents were screened by molecular docking, and their anti-fibrotic effects were evaluated through in vitro assays. Results: DD effectively improved renal functions and alleviated AA I-induced renal fibrosis. DD alleviated anemia and upregulated the expression of Erythropoietin (EPO). Network pharmacology analysis indicated the involvement of signaling pathways, including the PI3K/Akt, hypoxia-inducible factor-1α (HIF-1α) and transforming growth factor-β (TGF-β) signaling pathways. Experimental validation further demonstrated that DD reduced the protein expression of HIF-1α, collagen I, and TGF-β, and the ratios of phosphorylated Smad2/3 to total Smad2/3. Molecular docking and in vitro assays suggested that rutin may be a potential bioactive compound in DD. Conclusions: This research indicated that DD ameliorated AA I-induced renal fibrosis in mice, which may be associated with the modulation of HIF-1α and TGF-β/Smad signaling pathways. Rutin may be a potential bioactive compound in DD with anti-fibrotic activity, but further studies are still needed to clarify the content of rutin in DD, the amount of its exposure in the body, and its contribution to the effects of DD.

Shenqi Citian Granules (SQCT), a traditional Chinese medicine compound formula derived from Tibetan medical empirical knowledge, is composed of 15 medicinal materials including the Tibetan herb Manjing (Brassica rapa L.). It is used to treat acute mountain sickness (AMS). To scientifically evaluate the therapeutic efficacy of SQCT for AMS treatment and explore its underlying mechanisms, thereby bridging traditional ethnopharmacological knowledge with contemporary biomedical research. The SQCT bioactive constituents were identified via serum pharmacochemistry and ultra-performance liquid chromatography-tandem mass spectrometry. An AMS rat model was established using a hypobaric hypoxia chamber simulating a 7000-m altitude. The primary outcome measures included loaded swimming time and blood gas parameters and biochemical indices. The blood lactate concentrations were measured using a biochemical assay kit. The inflammatory cytokine and hypoxia-inducible factor (HIF) concentrations in lung tissue were detected using ELISA kits. Morphological changes in the lung and brain tissues were observed using H&E staining. Network pharmacology, lung tissue proteomics, and Western blot analyses were conducted to explore the potential mechanisms. UPLC-MS/MS analysis identified 36 bioactive SQCT constituents. SQCT significantly increased the loaded swimming time of AMS rats, decreased blood lactate concentrations, improved blood gas parameters, and reduced blood biochemical indices including AST concentration. It also reduced inflammatory cytokine and HIF concentrations in the lung tissue and ameliorated inflammatory and edematous lesions in both lung and brain tissues. The network pharmacology and proteomics results implicated the HIF-1 and necroptosis signaling pathways in the therapeutic effects of SQCT on AMS. Western blot experiments confirmed downregulated expressions of key nodes in both HIF-1 and necroptosis signaling pathways by SQCT. SQCT effectively treats AMS via multi-component, multi-target mechanisms, and the HIF-1 and necroptosis signaling pathways are key targets.

Honglanqi (HLQ), a distinctive commercial specification of Astragali Radix (AR) characterized by rhytidome and interxylary cork, remains undervalued despite originating from premium long-cultivated material. The ethyl acetate fraction (HLQE) showed the greatest anti-inflammatory activity among the polarity-based fractions of an HLQ ethanol extract. We applied an integrated ultra-high performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC–QTOF-MS/MS) and feature-based molecular networking (FBMN) strategy to systematically profile the chemical composition of HLQE. A total of 102 metabolites were annotated across seven compound classes, of which 18 were predicted as potential new compounds, and 37 were reported in AR for the first time. Three undescribed compounds, astragalinin A, astragalinin B, and astragquinone, were isolated and characterized, together with 19 known compounds. In LPS-stimulated RAW264.7 macrophages, astragalinin A exhibited potent inhibition of TNF-α and IL-6 (IC₅₀ < 10 μM). Astragquinone showed significant TNF-α inhibition with a favourable safety profile. Stereochemical configuration critically influenced anti-inflammatory potency, as demonstrated by the enantiomeric pair. Molecular docking predicted favourable binding orientations toward TNF-α and IL-6 in silico. These findings provide a comprehensive phytochemical and bioactivity profile of HLQ, supporting its value-added utilization as an underutilized AR resource.