Search Results (431)

Jing Guan Fang (JGF) is a traditional multi-herb formula used for hyperinflammatory conditions associated with severe viral infections; however, its protein-level effects in macrophage-mediated inflammation remain incompletely characterised. In this study, we combined ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS)-based phytochemical profiling, functional anti-inflammatory assays, and discovery-level proteomic and secretome analyses to investigate the anti-inflammatory effects of JGF in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Phytochemical profiling revealed predominantly flavonoid- and phenolic-related annotated features, together with selected terpenoid- and alkaloid-related annotations. Functionally, JGF treatment was associated with reduced nitric oxide production and differential effects on pro-inflammatory cytokine release, with a clearer concentration-dependent reduction in interleukin-6 (IL-6) and a more variable tumour necrosis factor-α (TNF-α) response, without reducing cell viability. Label-free quantitative proteomics suggested that JGF treatment was associated with modulation of LPS-induced protein changes, including reduced abundance of inflammation-associated proteins such as myristoylated alanine-rich C-kinase substrate (MARCKS) and inducible nitric oxide synthase (NOS2), together with partial restoration of selected regulatory proteins such as AIMP1 and AIMP2. Secretome analysis further suggested that JGF reduced extracellular inflammatory and tissue-remodelling-related mediators, including SERPINE1/PAI-1. Exploratory pathway analysis indicated that JGF treatment was associated with changes in inflammation-related and oxidative stress-related signalling networks. Collectively, these findings provide discovery-level molecular insights into the anti-inflammatory effects of JGF in LPS-stimulated macrophages and support further targeted validation of its pharmacological activity as a candidate multi-component anti-inflammatory formula. Full article

Bioactive food ingredients that regulate the gut microbiota are promising dietary strategies for supporting systemic health, but their roles in skin photoaging remain insufficiently defined. Qiongyu Gao (QYG), a classical food–medicine homologous formula composed of Rehmanniae Radix, Panax ginseng, and Poria cocos, was evaluated as an oral functional food candidate for UV-induced skin photoaging. QYG was chemically characterized by HPLC and UPLC–QTOF–MS/MS. Young and aged mice were subjected to D-galactose plus UVA/UVB exposure and orally administered QYG, followed by skin transcriptomics, gut microbiota sequencing, serum metabolomics, and validation in UVB-injured primary dermal fibroblasts. QYG alleviated wrinkle formation, epidermal thickening, oxidative stress, inflammation, extracellular matrix degradation, collagen disorganization, and hyaluronic acid loss. Multi-omics analysis showed that QYG selectively remodeled gut microbiota, enriching Lactobacillus-, Bifidobacterium-, and Akkermansia-associated taxa, regulated serum metabolites related to energy and lipid metabolism, and enriched mitochondrial energy metabolism-related pathways in photoaged skin. In fibroblasts, QYG-containing serum restored mitochondrial membrane potential, reduced ROS accumulation and cellular senescence, and regulated AMPK/PGC-1α-associated markers. These findings support QYG as a promising food–medicine homologous functional food candidate for skin health maintenance through gut microbiota-associated systemic metabolic regulation. Full article

Background/Objectives: Water deficit during the pod filling stage severely limits the productivity of common beans (Phaseolus vulgaris). Although the effects of water scarcity have been extensively studied in leaves and roots, the contribution of xylem sap to systemic metabolic adaptation remains poorly understood. This study investigated genotype-specific metabolic changes in xylem sap exudates in response to water deficit in two common bean genotypes with contrasting growth rates. The OTI genotype has a growth cycle of 120 days, while Rosa La Bufa (RB) completes its cycle in 80 days under both well-watered and water deficit conditions. Methods: Physiological responses were evaluated, and xylem sap exudate metabolites were profiled using untargeted UPLC–ESI–QTOF–MS. Protein–metabolite interaction networks were reconstructed using STITCH v5 to identify genotype-specific metabolic organization under stress. Results: Water deficit reduced the abundance of multiple xylem metabolites, including flavonoids, isoflavones, phenolic acids, sugars, amino acids, and oxylipin-related compounds, indicating systemic metabolic contraction. OTI exhibited extensive metabolic changes characterized by enrichment of citrate, aromatic amino acids, glycolytic intermediates, flavonoid glycosides, and detoxification-associated metabolites, consistent with active carbon remobilization and oxidative stress responses. In contrast, RB accumulated isoflavones, including genistein, biochanin A, and baicalein, together with glycosylated triterpenoid saponins. Network analysis revealed that OTI developed a broad stress-responsive interactome that integrated phenylpropanoid metabolism, carbon remobilization, and detoxification pathways, whereas RB maintained a compact interactome reinforced by oxylipin-, jasmonate-, and isoflavone-associated modules. Conclusions: Xylem sap undergoes genotype-dependent metabolic adjustment under water deficit, suggesting contrasting drought adaptation strategies. Long growth cycle genotypes activate extensive metabolic plasticity, whereas reduced growth cycle genotypes rely on more specialized defense networks. These findings highlight xylem metabolomics as a valuable approach for understanding systemic drought adaptation and identifying putative metabolic biomarkers associated with drought resilience in common beans. Full article

Argemone ochroleuca and Argemone mexicana are widespread weed species known for being rich in various secondary metabolites. However, a comprehensive understanding of their chemical diversity remains limited. Insufficient information exists on metabolite variation between plant parts and the influence of solvent polarity on metabolite recovery. Therefore, this study aimed to characterize the metabolomic profiles of the shoots and root extracts of both species using solvents of varying polarity to evaluate plant part-specific metabolite distribution and solvent effects on metabolome coverage. Untargeted ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS)-based metabolomics and molecular networking were employed for metabolite analysis. Principal component analysis (PCA) did not reveal clear clustering of A. ochroleuca and A. mexicana, suggesting similarities in their metabolomic profiles. A total of 15 and 13 metabolite classes, yielding 59 and 54 metabolites, were identified in A. ochroleuca and A. mexicana, respectively, including flavonoids, terpenoids, phenolic compounds, fatty acids, and monoterpenoids. Argemone ochroleuca exhibited higher metabolite abundance, particularly in methanol and acetone extracts, and with shoots showing higher abundance than roots, with flavonoids being the dominant class. The findings show that LC-MS metabolomics, molecular networking, and careful solvent selection are effective for identifying key metabolites with potential applications in crop protection. Full article

Orchid seeds are dust-like and lack endosperm, which limits their capacity to support germination using endogenous nutrient reserves. Successful germination therefore depends on the establishment of a compatible symbiotic association with germination-promoting orchid mycorrhizal fungi (OMF). In plant-root symbioses, host-derived small molecules, such as strigolactones and flavonoids, function as early chemical signals that recruit microbial partners and stimulate their growth. However, the chemical constituents that may facilitate fungal recognition, growth, or colonization during orchid seed germination remain poorly understood. In this report, we combined metabolomics, UPLC-Q-TOF-MS/MS, molecular networking, phytochemical isolation, and bioactivity assays to characterize bioactive metabolites from the seeds of the medicinal orchid Cremastra appendiculata. Metabolomic profiling revealed abundant primary metabolites, including lipids, amino acids, organic acids, saccharides, and nucleosides. And then, nineteen secondary metabolites were isolated and identified, including two new structures. Functional assays showed that selected organic acids, saccharides, and lignanamides promoted the growth of Coprinellus disseminatus, a fungus required for seed germination, whereas lignanamides inhibited the plant pathogen Fusarium oxysporum. These findings provide the first systematic chemical and functional characterization of metabolites from C. appendiculata seeds and offer new insight into the molecular basis of symbiosis between orchids and fungi. Full article

Agathosma betulina (P.J. Bergius) Pillans is an aromatic medicinal shrub endemic to South Africa’s Cape Floristic Region, widely used in traditional medicine to treat urinary tract infections, gastrointestinal disorders, colds, and wounds. Despite its commercial importance, limited information is available on how different growing environments influence its phytochemical composition. This study profiled the secondary metabolite composition of A. betulina leaves collected from wild, open-field cultivation, and glasshouse-grown environments using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) combined with chemometric analyses. Methanolic extracts prepared from nine samples were analysed, resulting in the confident annotation of 21 secondary metabolites (MSI Level 2), comprising phenolic acids, flavanones, flavones, and flavonol glycosides. Rutin was the predominant flavonoid, with concentrations of 194.62, 185.71, and 58.82 mg CE kg⁻¹ DW in wild, cultivated, and glasshouse-grown plants, respectively. Neochlorogenic acid accumulated predominantly in cultivated plants (51.84 mg CE kg⁻¹ DW) compared with wild plants (3.31 mg CE kg⁻¹ DW), while coumaroylquinic acid (125.93 mg CE kg⁻¹ DW) and feruloylquinic acid (30.74 mg CE kg⁻¹ DW) were most abundant under glasshouse conditions. Tricin-7-O-glucoside (62.93 mg CE kg⁻¹ DW) and isorhamnetin-3-O-glucoside (31.26 mg CE kg⁻¹ DW) also accumulated significantly in cultivated plants. Partial Least Squares Discriminant Analysis (PLS-DA) score explained 92.1% of the total variation (C1 = 63.8%; C2 = 28.3%), together with hierarchical heat map clustering and VIP, clearly separating samples according to growing environment. Wild plants exhibited the greatest metabolite diversity, cultivated plants showed intermediate metabolite profiles, and glasshouse-grown plants displayed reduced flavonoid accumulation and overall chemical diversity. These findings demonstrate that growing environment strongly influences secondary metabolite accumulation in A. betulina and provide valuable information for optimizing cultivation practices, quality control, and conservation strategies for this economically important medicinal species. Full article

Lobostemon fruticosus (L.) H. Buek is an indigenous South African medicinal plant valued for its traditional therapeutic uses; however, information on its phytochemical variation and pyrrolizidine alkaloid (PA) content across geographical populations remains limited. This study investigated the secondary metabolite composition and PA accumulation in nine biological samples (three geographical locations × three biological replicates) of L. fruticosus collected from Rhodes Memorial, Doringrivier, and Grootnek Fontein in the Western Cape Province between January and March 2025 using untargeted UPLC-QTOF-MS and targeted LC-MS/MS, with values presented as mean ± standard deviation (SD). A total of 15 secondary metabolites of different classes were putatively annotated. Seven compounds, including kaempferol, jaceidin 7-rhamnoside, schizotenuin F, senburiside III, cosmosporaside B, lirioresinol A, and (−)-steganacin, are reported for the first time in L. fruticosus. Significant geographical variation (p < 0.05) was observed, with Doringrivier containing the highest relative abundances of rutin (4815.60 ± 18.96 mg CE g⁻¹ DW), rosmarinic acid (3846.89 ± 285.78 mg CE g⁻¹ DW), and rabdosiin (5142.10 ± 651.00 mg CE g⁻¹ DW). Six of the ten targeted PAs were detected, with lycopsamine N-oxide being the predominant alkaloid (3.89 mg kg⁻¹ in Grootnek Fontein). Principal component analysis explained 97.2% of the total metabolomic variation (PC1 = 72.6%; PC2 = 24.6%) and clearly separated the three geographical populations. The OPLS-DA model showed strong discrimination among populations (R²X = 0.725, R²Y = 0.998, and Q² = 0.994), supporting the presence of location-specific chemotypic differences. The observed chemotypic differences may be associated with variation in soil properties, climatic conditions, water availability, altitude, and nutrient status among the study sites. Collectively, the results provide a foundation for chemotype selection, quality control, conservation, and the safe medicinal utilization of this species. Full article

Background: Honghema is a traditional Chinese medicinal material and is also widely used by ethnic minority communities in Guizhou Province, where it is commonly known as “Reib ndad gunb” and “Uab Detdend”. The botanical origins were the whole plants of Laportea bulbifera (Siebold & Zucc.) Wedd. (L. bulbifera) and Girardinia diversifolia (Link) Friis (G. diversifolia). Due to morphological similarities, Urtica mairei H. Lév. (U. mairei) was often mixed as easily confusable species in actual market circulation. However, systematic research regarding the differences in chemical composition between the two botanical origins, as well as between them and their easily confusable species remain limited. This study aimed to comprehensively characterise the chemical constituents of L. bulbifera, G. diversifolia, and U. mairei, and to identify significantly differential chemical constituents for their discrimination. Methods: Data acquisition was performed using ultra-performance liquid chromatography–mass spectrometry (UPLC-MS). Multivariate statistical analyses combined with reference standard comparison were performed to investigate the differences in chemical profiles among L. bulbifera, G. diversifolia and U. mairei, identify the key differential constituents leading to the differences, and clarify their relative abundance information. Results: A total of 133, 77, and 59 chemical constituents were identified from L. bulbifera, G. diversifolia, and U. mairei, respectively. These compounds comprised flavonoids, organic acids, carbohydrates, organonitrogen compounds, terpenoids, coumarins, lipids, steroids, and other types of compounds. Flavonoids constituted the highest proportion of the compounds in L. bulbifera, whereas organic acids were the most abundant in G. diversifolia and U. mairei. Through multivariate statistical analysis, 32 inter-group differential components between L. bulbifera and G. diversifolia, 27 between L. bulbifera and U. mairei, and 13 between G. diversifolia and U. mairei were screened and identified. The core components contributing to these differences included Baimaside, Roxburic acid, 1-Galloyl-glucose, Protocatechuic acid 3-glucoside, and Schaftoside. Venn diagram analysis revealed 20 characteristic differential constituents of L. bulbifera, mainly represented by flavonoids; 6 characteristic differential constituents of G. diversifolia, primarily comprising terpenoids and organic acids; and 4 characteristic differential constituents of U. mairei, mainly represented by carbohydrates. Conclusion: Significant differences were observed among the chemical profiles of the three plant species. The method established in this study can effectively identify the chemical compositions of Honghema and its easily confusable species, revealing the key components responsible for the variations in their chemical profiles. This study provides a basis for elucidating the material foundation of Honghema and offers an effective analytical approach and valuable reference data for distinguishing Honghema from its easily confusable species. Full article

This study aimed to screen for optimal Traditional Chinese medicine (TCM) combinations and subsequently optimize their extraction processes for developing antibacterial dressings by preparing aqueous and hydroethanolic extracts from Herba Centellae asiaticae, Flos Calendulae, Folium Aloe, Cortex Betulae and Rhizoma Curcumae longae and evaluating their inhibitory activity against E. coli, S. aureus, C. albicans, and T-Salmonella using an agar diffusion assay. Synergistic combinations were identified through a grid screening method, with extraction processes optimized via single-factor experiments and Box–Behnken response surface design. The results demonstrated that the Folium Aloe and Cortex Betulae formulation exhibited the strongest synergistic antibacterial activity, with the inhibition zone expanding from 11.1/12.8 mm for individual components to 17 mm after combination, representing an increase of 21–36%. Single-factor and response surface optimization identified optimal process parameters as a ratio of 1.53:1, a hydroethanolic concentration of 79.5%, and an extraction time of 2.6 h, with a model R² value ranging from 0.9265 to 0.9783; the MIC values were reduced by 25–68.75%. The UPLC-QTOF-MS analysis of the optimized extract identified nine major constituents, including anthraquinones, chromones, and triterpenoids, providing the phytochemical basis for the observed bioactivities. Mechanistic studies via UV-Vis spectrophotometry and propidium iodide (PI) staining confirmed membrane disruption through nucleic acid and protein leakage. Cytocompatibility evaluation using L929 fibroblasts demonstrated >90% cell viability even at 1000 μg/mL. Additionally, the optimized extract exhibited concentration-dependent antibacterial activity, achieving a DPPH scavenging rate of 90% at 1.0 mg/mL, whereas its ABTS scavenging rate was comparable to that of ascorbic acid. The Folium Aloe and Cortex Betulae pair demonstrates dual antibacterial and antioxidant activities, providing an experimental basis for developing functional wound dressings and green antibacterial biomaterials. Full article

Natural products represent a valuable source of anticancer agents, although their mechanisms of action are often incompletely understood. In this study, we evaluated the cytotoxic effects of the ethyl acetate (EA) fraction derived from Schima superba bark. The EA fraction exhibited selective cytotoxicity against HepG2 and MCF-7 cancer cells, with minimal effects on normal HEK293 cells, and induced apoptosis as evidenced by time- and dose-dependent activation of caspase-3. Phytochemical profiling by UPLC-QTOF-MS/MS identified 18 major constituents, predominantly phenylethanoid glycosides and triterpenoids. To gain insight into the underlying mechanism, molecular docking, 200 ns molecular dynamics simulations, and MM-PBSA analyses were performed targeting poly (ADP-ribose) polymerase 1 (PARP1) and caspase-3. Among the identified compounds, cistanoside D displayed favorable binding affinity toward both targets, but formed a more stable and energetically favorable complex with PARP1 during molecular dynamics simulations, whereas its interaction with caspase-3 was comparatively weak. These findings suggest that cistanoside D may preferentially interact with PARP1, while the observed activation of caspase-3 is likely associated with downstream apoptotic processes rather than direct enzymatic modulation. This study provides an integrated evaluation of the phytochemical composition and anticancer potential of Schima superba bark extract, and identifies cistanoside D as a promising candidate for further investigation. These results contribute to a better understanding of the molecular basis underlying the bioactivity of this medicinal plant and support its potential as a source of anticancer agents. Full article

Cardiac glycosides from Periplocae Cortex are bioactive constituents, but isolating and structurally confirming their trace in vivo metabolites remains challenging because authentic standards are often unavailable. We developed a steroid-core fragment-guided strategy based on the diagnostic ion at m/z 355.2270 that integrates metabolite screening, targeted purification, and structural characterization. Periplocin-related analytes were detected in rat urine and plasma after oral periplocin administration using HPLC-QQQ-MS/MS and UPLC-Q-TOF-MS/MS. Guided by the diagnostic ion, 13 L of rat urine was fractionated using macroporous resin, Sephadex LH-20, ODS chromatography, and preparative HPLC. Two trace metabolites were isolated and identified as gomphogenin (M1, 4 mg) and 17α-asclepioside (M2, 8 mg) by HR-MS and NMR spectroscopy. Periplocin, periplogenin, and periplocymarin were confirmed using reference standards. Structural comparison supported a putative transformation pathway involving deglycosylation and steroid-core modification. In a hypoxia/reoxygenation-injured H9c2 cell model, M1 and M2 increased cell viability at 50 μM, supporting further evaluation using complementary cellular endpoints. This diagnostic-ion-guided workflow enables the targeted isolation of trace cardenolide metabolites from biological matrices and provides reference compounds for future pharmacokinetic and pharmacological studies. Full article

Background/Objectives: Tripterygium glycosides (TG) are used to treat inflammatory and autoimmune diseases, but their clinical application is limited by toxicity and the lack of process-responsive quality markers. This study examined whether roasting and dealkalization remodel the TG metabolite profile and alter the post-dose serum profile of circulating prototype constituents. Methods: Self-prepared TG, roasted TG (RTG), roasted–dealkalized TG (RDTG), and five marketed products were profiled by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Seven representative compounds were quantified by validated high-performance liquid chromatography (HPLC). Rat serum after oral administration was analyzed to compare circulating prototype constituents. Results: We characterized 243 constituents in material samples and 63 circulating prototype constituents in serum. Roasting primarily reshapes the profiles of diterpenoids and triterpenoids. Celastrol was not detected in the RTG and RDTG material samples, nor in the corresponding single-time-point serum profiles under the current analytical conditions. In contrast, wilforlide A exhibited an increase in material samples. Dealkalization preferentially reduced alkaloid-related constituents, including wilforine in material samples and tripterygiumine T in serum. Conclusions: Integrated material profiling, targeted quantification, and serum prototype analysis identified candidate process-responsive markers for processed TG preparations. Because the serum study was based on relative signal intensities rather than full pharmacokinetics, these markers require further pharmacokinetic and toxicological validation. Full article

Radix Paeoniae Alba, the dried root of Paeonia lactiflora Pall., contains characteristic monoterpene glycosides, but efficient recovery of these polar constituents remains challenging. This study developed an ionic liquid-assisted sequential ultrasound–microwave extraction method and evaluated paeoniflorin, oxypaeoniflorin and albiflorin by HPLC as multi-marker responses. Among the ionic liquids tested, 1-propyl-3-methylimidazolium dihydrogen phosphate showed the best extraction response. Box–Behnken response surface optimization gave practical extraction conditions of a solid-to-liquid ratio of 1:26 g/mL, ionic liquid concentration of 0.12 mol/L and ultrasound time of 22 min. Under these conditions, paeoniflorin and total marker glycosides reached 29.12 and 34.98 mg/g dry material, respectively, representing increases of 32.4% and 34.5% compared with conventional reflux extraction. UPLC-QTOF-MS profiling provided complementary chemical profile information for the optimized extract and tentatively annotated Paeonia-related monoterpene glycoside derivatives, galloylated glucose derivatives and polyphenolic constituents. Electrostatic potential, SAPT and non-covalent interaction analyses, supported by ¹H NMR chemical shift perturbation, suggested possible hydrogen bonding, electrostatic and dispersion interactions between paeoniflorin and the selected ionic liquid. These results support the optimized process as an efficient extraction approach and provide molecular interaction insights into ionic liquid-assisted recovery of monoterpene glycosides. Full article

Zanthoxylum bungeanum alkaloids have attracted considerable attention for their potential health benefits, yet systematic investigations into their extraction, purification, and comprehensive bioactivity remain scarce. In this study, an efficient extraction protocol was developed and scaled up, followed by purification using macroporous resin NKA-9 with 70% ethanol as the optimal eluent. Ultra-performance liquid chromatography coupled with ion mobility quadrupole time-of-flight mass spectrometry (UPLC-IM-QTOF/MS) identified 11 major alkaloids in the 70% ethanol eluate fraction. The antioxidant capacity of the 70% ethanol eluate fraction was evaluated through DPPH, ABTS⁺·, and Fe³⁺ reducing power assays, revealing a clear dose-dependent effect, albeit weaker than ascorbic acid. Antibacterial screening against four pathogenic bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa) via the Oxford cup method, macrobroth dilution, and minimum bactericidal concentration (MBC) determination demonstrated strain-selective activity. The strongest effect was observed against E. coli [minimum inhibitory concentration (MIC) = 2 mg/mL, MBC = 8 mg/mL, MBC/MIC = 4, bactericidal], followed by moderate activity against B. subtilis (MIC = 4 mg/mL, MBC = 16 mg/mL), while only bacteriostatic effects were noted against S. aureus and P. aeruginosa within the tested range. These findings provide a robust foundation for further development of Zanthoxylum bungeanum alkaloids as natural functional ingredients or food-compatible bio preservatives. Full article

Cholestatic liver injury (CLI) is a complex condition for which current treatment options remain limited. Lysimachiae Herba (LH), a traditional Chinese medicine with hepatoprotective properties, has not yet been fully characterized in terms of its active constituents and underlying mechanisms in CLI. This study was designed to systematically determine the chemical composition of LH, characterize its absorbed constituents in vivo, and elucidate its therapeutic mechanisms against CLI. UPLC-Q-TOF-MS/MS was employed to analyze the chemical composition of LH and its absorbed components in rat serum. Key targets and signaling pathways were predicted using network pharmacology and molecular docking, followed by experimental validation in an ANIT-induced CLI mouse model and LCA-treated HepG₂ cells through biochemical assays, histological examination, transcriptomic analysis, qRT-PCR, Western blotting, and immunofluorescence analysis. A total of 129 compounds were tentatively identified in LH, among which 26 were detected in the bloodstream. Network analysis and molecular docking suggested that LH regulates bile acid homeostasis predominantly by the FXR signaling pathway. Both in vivo and in vitro experiments provided convergent evidence that LH modulates the FXR-related bile acid regulatory network, enhances bile acid efflux transporter expression, and alleviates CLI. In conclusion, this study systematically elucidates the chemical composition, absorbed constituents, and pharmacological mechanisms of LH in CLI, highlighting the involvement of FXR-related bile acid regulation as an important mechanism and providing a scientific basis for the potential development of LH for cholestatic liver injury. Full article