Search Results (48)

Background/Objectives: Hyperuricemia is a metabolic disorder characterized by renal dysfunction and systemic inflammation. While Clematis chinensis Osbeck is traditionally used for gout-related conditions, its chemical basis and precise mechanisms remain poorly understood. This study aimed to characterize the bioactive fraction (CWE-60EF) and elucidate its multi-target regulatory mechanisms against hyperuricemia. Methods: Qualitative and quantitative chemical profiling of CWE-60EF was performed using high-resolution LC-MS/MS. Its anti-hyperuricemic activity was validated using in vitro xanthine oxidase (XOD) inhibition, a zebrafish model, and HK-2 cell injury models. Mechanisms were explored through an integrated approach combining bioinformatics, Mendelian randomization (MR), and molecular docking. Results: A total of 50 compounds, primarily alkaloids and flavonoids (e.g., magnoflorine and phloretin), were characterized in CWE-60EF, and major marker compounds were quantitatively standardized. The fraction significantly inhibited XOD activity and rescued hyperuricemia-associated phenotypes in zebrafish. In HK-2 cells, CWE-60EF suppressed adenosine- and urate-induced cellular injury and the transcriptional expression of pro-inflammatory cytokines (IL-6 and IL-1β). MR analysis provided genetic evidence supporting IL-6 as a causal mediator of gout risk. Integrative analysis revealed that the protective effects of CWE-60EF are mediated through the coordinated regulation of purine metabolism, inflammatory cascades, and urate transporters (URAT1/GLUT9). Conclusions: This study demonstrates that CWE-60EF is a quantitatively standardized bioactive fraction that exerts anti-hyperuricemic, renoprotective, and anti-inflammatory effects by modulating uric acid metabolism and inflammation. By integrating genetic causality with phytochemical validation, our findings provide a novel mechanistic foundation for the traditional application of C. chinensis in hyperuricemic disorders. Full article

The objective of this paper was to examine the effects of differing levels of dietary Zanthoxylum bungeanum seed cake and meal (ZBS) on the productive performance, egg quality, gut microbiota and liver metabolomics of laying hens. In total, 1280 healthy Lohmann Pink laying hens (age, 36 weeks) were randomly assigned to four treatment groups: control (CON), 1% ZBS, 2% ZBS, or 3% ZBS. The trial lasted for eight weeks. The results indicated that compared to the CON group, the 3% ZBS group had a higher egg production during weeks 5–8 and 1–8 (p < 0.05). However, the feed-to-egg ratio and egg breakage rate during weeks 1–8 was lower in the 2% ZBS group and 3% ZBS group compared to the CON group (p < 0.05). Adding different levels of ZBS to the diet significantly increased egg production, reduced the feed-to-egg ratio, and reduced the feed intake of weeks 5–8 in laying hens (p < 0.05) with both effects exhibiting a linear relationship as the addition level increased (p < 0.05). Compared to the CON group, supplementing the diet with 2% or 3% ZBS significantly increased T-AOC levels in the liver and serum, as well as T-SOD enzyme activity, but it significantly decreased the MDA level in the serum (p < 0.05). The addition of ZBS has been demonstrated to induce alterations in the microbial composition of the cecum in laying hens. The key metabolic pathways and key metabolites are glycerophospholipid metabolism and magnoflorine, respectively. In conclusion, the supplementation of diets with ZBS has been demonstrated to significantly enhance the production performance and antioxidant capacity in laying hens. The optimal supplementation level was 3% ZBS. Full article

Astrocytes play a crucial role in maintaining neuronal microenvironment homeostasis and regulating synaptic plasticity within the hippocampus. Magnoflorine (MGN), a naturally occurring isoquinoline alkaloid, has demonstrated biological activity in the central nervous system. However, its effects on astroglial cells remain poorly understood. The present study aimed to evaluate the impact of acute and chronic administration of MGN (10 and 20 mg/kg body weight) on the morphology and morphometric parameters of GFAP-positive astrocytes in the CA1 field of the mouse hippocampus. Immunohistochemical and morphometric analyses were performed in the oriens layer (SO), pyramidal layer (SP), radiate layer (SR), and lacunose-molecular layer (SLM). MGN significantly modulated astrocyte density, cell size, and the number of processes in a dose-, time-, and layer-dependent manner. A heterogeneous and layer-specific astroglial response was particularly evident following chronic administration of the tested compound. Together with the observed lack of significant differences in analysed parameters, decreases were mainly detected after administration of the low MGN dose, whereas the 20 mg/kg dose induced primarily increased structural complexity. Thus, the direction of changes was not uniform across all layers. The most prominent changes were detected in the SLM layer. Overall, MGN modulated astrocyte morphology and reactivity in a context-dependent manner. These findings indicate a modulatory influence of MGN on astroglial structural plasticity rather than a uniform directional effect. Although the observed changes may be associated with alterations in astroglia-mediated mechanisms involved in maintaining neuronal homeostasis and responses to stress, their functional significance requires further investigation. Full article

The Cinnamomum camphora seed kernel (CCSK) shows great promise as a natural source of bioactive alkaloids. However, there is little data about recovering alkaloids from CCSK by-products after oil extraction using an aqueous method. This study aimed to establish an efficient technology for enriching CCSK alkaloids (including magnoflorine, lindoldhamine and N,N-methyldomesticinium) using macroporous resin technology. The results showed that XR918C resin was the most suitable adsorbent due to its high adsorption/desorption capacity for CCSK alkaloids. The adsorption process was best described by Langmuir isotherm models and pseudo-second-order kinetics; it was spontaneous and physical in nature. The optimum procedure for CCSK alkaloids enrichment using XR918C resin was as follows: for adsorption, the injection flow rate and sample volume were 2.0 BV/h and 7.0 BV, respectively; for desorption, the eluent type, elution flow rate and volume were 80% ethanol, 2.0 BV/h and 6.0 BV, respectively. Furthermore, the scale-up of the CCSK alkaloid enrichment process was performed under optimal conditions. Following the 10-fold scale-up enrichment, the content of CCSK alkaloids was raised 4.41-fold, with a recovery rate of 89.19 ± 0.01%. After nine regeneration cycles, the efficiency of the XR918C resin remained stable, indicating its good reusability. In addition, CCSK alkaloids exhibited strong in vitro antioxidant activity. This study provides a useful reference for the industrial-scale enrichment of CCSK alkaloids. Full article

Hypecoum erectum L. is a medicinal plant known for its high content of isoquinoline alkaloids (IQAs), a class of compounds with diverse pharmacological activities. To elucidate the biosynthetic mechanisms and tissue-specific accumulation of IQAs, we integrated HPLC-MS/MS-based metabolomic analysis with RNA sequencing (RNA-seq) transcriptomic profiling across the roots, stems, and leaves of H. erectum. Metabolomic analysis identified twenty-six IQAs as differentially accumulated metabolites (DAMs) among the three tissues, while transcriptomic analysis revealed twenty-two categories of differentially expressed genes (DEGs) involved in IQA biosynthesis. KEGG pathway enrichment analysis demonstrated that nine DAMs and twenty categories of DEGs were co-enriched in the IQA biosynthetic pathway of Hypecoum erectum. Notably, seven key DAMs—Stylopine, Protopine, Magnoflorine, Corydaline, Tetrahydropalmatine, Sanguinarine, and Palmatine—preferentially accumulated in the root, concomitant with the elevated expression of eleven root-specific DEGs, including GOT1, CYP719A14, SMT, CYP719A1_2_3_13, PSOMT1, E2.1.1.116, CYP80B1, E2.1.1.128, NCS, ASP5, and BBE1. Gene–metabolite correlation network analysis further identified nine DAMs and fifteen DEGs closely associated with IQA biosynthesis, highlighting key enzymatic regulators of alkaloid accumulation. Additionally, several transcription factor (TF) families, including bHLH, NAC, and ERF families, were predicted to participate in the transcriptional regulation of IQA-related genes. Collectively, these findings demonstrate that roots are the primary site of IQA biosynthesis in H. erectum and provide a molecular framework for understanding the regulation and utilization of its medicinally active components. Full article

Ziziphi Spinosae Semen (ZSS), a medicinal and edible homologous herbal drug, is commercially available in both raw and fried (FZSS) forms and has been widely used to improve sleep quality. This study aimed to elucidate the differences in chemical composition between the two specifications. A comprehensive metabolomics approach utilizing ¹H NMR, GC–MS, and UHPLC-Q-Orbitrap-HRMS identified a total of 66 potential biomarkers. The results demonstrated that after frying, the content of fatty acids decreased significantly, while the levels of most primary metabolites (e.g., sugars, amino acids) and secondary metabolites (e.g., alkaloids, flavonoids) increased markedly. Targeted quantification of 14 key components validated these trends: the contents of five fatty acids decreased (p < 0.001), whereas the levels of five secondary metabolites (coclaurine, magnoflorine, spinosin, 6‴-feruloylspinosin, and jujuboside A) increased. In contrast, the content of jujuboside B decreased significantly. This study systematically reveals the profound impact of frying on the chemical composition of ZSS, providing a scientific basis for its quality control and processing optimization. Full article

Background/Objectives: Bangkeehwangkee-tang (BHT) is a traditional herbal formula composed of six medicinal herbs: Sinomenii Caulis et Rhizoma, Astragali Radix, Atractylodis Rhizoma Alba, Zingiberis Rhizoma Recens, Zizyphi Fructus, and Glycyrrhizae Radix et Rhizoma. BHT has been widely used for its immunomodulatory and anti-inflammatory effects. This study aimed to develop a reliable analytical method for the simultaneous determination of 22 marker compounds to ensure consistent quality control and to ensure consistent efficacy in both clinical and non-clinical studies of BHT. Methods: An ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method based on multiple reaction monitoring was developed and validated for the simultaneous determination of 22 marker compounds in BHT. The method was evaluated for selectivity, linearity (coefficient of determination, r²), sensitivity (limit of detection (LOD) and limit of quantification (LOQ)), accuracy (recovery), and precision (relative standard deviation (RSD)) in accordance with guidelines. Results: The developed method exhibited excellent selectivity and linearity (r² ≥ 0.9913) for all target compounds. The LOD and LOQ ranged from 0.09 μg/L to 326.58 μg/L and 0.28 μg/L to 979.75 μg/L, respectively. The recovery ranged from 90.36% to 113.74%, and precision (RSD) was ≤15%, confirming the method’s reliability. The application of the method to various BHT samples revealed substantial variations in the marker compound contents, particularly for sinomenine, magnoflorine, and glycyrrhizin. Conclusions: These findings highlight the necessity for standardized quality control of BHT and demonstrate that the developed UPLC–MS/MS method is a practical and reliable tool for performing quality assessment of traditional herbal formulas. Full article

Background: Rheumatoid arthritis (RA) is a chronic inflammatory disorder characterized by synovial hyperplasia and joint destruction. Previous studies have demonstrated that the alkaloids of Rushanhu (ARSHs), the dried root and stem of Zanthoxylum nitidum var. tomentosum, exhibit favorable therapeutic effects on RA, and this study aims to investigate the underlying molecular mechanisms involved. Methods: A complete Freund’s adjuvant (CFA)-induced arthritis model in male SD rats (n = 64) was used to evaluate ARSHs. Groups included control, model, methotrexate (MTX), and ARSH-treated. Therapeutic effects were assessed via arthritis index, paw swelling, and serum cytokines (IL-1β, IL-6, IL-17A). Network pharmacology identified bioactive alkaloids and core targets, validated by molecular docking. In vitro mechanisms (proliferation, apoptosis, signaling pathways) were examined in MH7A synovial cells. Results: ARSHs significantly attenuated joint inflammation and damage in CFA rats (* p < 0.01 vs. model), reducing pro-inflammatory cytokines. Fifteen alkaloids (e.g., dihydrochelerythrine, magnoflorine) and 24 targets (e.g., SRC, STAT3, MAPK3) were prioritized. Molecular docking confirmed strong binding (binding energy < −7.0 kcal/mol). In vitro, ARSHs suppressed MH7A proliferation and induced apoptosis via Bcl-2/Bax dysregulation and the inhibition of SRC/STAT3/MAPK3 phosphorylation. Conclusions: ARSHs mitigate RA pathogenesis by targeting the SRC/STAT3/MAPK3 signaling axis in synovial cells. This study provides mechanistic validation of ARSHs as multi-target phytotherapeutic agents against inflammatory arthritis. Full article

Chinese herbal medicines have played a significant role in the development of new and effective drugs, but how to identify the active ingredients from complex extracts of traditional Chinese herbal medicines was a research difficulty. In recent years, few studies have focused on high-efficiency identification of small-molecule inhibitors of Programmed Death Ligand 1 with lower antigenicity and flexible structure tunability. In order to identify small molecule inhibitors of PD-L1 from complex Chinese herbal extracts, this study established a protein–ligand trapping system based on high-performance liquid chromatography coupled with a photo-diode array detector, ion trap/quadrupole time-of-flight tandem mass spectrometry, and a Programmed Death Ligand 1 affinity chromatography unit (ACPD-L1-HPLC-PDA-IT-TOF (Q-TOF)-MS) to rapidly screen and identify small-molecule inhibitors of Programmed Death Ligand 1 from Toddalia asiatica (L.) Lam. Fourteen components were then identified as PD-L1 binders, and surface plasmon resonance (SPR) validation results showed that six of them—magnoflorine (6), nitidine (22), chelerythrine (24), jatrorrhizine (13), toddaculin (68), and toddanol (45)—displayed PD-L1 binding activity. Laser scanning confocal microscopy results demonstrated that these compounds effectively inhibited the binding of PD-1 to PD-L1 in a dose-dependent manner. Additionally, flow cytometry analysis indicated they could promote human lung cancer cell line (A549) apoptosis when co-cultured with Peripheral Blood Mononuclear Cells (PBMCs). The system’s innovation lies in its first integration of dynamic protein–ligand trapping with multi-dimensional validation, coupled with high-throughput screening capacity for structurally diverse natural products. This workflow overcomes traditional phytochemical screening bottlenecks by preserving native protein conformations during affinity capture while maintaining chromatographic resolution, offering a transformative template for accelerating natural product-derived immunotherapeutics through the PD-1/PD-L1 pathway. Full article

Background: The postmortem diagnosis of fatal hypothermia presents a considerable challenge in forensic medicine. Metabolomics, a powerful tool reflecting comprehensive changes in endogenous metabolites, offers significant potential for exploring disease mechanisms and identifying diagnostic markers. Methods: In this study, we employed ultra-high-performance liquid chromatography–mass spectrometry (UHPLC–MS) to perform a non-targeted metabolomic analysis of liver, stomach, spleen, and musculus gastrocnemius tissues from mice subjected to fatal hypothermia. Result: A substantial number of differential metabolites were identified in each tissue: 1601 in the liver, 420 in the stomach, 732 in the spleen, and 668 in the gastrocnemius muscle. The most significantly altered metabolites were as follows: magnoflorine (liver, upregulated, ranked first in fold-change), gibberellic acid (stomach, downregulated, ranked first in fold-change), nitrofurantoin (spleen, upregulated, ranked first in fold-change), and isoreserpin (gastrocnemius muscle, downregulated, ranked first in fold-change). Glycerophospholipid metabolism exhibited notable enrichment in all tissues (spleen: second, liver: tenth, stomach: eleventh, gastrocnemius muscle: twenty-first), as did tryptophan metabolism (spleen: thirteenth, liver: eighth, stomach: third, gastrocnemius muscle: seventeenth). Conclusions: Our findings provide insights into the metabolic perturbations associated with fatal hypothermia in different tissues and lay a foundation for the identification of potential tissue biomarkers for forensic diagnosis. Full article

Background/Objectives: We assessed the influence of long-term injection of magnoflorine (MAG) on memory acquisition in mice for the first time. Methods: This isoquinoline alkaloid that belongs to the aporphines was isolated from the roots of Berberis vulgaris by centrifugal partition chromatography (CPC) using a biphasic solvent system composed of chloroform: methanol: water in the ratio 4:3:3 (v/v/v) with 20 mM of hydrochloric acid and triethylamine, within 64 min. Results: Our results indicated that long-term injection of MAG 20 mg/kg dose improve the long-term memory acquisition in mice that were evaluated in the passive avoidance (PA) test with no toxicity records. The analysis of brain lysates and animal plasma by HPLC-ESI-QTOF-MS/MS showed the ability of the compound to cross the blood–brain barrier, and an elevated level of phosphatidylcholine PC (14:1(9Z)/14:1(9Z)) with the molecular formula of C₃₆H₆₉NO₈P was observed in both treated groups with 10 mg/kg and 20 mg/kg MAG in comparison to the control group. Conclusions: This phenomenon may explain MAG’s cognition-enhancing properties as the PC may induce the synthesis and strengthening of neuronal cells. Also, the 7-day-long administration of MAG at 10 mg/kg and 20 mg/kg increased the mean number of parvalbumin (PV)-IR neurons in the hippocampus. Statistically, the largest PV-IR neurons were observed at the 20 mg/kg dose, which may indicate a potential effect of MAG on Ca²⁺ metabolism. However, no statistical differences were observed in the mean number of PV-IR nerve fibers in both doses of MAG, regardless of the hippocampal fields. This positive effect of MAG on hippocampal neurons provides further support for the neuroprotective effect of this alkaloid. Full article

Lung cancer is a prevalent and very aggressive sickness that will likely claim 1.8 million lives by 2022, with an estimated 2.2 million additional cases expected worldwide. The goal of the current investigation was to determine whether petroleum ether extract of purslane leaf could be used to treat lung cancer induced by 4-(Methylnitrosamino)-1-(3-pyridyl)-1-buta-4 none (NNK) in rats. In the in vitro extract recorded, promising anticancer effects in A540 cell lines with IC₅₀ were close to the reference drug, doxorubicin (14.3 and 13.8 μg/mL, respectively). A dose of 500 mg/kg/day orally for 20 weeks exhibited recovery effects on NNK-induced lung cancer with a good safety margin, where Intercellular Adhesion Molecule-1 (ICAM-1), the lung cancer biomarker, was significantly reduced by about 18.75% compared to cancer control. Purslane exhibited many anticancer mechanisms, including (i) anti-proliferation as a significant reduction in Ki67 level (20.42%), (ii) anti-angiogenesis as evident by a considerable decrease in Matrix metalloproteinase-9 (MMP-9) expression (79%), (iii) anti-inflammation as a remarked decline in Insulin-like growth factor 1 (IGF-1) expression (62%), (iv) pro-apoptotic effect as a significant activation in Forkhead box protein O1 (FOXO1) expression (262%), and (v) anti-oxidation as remarkable activation on antioxidant biomarkers either non-enzymatic or enzymatic concurrent with considerable depletion on oxidative stress biomarker, in comparison to cancer control. The histopathological examination revealed that Purslane extract showed markedly improved tissue structure and reduced pathological changes across all examined organs caused by NNK. The anti-lung cancer effect exhibited by the extract may be linked to the active ingredients of the extract that were characterized by LC–MS, such as α-linolenic acid, linoleic acid, palmitic acid, β-sitosterol, and alkaloids (berberine and magnoflorine). Full article

A low-cost method for the simultaneous extraction of alkaloids and water-insoluble flavonoids and esters from Coptis chinensis Franch. (Abbrev. C. chinensis) was explored to provide a reference for the production of green plant-based preparations and traditional Chinese medicine formula granules. A combined extraction method with the deep eutectic solvents (DESs) of choline chloride and urea (molar mass ratio of 1:2) and organic solvent ethanol was used, supplemented by ultrasonic-assisted extraction (ultrasonic power: 150 W; ultrasonic temperature: 60 °C; treatment time: 15 min). The extraction efficiency of the 50% DES (choline chloride and urea) aqueous solution for berberine, palmatine, jatrorrhizine, and magnoflorine was found to be the highest and was superior to traditional ultrasonic extraction and water bath reflux extraction methods. Furthermore, the flavonoids and esters from C. chinensis residue were extracted using ethanol. The results from high-performance liquid chromatography and gas chromatography–mass spectrometry indicated a high extraction efficiency overall. Full article

Cold stress affects the seed germination and early growth of winter rapeseed, leading to yield losses. We employed transmission electron microscopy, physiological analyses, metabolome profiling, and transcriptome sequencing to understand the effect of cold stress (0 °C, LW) on the cotyledons of cold-tolerant (GX74) and -sensitive (XY15) rapeseeds. The mesophyll cells in cold-treated XY15 were severely damaged compared to slightly damaged cells in GX74. The fructose, glucose, malondialdehyde, and proline contents increased after cold stress in both genotypes; however, GX74 had significantly higher content than XY15. The pyruvic acid content increased after cold stress in GX74, but decreased in XY15. Metabolome analysis detected 590 compounds, of which 32 and 74 were differentially accumulated in GX74 (CK vs. cold stress) and XY15 (CK vs. cold stressed). Arachidonic acid and magnoflorine were the most up-accumulated metabolites in GX74 subjected to cold stress compared to CK. There were 461 and 1481 differentially expressed genes (DEGs) specific to XY15 and GX74 rapeseeds, respectively. Generally, the commonly expressed genes had higher expressions in GX74 compared to XY15 in CK and cold stress conditions. The expression changes in DEGs related to photosynthesis-antenna proteins, chlorophyll biosynthesis, and sugar biosynthesis-related pathways were consistent with the fructose and glucose levels in cotyledons. Compared to XY15, GX74 showed upregulation of a higher number of genes/transcripts related to arachidonic acid, pyruvic acid, arginine and proline biosynthesis, cell wall changes, reactive oxygen species scavenging, cold-responsive pathways, and phytohormone-related pathways. Taken together, our results provide a detailed overview of the cold stress responses in rapeseed cotyledons. Full article

A robust, well-functioning immune system is the cornerstone of good health. Various factors may influence the immune system’s effectiveness, potentially leading to immune system failure. This review aims to provide an overview of the structure and action of immunomodulators isolated from African medicinal plants. The research was conducted according to PRISMA guidelines. Full-text access research articles published in English up to December 2023, including plant characteristics, isolated phytochemicals, and immuno-modulatory activities, were screened. The chemical structures of the isolated compounds were generated using ChemDraw^® (version 12.0.1076), and convergent and distinctive signaling pathways were highlighted. These phytochemicals with demonstrated immunostimulatory activity include alkaloids (berberine, piperine, magnoflorine), polysaccharides (pectin, glucan, acemannan, CALB-4, GMP90-1), glycosides (syringin, cordifolioside, tinocordiside, aucubin), phenolic compounds (ferulic acid, vanillic acid, eupalitin), flavonoids (curcumin, centaurein, kaempferin, luteolin, guajaverin, etc.), terpenoids (oleanolic acid, ursolic acid, betulinic acid, boswellic acids, corosolic acid, nimbidin, andrographolides). These discussed compounds exert their effects through various mechanisms, targeting the modulation of MAPKs, PI3K-Akt, and NF-kB. These mechanisms can support the traditional use of medicinal plants to treat immune-related diseases. The outcomes of this overview are to provoke structural action optimization, to orient research on particular natural chemicals for managing inflammatory, infectious diseases and cancers, or to boost vaccine immunogenicity. Full article