ETHNOPHARMACOLOGICAL RELEVANCE: Rhododendron dauricum L. is used in traditional medicine for respiratory disorders, but its developmental and cardiac safety remains insufficiently defined. AIM OF THE STUDY: To characteri...ETHNOPHARMACOLOGICAL RELEVANCE: Rhododendron dauricum L. is used in traditional medicine for respiratory disorders, but its developmental and cardiac safety remains insufficiently defined. AIM OF THE STUDY: To characterize developmental/cardiac phenotypes induced by Rhododendron dauricum hot-water extract (RDWE), identify toxicity-enriched fractions and candidate constituents, and assess whether selective fraction removal reduces toxicity-related effects. MATERIALS AND METHODS: Zebrafish embryos/larvae were exposed to RDWE, four membrane fractions, and candidate monomers. LC50 values were calculated from 96 hpf mortality data using three replicate wells per concentration and 10 embryos per well. Candidate monomers were tested over 0-200 mg/L; rhododendrol-associated responses were examined at 0-40 mg/L using RNA-seq, Fluo-4, ROS, AO staining, and qRT-PCR. RESULTS: RDWE induced concentration-dependent developmental/cardiac abnormalities, mainly pericardial and yolk-sac edema, with a 96 hpf LC50 of 228.5 mg/L (95% CI, 181.8-276.2). The 5-10 kDa fraction showed the highest mortality-based potency among fractions (80.68 mg/L; 95% CI, 66.70-94.97). Rhododendrol showed the lowest LC50 among candidate monomers (67.82 mg/L; 95% CI, 55.65-81.49) and induced pericardial edema and bradycardia. Rhododendrol-associated cardiotoxicity was accompanied by calcium accumulation, oxidative stress, apoptosis-related signals, calcium-signaling enrichment, down-regulation of cacna1ab, cacna1da, atp2a1l, and cat, and up-regulation of cu/zn-sod and caspase3/8/9. Removing the 5-10 kDa fraction increased the LC50 to 321.0 mg/L (95% CI, 299.0-347.9). CONCLUSIONS: The 5-10 kDa fraction was identified as a toxicity-enriched fraction, and rhododendrol was identified as a potential key contributor to toxicity. Selective removal of this fraction may represent a potential toxicity-reduction strategy, but further validation is required to determine whether pharmacological activity, chemical consistency, and mammalian relevance are retained after fraction removal.
ETHNOPHARMACOLOGICAL RELEVANCE: The nine steaming and nine sun-drying method, a classic representative of traditional Chinese medicine (TCM) processing, is primarily used for processing precious tonic herbs. Its core val...ETHNOPHARMACOLOGICAL RELEVANCE: The nine steaming and nine sun-drying method, a classic representative of traditional Chinese medicine (TCM) processing, is primarily used for processing precious tonic herbs. Its core value lies in its multi-faceted regulation of herbal properties, thereby providing a material basis for TCM syndrome differentiation and treatment. AIM OF THE REVIEW: This review aims to systematically summarize the historical evolution and technical characteristics of this processing method, integrate representative herbs under a unified analytical framework, and interpret this traditional processing method from the perspectives of coordinated chemical transformation, bioactivity remodeling, and modernization potential. MATERIALS AND METHODS: Available literature related to the nine steaming and nine sun-drying process was collected by searching PubMed, ScienceDirect, Web of Science, Scopus, and CNKI. RESULTS: Through repeated steaming and sun-drying, this process markedly alters the chemical composition of selected medicinal materials, such as Heshouwu, Huangjing, rhubarb, and Rehmanniae Radix. These transformations, mediated by mechanisms including pyrolysis and hydrolysis, oxidation-reduction, and Maillard reactions, may optimize the component profile and contribute to enhanced pharmacological activities (e.g., immunomodulation, neuroprotection, and hematinic effects), while attenuating toxicity-related or irritant effects in specific herbs, such as hepatotoxicity-related constituents in Heshouwu and gastrointestinal irritation in Huangjing or rhubarb. CONCLUSIONS: The nine steaming and nine sun-drying process achieves the optimization of herbal properties and potentiation of therapeutic effects, but challenges such as low production efficiency and difficulties in quality control persist in its modern application. In-depth research is expected to promote its inheritance and innovation.
ETHNOPHARMACOLOGICAL RELEVANCE: Chemotherapy-induced myelosuppression is a common and severe complication in clinical oncology treatment. Chemotherapeutic agents trigger this condition by damaging bone marrow mesenchymal...ETHNOPHARMACOLOGICAL RELEVANCE: Chemotherapy-induced myelosuppression is a common and severe complication in clinical oncology treatment. Chemotherapeutic agents trigger this condition by damaging bone marrow mesenchymal stem cells (BMSCs) and disrupting hematopoietic microenvironment homeostasis. Danggui Jixueteng Decoction (DJD), a classical blood-tonifying and blood-activating formula, has been confirmed to alleviate post-chemotherapy myelosuppression, but its specific molecular mechanisms in protecting BMSCs remain unclear. OBJECTIVE: This study aims to clarify the molecular mechanisms whereby DJD-containing serum alleviates chemotherapy-induced BMSCs injury, identify key signaling pathways and targets for its myeloprotection, and provide evidence supporting DJD for treating chemotherapy-induced myelosuppression. MATERIALS AND METHODS: UHPLC-Orbitrap-MS analyzed DJD's chemical constituents, including blood-entry prototypes and metabolites. A carboplatin (CBP) -induced cell injury model was established. Different concentrations of DJD-containing serum were applied. Functional, proteomic, Western blot, molecular docking, cellular thermal shift assay (CETSA) and molecular dynamics simulation (MD) were used for detection and verification. RESULTS: 107 chemical components and 38 blood-entry components of DJD were identified. DJD-containing serum concentration-dependently reversed the aforementioned damaging effects of CBP, significantly improved mitochondrial function, suppressed excessive mitophagy and ferroptosis, and effectively restored AKT/FOXO3a pathway signaling function. Molecular docking, CETSA and MD validated that tanshinol B stably binds to FOXO3a and enhances its thermal stability. CONCLUSION: This study demonstrates that DJD-containing serum protects BMSCs and maintains hematopoietic microenvironment homeostasis by targeting the AKT/FOXO3a signaling pathway, thereby repairing its abnormally inhibited state, suppressing CBP-induced excessive mitophagy, and inhibiting subsequent ferroptosis. This study provides novel insights and experimental evidence for targeted intervention of chemotherapy-induced bone marrow injury using traditional Chinese medicine.
Panti A, Fusi F, De Rosa F
… +11 more, Sommella EM, Carullo G, Strangis EF, Aiello F, Aloi A, Gentile M, Campiani G, Puoci F, Logozzi M, Valoti M, Saponara S
ETHNOPHARMACOLOGICAL RELEVANCE: Grapevine leaves are traditionally used in Mediterranean and Middle Eastern ethnomedicine to treat circulatory disorders, inflammation, and venous insufficiency. However, cultivar-specific...ETHNOPHARMACOLOGICAL RELEVANCE: Grapevine leaves are traditionally used in Mediterranean and Middle Eastern ethnomedicine to treat circulatory disorders, inflammation, and venous insufficiency. However, cultivar-specific phytochemical profiles and their underlying vascular mechanisms remain poorly characterised. AIM OF THE STUDY: To investigate the chemical composition and vascular activity of extracts and extracellular micro- and nanovesicles (EVs) derived from leaves of the Italian Vitis vinifera cv. Sagrantino. MATERIALS AND METHODS: Extracts were obtained using ultrasound-assisted extraction and Soxhlet methods with solvents of different polarity. Phytochemical profiling was performed by UHPLC-HRMS and H NMR. EVs were characterised by nanoparticle tracking analysis and transmission electron microscopy and analysed by NMR. Vascular effects were assessed ex vivo on rat aorta rings. Modulation of vascular smooth muscle Ca1.2 channels was evaluated by whole-cell patch-clamp recordings. RESULTS: Extracts were rich in polyphenols, including flavonoid glycosides, gallotannins, ellagitannins, and cinnamic acid derivatives. All extracts induced concentration-dependent vasorelaxation in endothelium-intact aorta rings, whereas removal of the endothelium markedly reduced or reversed this effect, often leading to contraction. A hormetic response was observed at higher concentrations. Ultrasound-assisted hydroalcoholic extracts showed the strongest activity. EVs showed nanoscale morphology, contained polyphenols, sugars, fatty acids and amino acids, induced vasorelaxation, and inhibited Ca1.2 channel currents in a concentration-dependent manner. CONCLUSIONS: Sagrantino grapevine leaves represent an underutilised source of vasoactive compounds. Their vascular effects involve both endothelium-dependent mechanisms and direct inhibition of Ca1.2 channels. These findings provide mechanistic support for their traditional use and highlight their potential for sustainable cardiovascular applications within a circular bioeconomy framework.
ETHNOPHARMACOLOGICAL RELEVANCE: Jinhua Qinggan Granules (JHQG) is a traditional Chinese medicine formulation widely used for the treatment of viral respiratory infections. Traditional herbal formulations contain multiple...ETHNOPHARMACOLOGICAL RELEVANCE: Jinhua Qinggan Granules (JHQG) is a traditional Chinese medicine formulation widely used for the treatment of viral respiratory infections. Traditional herbal formulations contain multiple phytochemical constituents that may act individually or synergistically to produce therapeutic effects. Although its clinical applications have been reported, the bioactive constituents underlying its antiviral activity and their mechanisms of action remain to be clarified. AIM OF THE STUDY: This study aimed to identify representative bioactive constituents of JHQG with antiviral activity against Zika virus (ZIKV) and to investigate their potential mechanisms of action. MATERIALS AND METHODS: A panel of representative phytochemical constituents reported in JHQG was screened for antiviral activity against ZIKV in vitro. Antiviral efficacy and cytotoxicity were evaluated by determining EC and CC values. Viral replication was assessed using plaque assay, RT-qPCR, immunofluorescence, and Western blot analysis. Time-of-addition experiments were performed to explore stage-specific effects. Transcriptome analysis and quantitative PCR were conducted to examine host inflammatory gene expression. Intracellular reactive oxygen species (ROS) levels and NF-κB activation were evaluated in stimulated cells. In vivo antiviral efficacy was assessed in a ZIKV infection model using A129 mice. Molecular docking and molecular dynamics simulations were performed to explore potential interactions with the viral NS3 protease. RESULTS: Among the screened compounds, chlorogenic acid (CGA) exhibited notable antiviral activity and was selected for further investigation. CGA treatment was associated with reduced ZIKV replication, decreased viral protein expression, and lower production of infectious viral particles under the tested conditions. Time-of-addition experiments indicated that CGA exerted inhibitory effects during early stages and throughout the infection period. ZIKV infection induced the expression of inflammatory genes including CXCL8, CXCL10, and TNFAIP3, whereas treatment with JHQG or CGA was associated with reduced expression of these genes. CGA treatment also corresponded with decreased intracellular ROS levels and attenuation of NF-κB activation. In A129 mice, CGA administration was associated with improved survival, reduced viral RNA levels in brain and liver tissues, and partial preservation of tissue morphology. Computational analyses suggested potential interactions between CGA and the NS3 protease. CONCLUSION: These findings identify CGA as a representative bioactive constituent selected from the screened phytochemical components of JHQG that may contribute to its antiviral activity. The antiviral effects observed in this study, together with the modulation of host inflammatory responses, provide experimental evidence supporting further investigation of CGA as a candidate compound for the development of therapeutic strategies against ZIKV infection.
UNLABELLED: Polycystic ovary syndrome (PCOS) is a multifactorial endocrine disorder characterized by ovulation disorders. Zishen Qingre Lishi Huayu Recipe (ZQLHR), a traditional Chinese medicine (TCM) formula, has shown...UNLABELLED: Polycystic ovary syndrome (PCOS) is a multifactorial endocrine disorder characterized by ovulation disorders. Zishen Qingre Lishi Huayu Recipe (ZQLHR), a traditional Chinese medicine (TCM) formula, has shown clinical efficacy by effectively increasing the ovulation rate and menstrual cycle regularity in patients with PCOS and is widely used in practice; however, its underlying pharmacological mechanism has not been fully elucidated. AIM OF THE STUDY: Aim of the study: This study aimed to elucidate how ZQLHR modulates the proliferation-apoptosis balance of ovarian granulosa cells (GCs) in PCOS via the C/EBPβ/STAT3 signaling axis. METHODS: Human granulosa cells (hGCs) from PCOS and non-PCOS patients were analyzed for proliferation, apoptosis, and C/EBPβ/STAT3 expression. In vivo, letrozole-induced PCOS mice were subjected to scAAV-mediated C/EBPβ knockdown, followed by administration of ZQLHR to evaluate treatment outcomes. In vitro, C/EBPβ or STAT3 was silenced in Human granulosa-like tumor cell line (KGN) cells to explore their regulatory relationship and roles in proliferation and apoptosis. Additionally, ZQLHR effects were evaluated in a dose- and time-dependent manner. Notably, CETSA was employed to confirm for the first time direct binding of ZQLHR to C/EBPβ, and C/EBPβ-silenced cells were subsequently treated with ZQLHR to examine whether its effects involve C/EBPβ-dependent STAT3. The pharmacological mechanism of ZQLHR was further evaluated through histological and molecular biology experiments. RESULTS: PCOS patient-derived GCs exhibited reduced proliferation, elevated apoptosis, and decreased C/EBPβ/STAT3 expression. In LET-induced PCOS mice, ZQLHR treatment significantly improved estrous cycle regularity, hormonal balance, follicular development, and GC apoptosis, accompanied by modulation of the C/EBPβ/STAT3 pathway. C/EBPβ knockdown exacerbated ovarian dysfunction and KGN apoptosis, whereas ZQLHR attenuated these effects and partially restored pathway activity. In vitro, C/EBPβ and STAT3 influenced each other and CFTR expression, associated with changes in proliferation and apoptosis. ZQLHR increased proliferation markers (PCNA, cyclin D1) and decreased apoptosis markers (Caspase-3, PARP) in a dose- and time-dependent manner, with CETSA confirming direct binding to C/EBPβ. Notably, despite C/EBPβ knockdown, ZQLHR partially restored KGN cell proliferation and modestly modulated the C/EBPβ/STAT3 signaling pathway. CONCLUSION: ZQLHR improves GC homeostasis and promotes follicular development in PCOS by modulating the C/EBPβ/STAT3 pathway.
ETHNOPHARMACOLOGICAL RELEVANCE: Rhizoma Paridis refers to the dried rhizomes of perennial herbaceous plants belonging to the genus Paris. Classified under the Melanthiaceae family, this medicinal herb has long been used...ETHNOPHARMACOLOGICAL RELEVANCE: Rhizoma Paridis refers to the dried rhizomes of perennial herbaceous plants belonging to the genus Paris. Classified under the Melanthiaceae family, this medicinal herb has long been used in folk medicine in East Asia and Europe for the treatment of bleeding, diarrhea, gastritis, mumps, abscesses, as well as snake and insect bites. AIM OF THE STUDY: This study aims to explore the anti-metastatic effects of the active fractions and phytochemicals of Paris polyphylla Smith var. yunnanensis (Franch.) Hand.-Mazz. (PPY) and Paris fargesii Franch. (PF) in vitro and in vivo, and uncover their underlying mechanisms. MATERIAL AND METHODS: MTT assays detected the cytotoxicity of extracts, active fractions, and active ingredients. To assess the suppressive effect on EC cells, Transwell and Scratch assays were employed. While Western blot assays and xenograft-bearing mouse models were utilized to explore the mechanism and the anti-metastasis effects in vitro and vivo. RESULTS: Both extracts of PPY and PF, their active fractions, and ten principal ingredients all exhibited significant cytotoxicity against EC109 and KYSE520 cells. Furthermore, the two active fractions and their shared unique steroidal glycoside 8 inhibited cell migration via modulating TGF-β RI/Smad and TGF-β RI/RhoA/ROCK1 pathways and suppressed lung and liver metastases in vivo. CONCLUSION: This study represents the first comprehensive exploration into the anti-metastatic potential of active fractions of PPY and PF, and their unique pennogenin glycoside 8 against EC in vitro and in vivo by modulating TGF-β RI/Smad and TGF-β RI/RhoA/ROCK1 pathways, highlighting their potential as therapeutic agents against EC metastasis.
Tian R, Li R, Quan M
… +19 more, Chen Z, Wu J, Hu H, Liu P, Peng Y, Zhao C, Song Y, Li X, Guo W, He J, Xu P, Xiao Y, Wang Y, Li R, Mao W, Yu L, Liu L, Pan H, Chen Z
ETHNOPHARMACOLOGICAL RELEVANCE: B lymphocytes are central drivers of pathogenic immune responses in autoimmune diseases and represent a promising target for therapeutic intervention. Sinomenine (SIN), a natural alkaloid...ETHNOPHARMACOLOGICAL RELEVANCE: B lymphocytes are central drivers of pathogenic immune responses in autoimmune diseases and represent a promising target for therapeutic intervention. Sinomenine (SIN), a natural alkaloid derived from Sinomenium acutum, exhibits broad immunomodulatory properties and has long been clinically used for treating autoimmune diseases, however, its direct effects on B cells and underlying mechanisms are largely unexplored. AIM OF THE STUDY: To investigate the effects of SIN on the proliferation and hyperactivation of B lymphocytes both in vitro and in vivo, and elucidate its therapeutic mechanisms in autoimmune pathologies. MATERIALS AND METHODS: Firstly, SIN was examined for its ability against lipopolysaccharides (LPS) or CpG oligodeoxynucleotide (CpG) induced B cell proliferation and hyperactivity. Then, the effects of SIN regulating B cell subsets and antibody response were studied in pristane-induced lupus and keyhole limpet hemocyanin (KLH)-immunized murine models. Furthermore, transcriptomics, small molecule-protein interaction assay (SPIA) and a series of validation experiments (including Western blotting, immunofluorescence, cellular thermal shift assay (CETSA), surface plasmon resonance (SPR), and flow cytometry) were performed to identify the key mechanisms involved. RESULTS: For the first time, our research revealed the significant inhibitory efficacy of SIN on B cell proliferation induced by LPS or CpG. Moreover, SIN significantly reduced CD69 expression, and attenuated IL-6 and IL-10 hypersecretion. Notably, SIN exerted a better inhibitory effect against CpG-mediated activation compared to LPS-induced responses. In the early immune response of pristane-induced lupus model, both SIN and prednisone significantly decreased the spleen coefficient and the proportion of CD19CD69 B cell subsets. In the KLH immunization model, SIN alleviated splenomegaly, lowered serum levels of IgG and IgM, and reduced the cell counts of B220+ cells and proportion of plasma cells (B220CD19-CD138+). Integration of transcriptomic analyses and in vitro verification revealed SIN's regulatory effects on cell cycle, the PI3K-AKT signaling pathway, along with its marked suppression of NF-κB protein activation following both LPS and CpG stimulation. In response to CpG stimulation, SIN also significantly decreased the protein levels of p-STAT3 and STAT3. Further research by SPIA identified the downregulation of interferon regulatory factor 5 (IRF5) by SIN. This effect was then corroborated by Western blot, immunofluorescence, molecular docking, CETSA, and SPR analyses, confirming SIN as a novel modulator of IRF5. Moreover, pharmacological inhibition of IRF5 by IRF5-IN-1 markedly significantly attenuated the proliferation and hyperactivation of B cells following LPS or CpG challenge, implying the critical role of IRF5 in B cells. CONCLUSIONS: Collectively, our findings suggest that SIN restrains the proliferation and hyperactivation of B lymphocytes via inhibiting IRF5 and modulating the PI3K-Akt, STAT3, and NFκB signaling cascades. These results uncover a previously unrecognized mechanism of B cell regulation and support a potential therapeutic approach for autoimmune diseases involving pathogenic B cells by SIN.
ETHNOPHARMACOLOGICAL RELEVANCE: Notoginsenoside R1 (NGR1), a primary bioactive constituent of P. notoginseng, has potent anti-inflammatory and immunomodulatory functions, but its role and mechanism in UC remain incomplet...ETHNOPHARMACOLOGICAL RELEVANCE: Notoginsenoside R1 (NGR1), a primary bioactive constituent of P. notoginseng, has potent anti-inflammatory and immunomodulatory functions, but its role and mechanism in UC remain incompletely understood and warrant further exploration. The global prevalence of ulcerative colitis (UC) is rising steadily, threatening human health. Despite advances in modern therapies, UC treatments remain limited. AIM OF THE STUDY: This work investigated whether NGR1 alleviates DSS-induced colitis by inhibiting the PGAM5-NEK7 interaction and NLRP3 inflammasome activation in macrophages, providing new mechanistic insights and therapeutic targets for UC. MATERIALS AND METHODS: UC model was established via 3% DSS administration to assess the protective efficacy of NGR1. Disease activity index (DAI), body weight and colon length were determined to assess disease severity and intestinal injury. HE staining, ELISA and immunofluorescence assays were employed to assess pathological changes in the colon and liver. Macrophages were depleted via tail vein injection of clodronate liposomes. RT-qPCR, ELISA, immunofluorescence and WB were performed to quantify macrophage markers, pro-inflammatory genes and NLRP3 inflammasome-related proteins. In vitro, LPS/ATP-stimulated bone marrow-derived macrophages (BMDMs) demonstrated that NGR1 exerts a potent inhibitory effect on NLRP3 inflammasome activation. After cell treatment, CCK-8, ELISA, RT-qPCR, and WB assays were performed to detect cell viability, inflammatory cytokine levels, and the expression of related genes and proteins. Meanwhile, molecular docking was utilized to verify the interactions among PGAM5, NEK7 and NLRP3. Co-IP in 293T cells overexpressing PGAM5, NEK7 and NLRP3 further verified their interplay. RESULTS: Compared with the Model group, NGR1 intervention markedly reduced the DAI scores, ameliorated body weight loss and colon shortening, alleviated colonic and hepatic pathological damage, upregulated the expression of intestinal barrier tight junction proteins (ZO-1, Occludin and Claudin-1), and decreased LPS levels and the release of TNF-α, IL-1β and IL-6 in serum and tissues. Macrophage depletion was observed to markedly abolish protective effects of NGR1, and reduced PGAM5, NEK7, NLRP3, Caspase-1, C-Caspase-1 levels. In vitro, NGR1 inhibited inflammatory factor secretion and NLRP3 inflammasome activation in LPS/ATP-stimulated BMDMs, these effects were partially blocked after PGAM5 or NEK7 gene silencing. Co-IP assays confirmed endogenous interactions between PGAM5, NEK7 and NLRP3. And the molecular docking showed NGR1 could bind to PGAM5, and NGR1 intervention significantly suppressed the expression of the ternary complex formed by the three proteins. CONCLUSION: This study revealed that NGR1 disrupted the PGAM5-NEK7 interaction, restrained NLRP3 inflammasome assembly in macrophages, and relieved colonic damage triggered by DSS, providing a novel target for UC treatment.
ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal plants remain an important source of therapeutic agents in traditional medicine. Umbilicus horizontalis (Guss.) DC. (U. horizontalis) is traditionally used in Mediterranean folk...ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal plants remain an important source of therapeutic agents in traditional medicine. Umbilicus horizontalis (Guss.) DC. (U. horizontalis) is traditionally used in Mediterranean folk medicine as a cicatrizing agent for wound healing and for the treatment of inflammatory conditions and pain. However, its phytochemical composition and pharmacological properties remain largely unexplored. AIM OF THE STUDY: This study aimed to investigate the phytochemical composition of aerial parts of U. horizontalis and to evaluate their antioxidant, analgesic, anti-inflammatory, and neuroprotective activities in order to provide scientific evidence supporting its traditional uses. MATERIALS AND METHODS: Leaves and stems were extracted and fractionated using different solvents. Phytochemical characterization was performed using total phenolic and flavonoid content determination and GC-MS analysis. Antioxidant activity was evaluated using DPPH scavenging, ABTS scavenging, reducing power, and phenanthroline assays. Analgesic activity was assessed using the acetic acid-induced writhing test in mice, while anti-inflammatory activity was evaluated using the xylene-induced ear edema model. Neuroprotective potential was investigated through acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition assays. RESULTS: Stem crude extract (SCrE) showed the highest phenolic (209.37 ± 7.98 μg GAE/mg) and flavonoid (87.42 ± 1.72 μg QE/mg) contents. GC-MS analysis revealed the presence of several terpenoid-related constituents. Antioxidant activity varied according to plant part and solvent, with leaf ethyl acetate extract (LEAE) exhibiting the strongest DPPH scavenging activity (IC = 6.46 ± 0.22 μg/mL), while stem aqueous extract (SAqE) was most active in the ABTS scavenging assay (IC = 4.67 ± 0.84 μg/mL). The SCrE also demonstrated strong reducing power and phenanthroline activity (CA. = 12.67 ± 0.91 and 4.96 ± 0.28 μg/mL, respectively). In vivo assays revealed significant analgesic activity in the acetic acid-induced writhing model, with SCrE producing 87.37% inhibition at 500 mg/kg. LCrE also showed marked anti-inflammatory activity, completely inhibiting xylene-induced ear edema (100% inhibition) at 250 mg/kg. Furthermore, SCrE exhibited AChE inhibitory activity (IC = 33.72 ± 1.99 μg/mL), approaching the activity of galantamine (p = 0.9945), while SEAE showed strong BChE inhibition (IC = 3.14 ± 0.55 μg/mL), significantly higher than galantamine (p < 0.0001). Correlation analysis suggested that phenolic compounds mainly contributed to antioxidant activity, while other metabolites likely underlie enzyme inhibition. CONCLUSION: The pronounced biological activities of U. horizontalis are likely attributed to its high polyphenol and flavonoid content, supporting its traditional medicinal uses. Future research should focus on isolating the active compounds, elucidating their molecular mechanisms, and assessing safety and pharmacokinetics to explore potential therapeutic applications.
ETHNOPHARMACOLOGICAL RELEVANCE: Lindera aggregata (Kosterm.) is a traditional Chinese medicine used for soothing the liver and regulating qi, as documented in classic formulas such as Tiantai Wuyao San and Simo Decoction...ETHNOPHARMACOLOGICAL RELEVANCE: Lindera aggregata (Kosterm.) is a traditional Chinese medicine used for soothing the liver and regulating qi, as documented in classic formulas such as Tiantai Wuyao San and Simo Decoction. However, the active compounds and molecular mechanisms underlying its hepatoprotective effects remain poorly understood. AIM OF THE STUDY: This study aimed to evaluate the therapeutic potential of linderane (LDR), a furanosesquiterpene from L. aggregata, in metabolic dysfunction-associated steatotic liver disease (MASLD) and to explore its mechanisms, with a focus on STAT3-mediated immunometabolic pathways. MATERIALS AND METHODS: MASLD was induced in C57BL/6J mice by feeding a high-fat diet (HFD). LDR was administered orally, and metabolic, inflammatory and histological parameters were assessed. Mechanistic studies were performed in primary hepatocytes and macrophages using Western blotting, RT-qPCR, immunofluorescence, transcriptomics, and cellular thermal shift assays (CETSA). RESULTS: LDR significantly improved fasting and postprandial glucose levels, enhanced insulin sensitivity, and restored GLUT2/GLUT4 expression. It activated the IRS2/Akt/FoxO1 signaling axis, suppressed gluconeogenic gene expression, and improved hepatic lipid metabolism via modulation of SREBP-1c and PPARα/γ. In macrophages, LDR inhibited M1 polarization and pro-inflammatory cytokine production by suppressing STAT3 phosphorylation, without affecting JAK2 expression. CETSA confirmed a direct interaction between LDR and STAT3 protein. CONCLUSION: This study demonstrates that STAT3 activation suppresses IRS2-FoxO1 signaling. Linderane targets STAT3 to relieve this suppression while also directly activating the IRS2-FoxO1 axis, thereby ameliorating MASLD. These findings support linderane as a promising natural agent that targets STAT3 and modulates immunometabolic pathways to alleviate MASLD.