Neuroinflammation is a key driver and an important therapeutic target of neurodegeneration. Dysregulated nuclear factor-κB (NF-κB) signaling plays a central role in neuroinflammation and depends on M1 and K63 ubiquitination of NF-κB essential modulator (NEMO). Ivangustin (IVA) is a sesquiterpene lactone isolated from Inula japonica Thunb., a traditional Chinese medicine has been used to treat inflammatory diseases. But its anti-neuroinflammatory effects and molecular targets remain unclear. This study aimed to evaluate the anti-neuroinflammatory effects of IVA and elucidate its direct functional target and underlying mechanism. Integrated in vitro, in vivo, and in silico/bioinformatics approaches were employed. The anti-neuroinflammatory activity of IVA was evaluated in LPS-stimulated BV2 cells, primary microglia, and an LPS-induced acute cognitive impairment mouse model. Mechanistic studies were performed using pull-down, liquid chromatography-tandem mass spectrometry (LC–MS/MS), cellular thermal shift assay (CETSA), drug affinity responsive target stability (DARTS), gene knockdown, point mutation and bioinformatics analyses. In vitro, IVA significantly inhibited nitric oxide (NO) production and mRNA and protein expression of pro-inflammatory factors in LPS-stimulated microglia. Furthermore, IVA significantly suppressed inhibitor of NF-κB kinase α/β (IKKα/β) and inhibitory subunit of NF-κB alpha (IκBα) phosphorylation, IκBα degradation, and NF-κB p65 phosphorylation and nuclear translocation, thereby blocking the expression of NF-κB-targeted genes. Its anti-neuroinflammatory activity was attenuated by thiol donors, indicating the importance of its electrophilic lactone moiety. Furthermore, ubiquitin conjugating enzyme E2L3 (UBE2L3) was identified as the functional target of IVA in BV2 cells, whereas UBE2L3 knockdown markedly attenuated IVA-mediated anti-neuroinflammatory action. Bioinformatics analysis suggests UBE2L3 overexpression as a potential risk factor for neurodegenerative diseases. Mechanistically, IVA covalently bound to the catalytic cysteine 86 of UBE2L3, thereby inhibiting UBE2L3-mediated M1 and K63 ubiquitination of NEMO, which is crucial for NF-κB activation. In vivo, IVA ameliorated LPS-induced cognitive impairment and neuroinflammation in mice via inhibiting Iba1 expression and suppressing NF-κB activation. IVA exerts anti-neuroinflammatory effects by covalently targeting UBE2L3 and suppressing NF-κB signaling. These findings also identify UBE2L3 as a potential therapeutic target for neuroinflammation and support IVA as a promising lead compound for further preclinical investigation of neuroinflammatory disorders.
Traditional Chinese Medicine (TCM) clinical practice depends on both codified theoretical knowledge and practitioner-specific experience, which differ in their data sources, reasoning patterns, and scope of generalization. We developed DFGLM-TCM, a modular large language model-based service system that separately models these two knowledge types through task-oriented components and coordinates them within a unified multi-task architecture. The knowledge-oriented component integrated a curated 22-GB TCM corpus containing approximately 3 million structured entries, a manually validated knowledge graph with more than 200,000 entities, and retrieval-augmented generation for literature retrieval and general TCM question answering. The experience-oriented component was trained using approximately 5000 authentic outpatient records from a senior TCM practitioner and 2000 expert-reviewed augmented cases to provide practitioner-specific diagnostic and prescription references. Through standardized interfaces and role-based access, the system supports knowledge retrieval, question answering, structured consultation, and prescription reference for clinicians, patients, and students. In an expert-rated evaluation of 100 TCM knowledge questions, DFGLM-TCM achieved the highest descriptive mean score among the evaluated models (4.48 ± 0.76). In 454 independent pulmonary-nodule cases, the experience-oriented component achieved a prescription-consistency score of 8.95 ± 0.63, exceeding that of the model additionally trained with general TCM knowledge (7.52 ± 0.44). A one-month assessment involving 35 physicians at three primary-care institutions suggested favorable short-term usability and acceptance. These findings highlight the value of separating general TCM knowledge from practitioner-specific experience while coordinating task-oriented training and multi-task services within a unified system, and support further evaluation of DFGLM-TCM as an auxiliary reference tool in broader clinical settings.
Abstract The genus Atractylodes (Asteraceae) is widely distributed and utilized in East and Southeast Asia, particularly in China, Japan, Korea and Thailand. In traditional Chinese medicine, representative species such as A. lancea and A. macrocephala have historically been employed to strengthen the spleen and eliminate dampness. Phytochemical investigations have led to the isolation and identification of 371 compounds from this genus, primarily encompassing polyacetylenes, terpenoids, flavonoids, phenylpropanoids, lignans, and their corresponding glycosides. In recent decades, extensive research has been conducted on the chemical constituents and pharmacological activities of plants within the genus Atractylodes . Concurrently, these crude extracts and isolated compounds have been demonstrated to exhibit diverse pharmacological activities, including anti-cancer, anti-inflammatory, gastrointestinal-regulatory, neuroprotective, hepatoprotective, and lung-protective activities. Regarding safety, their potential toxicity is closely tied to the traditional "dryness" property, which can be effectively mitigated through specialized processing methods such as bran-frying. However, rapidly accumulating data since 2021 has created a critical gap, rendering previous reviews insufficient to reflect the current research landscape. To address these emerging updates, this review systematically organizes recent advances in the traditional uses, phytochemistry, pharmacology, and safety profiles of the genus Atractylodes . Crucially, a critical analysis of structure–activity relationships is integrated to provide concrete, structured guidelines for future mechanistic exploration and rational drug development.
Abstract Background Remote electroacupuncture (EA) conditioning on forearm protects against myocardial ischemia–reperfusion injury (MIRI). However, the cardioprotective mechanisms of RIC have not been fully elucidated. This study aimed to investigate whether EA could balance the pro-survival and pro-inflammatory responses in MIRI rats via SDF-1α/CXCR4 axis. Methods SD rats were subjected to MIRI and underwent EA treatment before reperfusion therapy. Infarct size, cardiac function, and neutrophil infiltration were evaluated after 24-h of reperfusion. The mRNA and protein levels of SDF-1α in the myocardial border and infarct zones, were measured by RT-qPCR and Western blotting to reveal its spatial distribution. We examined serum SDF-1α levels and its myocardial scavenger receptor CXCR7 expression to elucidate the mechanism underlying its spatial distribution. Then, CXCR4/ERK pathway was evaluated at 15 min and 24 h after EA to examine the time-dependent dual effects of SDF-1α on CXCR4. Results Our results showed that EA attenuated infarct size and mortality, improved cardiac function after 24 h of MIRI. We identify that SDF-1α is a key humoral factor mediating EA-induced cardioprotection. In ischemic myocardium, the infarct zone exhibited a higher SDF-1α concentration than the border zone following EA treatment, a difference potentially linked to the elevated expression of CXCR7 in the border zone. In the early phase of reperfusion (15 min), our study showed that EA promoted the expression of the CXCR4 receptor, which in turn triggered ERK activation. This rapid ERK activation via SDF-1α/CXCR4 promoted cardiomyocyte survival and attenuated reperfusion-induced apoptosis. While at 24 h post-reperfusion, saturating signals of SDF-1α by EA downregulated CXCR4 expression, along with phosphorylation of its downstream effectors ERK and NF-κB p65. This coordinated downregulation facilitates inflammatory resolution by promoting neutrophil apoptosis. Conclusions Cardioprotection by EA can be mediated by humoral factors SDF-1α that accumulate in infarct myocardium. SDF-1α triggers pro-survival CXCR4/ERK signaling at 15 min of reperfusion, whereas it promotes neutrophil apoptosis via CXCR4 degradation to facilitate inflammatory resolution by 24 h, thereby fine-tuning the dual role of the SDF-1α/CXCR4 axis in MIRI.
Traumatic brain injury (TBI) is associated with high morbidity, disability, and long-term neurological sequelae. Mitochondrial dysfunction is a central component of secondary injury after TBI, contributing to impaired energy metabolism, oxidative stress, calcium dysregulation, neuroinflammation, and neuronal apoptosis. This review summarizes current evidence regarding acupuncture-mediated regulation of mitochondrial homeostasis after TBI, focusing on structural homeostasis, quantitative homeostasis, and functional/metabolic homeostasis. We distinguish direct evidence from TBI models, indirect evidence from related brain injury models, and hypothesis-generating mechanisms. Particular attention is given to mitochondrial dynamics, mitochondrial biogenesis, mitophagy, energy metabolism, oxidative stress, calcium signaling, mitochondrial membrane potential, apoptosis, intercellular mitochondrial transfer, and putative upstream neural and humoral pathways. Current preclinical evidence suggests that acupuncture, particularly electroacupuncture, may influence mitochondrial homeostasis after TBI through multidimensional and context-dependent mechanisms. However, direct TBI-specific evidence remains limited, and several proposed mechanisms require further validation before acupuncture-mediated mitochondrial regulation can be considered a clinically established therapeutic strategy.
Yiqi Huoxue Jiedu Formula (YHJF) is a traditional Chinese medicine formula that has been used as an adjunctive therapy for sepsis for nearly two decades. Previous clinical studies showed that YHJF improves Sequential Organ Failure Assessment (SOFA) scores and modulates gut microbiota in elderly patients with pneumonia-associated sepsis. However, the mechanism by which YHJF protects against sepsis-associated acute lung injury (SALI) remains unclear. A murine SALI model was established by cecal ligation and puncture (CLP). Therapeutic effects were evaluated by histopathology, micro-CT, pulmonary function assessment, and ELISA. Mechanistic studies included proteomic analysis of lung tissues and LPS-stimulated MH-S macrophages, pharmacological modulation with Mdivi-1 and urolithin A (UA), macrophage-epithelial co-culture, HPLC fingerprinting, UPLC-HRMS, and molecular docking. YHJF significantly improved 7-day survival and ameliorated lung injury, pulmonary edema, respiratory dysfunction, and systemic inflammation in mice with CLP-induced SALI. Proteomic profiling and subsequent functional assays suggested that enhanced mitophagy in macrophages represents a central protective mechanism. In vivo, YHJF increased autophagosome formation and PINK1/Parkin co-localization in BALF-derived alveolar macrophages. In vitro, YHJF restored mitochondrial homeostasis by activating PINK1/Parkin-dependent mitophagy in macrophages. This was accompanied by reduced cytoplasmic mtDNA leakage, downregulated cGAS expression, and suppression of the STING–TBK1–IRF3 pathway and subsequent type I interferon responses. Pharmacological inhibition of mitophagy with Mdivi-1 abolished these protective effects of YHJF, whereas activation with UA augmented them, demonstrating that mitophagy is necessary for YHJF-mediated protection. In a macrophage–epithelial co-culture system, YHJF-treated macrophages alleviated LPS-induced apoptosis in MLE-12 alveolar epithelial cells. Furthermore, chemical analysis integrated with molecular docking identified aloe-emodin, rhein, and genistein as candidate bioactive constituents of YHJF that likely contribute to its regulation of macrophage mitophagy. YHJF protects against SALI by restoring macrophage mitophagy and suppressing mtDNA-STING-mediated inflammatory signalling. These findings support YHJF as a potential therapeutic strategy for sepsis-associated lung injury.
Abstract Diabetic wounds represent one of the most intractable complications of diabetes, arising from a highly complex and self-perpetuating pathological microenvironment. This milieu is characterized by chronic inflammation, oxidative stress, impaired angiogenesis, peripheral neuropathy, immune dysregulation, recurrent infection, and marked dysfunction of resident cells and the extracellular matrix (ECM). These intertwined abnormalities not only hinder wound closure but also significantly increase the risk of chronic infection, amputation, and disability, thereby imposing substantial clinical and socioeconomic burdens. Conventional therapies, which typically target isolated aspects of wound pathology, often fail to disrupt the vicious cycle of non-healing, underscoring the urgent need for integrated, multi-target therapeutic strategies. Chinese herbal medicine (CHM) encapsulates a holistic therapeutic paradigm that is intrinsically aligned with the multi-factorial nature of diabetic wound management. CHM offers a diverse array of bioactive ingredients, including polysaccharides, saponins, flavonoids, alkaloids, and phenolics, that collectively exert anti-inflammatory, antioxidant, pro-angiogenic, neuroprotective, immunomodulatory, antimicrobial, and pro-proliferative effects. Moreover, many of these ingredients exhibit pleiotropic effects, enabling concomitant modulation of multiple pathological pathways to address the inherent complexity of diabetic wounds. However, the clinical translation of raw CHM is frequently hindered by poor bioavailability and rapid degradation at the wound site. Recent advances in CHM-based innovative delivery platforms have provided a transformative framework to overcome these limitations. This review highlights the emergence of CHM-loaded hydrogels and CHM-derived plant exosome-like nanovesicles (PELNs) as next-generation interventions. CHM-loaded hydrogels act as biomimetic scaffolds that facilitate controlled release and adaptively respond to microenvironmental cues (e.g., pH and reactive oxygen species (ROS)). Simultaneously, CHM-derived exosome-like nanovesicles serve as natural, biocompatible nanocarriers that enhance cellular uptake and molecular signaling, promoting regenerative healing through intercellular communication pathways. In this review, we systematically summarize the pathological features of diabetic wounds and synthesize the modern pharmacological mechanisms of representative CHM ingredients. We further evaluate the synergistic integration of CHM with advanced biomaterials and nanotechnology, emphasizing their translational potential from preclinical models to clinical evidence. By bridging traditional wisdom with cutting-edge bioengineering, these platforms offer a robust evidence-based framework for the modernized management of diabetic wounds.
Osteoporosis is increasingly linked to metabolic dysregulation. Medicine–food homologous (MFH) materials contain diverse natural bioactives with reported osteoprotective effects, but their overall evidence landscape remains insufficiently integrated. To synthesize preclinical evidence on MFH-derived bioactives that improve osteoporosis-related phenotypes and to discuss their possible trans-organ actions under metabolic disturbance. PubMed, Web of Science, Scopus, and CNKI were searched from January 2000 to September 2025. Original in vivo or in vitro studies were included when a defined MFH-derived constituent or standardized single-material extract was tested in an osteoporosis-relevant model and reported both bone-related and metabolism-related outcomes. A total of 38 bioactive components from 24 MFH materials were identified and regrouped into seven higher-level natural-product categories, including flavonoids, phenolic and polyphenolic compounds, polysaccharides, saponins, terpenoids, proteins/peptides, and other specialized metabolites. Across studies, osteoprotective effects were frequently accompanied by parallel improvements in lipid metabolism, inflammatory status, oxidative stress, gut microbiota, or related metabolites. These findings suggest that MFH bioactives may act beyond bone-local signaling alone. In the discussion, this pattern was further interpreted through liver-bone and gut-bone. MFH-derived bioactives show potential to improve osteoporosis-related phenotypes and may exert broader systemic regulatory effects, although specific mediators and causal links still require validation.
Inflammatory bowel disease (IBD) is a relapsing inflammatory disorder of the gastrointestinal tract with increasing global incidence. Current therapies are often limited by side effects, loss of efficacy, and high cost, underscoring the need for safer and more effective alternatives, particularly multi-target agents derived from natural products. This study aimed to elucidate the protective mechanisms of Yiyi Fuzi Baijiang formula (YFB), a traditional Chinese medicine (TCM) formulation, against dextran sulfate sodium (DSS)-induced acute colitis, focusing on its systemic regulation of the gut barrier–microbiota–metabolism axis. We employed an integrated approach combining network pharmacology, UPLC-Q-TOF-MS/MS-based phytochemical analysis, in vivo evaluation in a DSS-induced colitis mouse model, 16S rRNA gene sequencing, and untargeted metabolomics to assess the effects of YFB and uncover its mechanisms of action. Network pharmacology predicted, and experiments confirmed, that core YFB components (e.g., quercetin, kaempferol) act via IL-17, TNF, and NF-κB pathways. YFB administration dose-dependently improved disease activity index, colon shortening, and histopathology in colitis mice. It restored intestinal barrier integrity by upregulating ZO-1, Occludin, and MUC2, while suppressing pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-17A) and NF-κB activation. Critically, YFB promoted epithelial repair by restoring the expression of intestinal stem cell marker LGR5 and progenitor cell marker SOX9, and by normalizing the aberrant increase in endocrine cell marker CHGA. YFB treatment was associated with reversal of DSS-induced gut microbiota dysbiosis, restoration of diversity, enrichment of beneficial bacteria (e.g., Lachnospiraceae), and suppression of opportunistic pathogens (e.g., Enterobacteriaceae). Untargeted metabolomics showed that YFB treatment was associated with modulation of DSS-altered fecal metabolites (e.g., fatty acids, bile acids) and pathways such as "microbial metabolism in diverse environments". YFB, when administered concomitantly with DSS, protects against DSS-induced colitis via the synergistic effects of its multi-component system. Its mechanism entails systemic regulation of the gut barrier–microbiota–metabolism axis, involving suppression of NF-κB-driven inflammation, promotion of intestinal epithelial repair (via LGR5/SOX9/CHGA modulation), restoration of the intestinal barrier, alterations in gut microbiota, and modulation of host–microbial co-metabolism. These findings provide a scientific basis for YFB's clinical application and highlight the value of TCM formulations in managing complex multi-factorial diseases.
Diabetic retinopathy (DR) is a leading cause of blindness. While anti-vascular endothelial growth factor (VEGF) therapy is effective, its utility is limited by variable patient response and the need for frequent injections. Therefore, identifying new therapeutic targets for DR is imperative. Emerging evidence indicates that astrocytes contribute to endothelial dysfunction in DR, suggesting that targeting astrocyte-endothelial cell crosstalk represents a promising therapeutic strategy. To evaluate the therapeutic potential of Salvianolic acid A (Sal A) for DR, elucidate the molecular mechanisms by which it modulates astrocyte-endothelial cell interactions, and develop a liposome-based nanodelivery system to enhance its efficacy. The protective effects and mechanisms of Sal A were systematically investigated using a streptozotocin (STZ)-induced diabetic mouse model, complemented by a suite of in vitro and molecular approaches including co-culture models, transcriptomic analysis, and target validation assays. Retinal vascular structure and barrier function were assessed in vivo via immunofluorescence staining and Evans Blue leakage assays. Endothelial cell behaviors were examined in vitro using wound healing, Transwell migration, tube formation, and spheroid sprouting assays. Transcriptomic profiling was performed by RNA sequencing (RNA-seq). The direct target of Sal A was identified and validated using MS-based drug-affinity responsive target stability (DARTS) screening, cellular thermal shift assay (CETSA), and microscale thermophoresis (MST). Expression of key signaling molecules was measured by western blotting, enzyme-linked immunosorbent assay (ELISA), and quantitative real-time PCR (qRT-PCR). Liposome@Sal A was prepared and characterized for its physicochemical properties (dynamic light scattering, transmission electron microscopy), stability, and therapeutic efficacy in vitro and in vivo. Sal A treatment ameliorated retinal vascular abnormalities in diabetic mice, evidenced by increased VE-cadherin and NG2 expression, decreased α-smooth muscle actin (α-SMA) expression, and reduced acellular capillary formation, collectively restoring vascular integrity. Mechanistically, astrocyte-derived soluble Semaphorin 4D (sSema4D) promoted endothelial hyperactivation via the PlexinB1/RhoA/ROCK/pMLC2 signaling cascade. Sal A directly bound to the Arg92 residue of membrane-bound Sema4D on astrocytes, significantly inhibiting sSema4D shedding and its subsequent deleterious effects on endothelial cells. Furthermore, Liposome@Sal A enhanced retinal drug delivery and demonstrated superior therapeutic efficacy compared to free Sal A in diabetic mice. Sal A preserves retinal vascular structure and function in DR by binding to astrocytic Sema4D at Arg92, thereby inhibiting sSema4D shedding and downstream PlexinB1/RhoA/ROCK/pMLC2 signaling, which modulates astrocyte-endothelial cell crosstalk. Liposomal encapsulation significantly potentiates the therapeutic efficacy of Sal A, positioning it as a promising drug candidate for DR treatment.
Heart failure with preserved ejection fraction (HFpEF) has currently emerged as a predominant and challenging subtype of heart failure, with high morbidity and mortality. However, the efficacy of current therapeutic strategies for HFpEF remains unsatisfactory. The traditional Chinese medicine formulation Shenfu Qiangxin pill (SFQX) ameliorates clinical symptoms in patients with heart failure, but its precise mechanisms for HFpEF remain to be elucidated. This study aimed to investigate the therapeutic potential of SFQX for HFpEF and to elucidate the mechanisms underlying its effects. UPLC-Q-TOF-MS/MS analysis was performed to identify the major active ingredients of SFQX. The HFpEF mouse model was established using a high-fat diet and an Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME) to evaluate the therapeutic efficacy of SFQX. We employed an integrated approach combining single-cell RNA sequencing (scRNA-seq) with functional and molecular validation to characterize SFQX-induced changes in cardiac cellular composition, cell-state remodeling, and tissue-level phenotypes in HFpEF. We show that SFQX exerts therapeutic effects against HFpEF by coordinately modulating maladaptive cardiac cell subsets. Specifically, SFQX was associated with reprogramming of pathogenic immune cell polarization, normalization of fibroblast state heterogeneity, enhanced endothelial metabolic adaptability, and restoration of lymphatic endothelial homeostasis. These multicellular changes were accompanied by improved cardiac structure and function, reduced fibrosis and inflammation, enhanced lymphatic drainage capacity, and alleviated myocardial edema. Our findings highlight the ability of SFQX, as a multicomponent agent, to precisely regulate the highly heterogeneous pathology of HFpEF at a network level. This work not only establishes a mechanistic link between holistic principles of traditional medicine and modern biology but also provides a novel theoretical basis for SFQX's efficacy in multifactorial diseases.
The host microbiota and hepatic drug-metabolizing enzymes are important mediators of the metabolism and biological effects of herbal components. Through bidirectional interactions, herbal medicines can also reshape the host microbial community. The clinical efficacy of Wendan Decoction (WDD) in treating metabolic dysfunction-associated fatty liver disease (MAFLD) has been well established. However, its interactions with the host microbiota through the gut-liver axis remain unclear. This study aimed to investigate the mechanism by which WDD modulates host microbial activity through the gut-liver axis to ameliorate MAFLD. MAFLD models were established by high-fat diet (HFD) feeding and subsequently treated with WDD, Parasutterella excrementihominis (P. excrementihominis), or 7α-OH-T. The ABX group underwent antibiotic-mediated microbiota depletion before treatment. Multi-omics analyses were used to characterize the dynamic trajectories of microbiota-derived metabolites. These analyses included targeted bile acid (BA) profiling of serum, 16S rRNA gene sequencing and untargeted metabolomics of cecal contents, and proteomics and untargeted metabolomics of liver tissue. Hematoxylin and eosin, Oil Red O, and Alcian blue-periodic acid-Schiff staining were used to assess pathological changes in the liver and intestinal tissues during MAFLD. ELISA, Western blotting, and other assays were performed to quantify markers of inflammation and lipid metabolism. Following UPLC/UV detection of 7α-OH-T in portal vein serum, molecular docking and molecular dynamics simulations, together with cellular thermal shift assays (CETSA) and microscale thermophoresis (MST), were used to validate FXR as a target of 7α-OH-T. WDD alleviated hepatic steatosis, intestinal inflammation, and barrier dysfunction in MAFLD, but these effects depended on the integrity of the host microbiota. 16S rRNA gene sequencing showed that WDD promoted the growth of beneficial bacteria, including Bacteroides and Parasutterella. Combined analysis of targeted serum BA metabolomics and untargeted metabolomics of cecal contents indicated that WDD-mediated modulation of the host microbiota reduced the total serum BA load, increased alternative-pathway metabolites, including CDCA and TCDCA, in the liver and intestine, and decreased toxic secondary BAs, including DCA and LCA. Steroid and fatty acid metabolites, such as 7α-OH-T, were also increased. Pearson correlation analysis and P. excrementihominis transplantation experiments suggested that the increase in 7α-OH-T was closely associated with P. excrementihominis. Untargeted liver metabolomics and serological analyses confirmed that gut-derived 7α-OH-T entered the liver through the portal vein and acted on hepatic targets via the gut-liver axis. In animal experiments involving exogenous 7α-OH-T supplementation and in MAFLD THLE-2 cell models treated with 7α-OH-T, 7α-OH-T ameliorated hepatic lipid accumulation and promoted lipid utilization in THLE-2 cells. A series of interaction assays, including CETSA and MST, identified FXR as a target of 7α-OH-T. Furthermore, 7α-OH-T markedly activated the FXR/PPARα/CYP4A12A axis and served as a key messenger through which WDD-mediated regulation of Parasutterella alleviated MAFLD via the gut-liver axis. WDD increased the abundance of P. excrementihominis and the level of the potentially associated metabolite 7α-OH-T. Through the portal circulation, 7α-OH-T promoted gut-liver crosstalk and targeted the FXR/PPARα/CYP4A12A axis, thereby ameliorating MAFLD.
Acupuncture prescriptions involve complex compatibility mechanisms grounded in multi-symptom and multi-acupoint interactions, embodying millennia of clinical experience. Despite growing interest in computational acupoint recommendations, significant challenges persist due to sparse clinical data and the insufficient modelling of symptom-acupoint relationships, posing considerable hurdles to effective prediction. We introduced an acupoint compatibility prediction framework with graph neural networks (GNN) and fine-tuned bidirectional encoder representations from transformers (termed GNN-BERT-Attention). The heterogeneous feature interaction learning mechanism was introduced to model symptom-acupoint interactions through heterogeneous graph construction, capturing semantic features and relational patterns in a unified space, which alleviated the sparsity of data. Neural collaborative filtering is utilised via label-aware fusion to iteratively refine the confidence of predictions, while Focal Loss and randomised augmentation strategies enhance robustness against imbalanced label distribution. Comprehensive experiments demonstrate the superiority of the proposed GNN-BERT-Attention model over State-of-the-Art (SOTA) baselines in precision, recall, ranking-based metrics and robustness. Ablation studies validate the effectiveness of each architectural module, and hyperparameter tuning confirms models’ stability. A web-based demonstration system further validates clinical applicability, enabling real-time, interpretable acupoint recommendations. This study contributes to enhancing the performance of acupoint prediction, ultimately benefiting the efficiency and precision of acupuncture treatment while providing a theoretical foundation for optimising prescriptions and advancing evidence-based traditional Chinese medicine interventions.
Scutellariae Radix-Coptidis Rhizoma herb pair is a classic traditional Chinese medicine (TCM) combination with heat-clearing, dampness-drying and detoxifying effects, extensively applied clinically against bacterial infections. Traditional water decoction and ethanol pretreatment are mainstream extraction approaches, whereas existing comparisons merely emphasize quantitative variations of bioactive compounds, neglecting the underlying supramolecular assembly differences of effective substances. Comparative analysis of solvent-induced supramolecular variations provides a novel perspective to interpret differential pharmacodynamic material bases of TCM formulas. Macroscopic and microscopic morphologies of HH-0 and HH-50 were firstly characterized by visual inspection, SEM and DLS. Their phytochemical profiles were subsequently analyzed by UHPLC-Q-Orbitrap HRMS and HPLC. Antibacterial activities against MRSA were further quantified via turbidimetry and plate assays. UV, FT-IR and MD simulations revealed divergent supramolecular assembly behaviors. Finally, untargeted metabolomics and assembly-peptidoglycan interaction simulations elucidated the underlying anti-MRSA mechanisms. The solvent pretreatment endowed HH-0 and HH-50 supramolecules with distinct characteristics. HH-50 featured rougher supramolecular surfaces and higher Zeta potential with favorable dispersibility. The two supramolecules shared consistent active constituents, and HH-50 showed elevated berberine abundance and enhanced anti-MRSA activity. Notably, HH-50 maintained superior antibacterial efficacy and interference against peptidoglycan synthesis and energy metabolism even at identical berberine doses. This confirmed that divergent supramolecular conformations determined antibacterial differences, as diminished hydrogen bonds and weakened van der Waals and electrostatic interactions enhanced the binding affinity between assemblies and MRSA. Beyond chemical composition, solvent pretreatment (water versus 50
Metabolic dysfunction-associated steatohepatitis (MASH), the progressive form of metabolic dysfunction-associated fatty liver disease (MAFLD), is tightly linked to gut microbiota dysbiosis and disrupted bile acid (BA) homeostasis. Floridoside (Flor), a marine glycoside from the edible seaweed Pyropia haitanensis (P. haitanensis), exerts promising biological activities. However, protocols for its high-purity preparation and the mechanisms underlying its anti-MASH effects remain unclear. To develop a protocol for the preparation of high-purity Flor and its isomer isofloridoside (Isoflor) from P. haitanensis, and to elucidate how Flor alleviates MASH via regulating gut microbiota and BA metabolism. High-purity Flor and Isoflor were isolated via integrated chromatography, with their chemical structures confirmed by LC–MS and NMR. Anti-MASH efficacy was evaluated in a high-fat diet (HFD)-induced murine MASH model. The underlying mechanisms were explored using multi-omics analyses, including transcriptomics, gut microbiota metagenomics and BA-targeted metabolomics, and further validated by molecular docking, molecular dynamics simulation and western blotting; the compounds’ biosafety was evaluated using zebrafish. High-purity Flor and Isoflor were successfully isolated, each with a purity of ≥ 99.0
Diabetic nephropathy (DN) is a major microvascular complication of diabetes and a leading cause of end-stage renal disease. Chronic inflammation plays a pivotal role in the pathogenesis of DN. Lycorine (LY), a complex tetracyclic pyrrolo[de]phenanthridine alkaloid derived from the Amaryllidaceae family, possesses notable anti-inflammatory activity, yet its therapeutic potential in DN remains insufficiently defined. We evaluated the renoprotective effects of LY both in vivo and in vitro. Streptozotocin (STZ)-induced diabetic mice were treated with LY, and renal function and histopathological alterations were assessed. In vitro, human renal tubular epithelial HK-2 cells were exposed to high glucose plus palmitic acid (HG + PA) with or without LY. RNA-seq analysis was performed to identify LY-regulated pathways. Molecular docking, surface plasmon resonance (SPR) assay, and cellular thermal shift assay (CETSA) were used to evaluate the interaction between LY and receptor for advanced glycation end products (RAGE). RAGE siRNA-mediated knockdown was further conducted to determine whether RAGE is required for the protective effects of LY. Activation of the HMGB1/RAGE/NF-κB signaling axis and associated inflammatory mediators was analyzed by Western blotting and RT-PCR. LY markedly alleviated renal injury in STZ-induced diabetic mice, as evidenced by reduced albuminuria, improved renal function, and attenuated renal fibrosis, apoptosis, oxidative stress, and inflammation. Notably, LY did not significantly alter blood glucose levels or body weight, indicating that its renoprotective effect was independent of glycemic control. In HG + PA-treated HK-2 cells, LY significantly suppressed apotosis, oxidative stress, and inflammatory cytokine expression. Mechanistically, RNA-seq analysis identified AGE-RAGE and NF-κB signaling pathways as key pathways modulated by LY. Molecular docking, SPR, and CETSA confirmed that LY directly interacted with RAGE. Moreover, RAGE knockdown largely abolished the additional protective effects of LY, supporting RAGE as a critical molecular target. LY inhibited HMGB1/RAGE-mediated NF-κB activation, as reflected by reduced HMGB1, RAGE, p-p65, and p-IκBα levels. LY ameliorates diabetic nephropathy without affecting blood glucose levels by directly targeting RAGE and suppressing the HMGB1/RAGE/NF-κB signaling axis. These findings identify LY as a potential RAGE-targeting therapeutic candidate for inflammation-driven diabetic kidney injury.
Abstract Background Chemotherapy-induced intestinal mucositis (CIM), particularly that induced by agents such as 5-fluorouracil (5-FU), frequently leads to chemotherapy discontinuation. However, effective treatment options remain limited. Guiqi Baizhu prescription (GQBZP), a traditional Chinese medicine formula, has been reported to possess anticancer, analgesic, and anti-inflammatory activities. Purpose This study aimed to evaluate the therapeutic efficacy of GQBZP against 5-FU-induced intestinal mucositis (IM) and to clarify its underlying molecular mechanisms and material basis. Methods The protective effects and mechanistic actions of GQBZP were investigated using a murine model of 5-FU-induced IM. Potential IKKβ-targeting compounds within GQBZP were screened through virtual docking combined with CCK-8 assays and subsequently evaluated in 5-FU-stimulated human intestinal epithelial cells (HIECs) and lipopolysaccharide/interferon-γ (LPS/IFN-γ)-stimulated THP-1 macrophages. Target specificity and binding characteristics were further validated by molecular dynamics (MD) simulations, surface plasmon resonance (SPR) analysis, and experiments using IKKβ-overexpressing HEK293T cells. Results In vivo experiments demonstrated that GQBZP significantly alleviated 5-FU-induced IM by suppressing M1 macrophage polarization within intestinal tissues and restoring intestinal barrier integrity, effects closely associated with modulation of the IKKβ/NF-κB signaling pathway. Virtual screening and CCK-8 assays identified five IKKβ-targeting compounds in GQBZP: Rhamnocitrin, Toralactone, Naringenin, Liquiritigenin, and Carvacrol. These compounds markedly reduced apoptosis and pro-inflammatory cytokine production in 5-FU-treated HIECs. In addition, they inhibited M1 macrophage polarization and cytokine release in LPS/IFN-γ-stimulated THP-1 cells, accompanied by attenuation of IKKβ/NF-κB pathway activation. MD simulations, SPR assays, and functional studies in IKKβ-overexpressing HEK293T cells further confirmed that Toralactone directly binds to and suppresses IKKβ activation, whereas Rhamnocitrin modulates IKKβ activity through an indirect regulatory mechanism. Conclusion GQBZP alleviates 5-FU-induced IM by promoting recovery of the intestinal epithelial barrier and inhibiting M1 macrophage polarization through an IKKβ/NF-κB-dependent mechanism, thereby exerting synergistic anti-inflammatory effects. Toralactone was identified as a key active constituent responsible for direct inhibition of IKKβ-mediated M1 polarization. These findings suggest that targeting IKKβ to regulate macrophage polarization represents a promising therapeutic strategy for the management of CIM.
Atractylodis Macrocephalae Rhizoma (AMR) has traditionally been utilized for treating spleen deficiency diarrhea. Nevertheless, the effects and mechanisms of AMR on diarrhea caused by the consumption of cold drinks and a high-fat diet (CDHFD) remain insufficiently understood. This study aimed to explore the therapeutic effects and mechanisms of AMR in treating CDHFD-induced diarrhea. AMR was prepared as an aqueous extract, and its chemical composition was analyzed using UPLC-ESI-MS. A diarrhea model was established in ICR mice by exposure to CDHFD for four weeks, with AMR (low/high doses) administered concurrently via oral gavage. Bowel movements were evaluated using indicators such as fecal water content. Systemic inflammation was assessed by measuring pro-inflammatory cytokines via ELISA and performing peripheral blood cell counts. Intestinal barrier integrity was examined via H E, AB-PAS staining, and immunofluorescence of tight junction proteins. Gut microbiota profiling was performed using 16S rDNA sequencing. Serum lipopolysaccharide (LPS) levels were measured via ELISA to assess translocation. Finally, the regulatory effects of AMR on the A20/TRAF6/NF-κB signaling pathway were validated using Western blotting. Spearman’s correlation analysis was employed to integrate microbiota changes with host inflammatory phenotypes. AMR significantly ameliorated CDHFD-induced diarrhea. Mechanistically, AMR remodeled the gut microecology by enriching beneficial bacteria, particularly Lachnospiraceae_NK4A136 and norank_f_Muribaculaceae. Concurrently, it increased the number of goblet cells and regulated the expression of tight junction proteins to repair intestinal barrier damage and reverse hyperpermeability. The restoration of barrier function effectively blocked the systemic translocation of LPS, which subsequently inhibited the hyperactivation of the NF-κB signaling pathway, thereby reducing systemic inflammation and ultimately alleviating diarrhea. AMR exerts protective effects against CDHFD-induced diarrhea through microbiota-driven intestinal barrier restoration, which sequentially blocks the activation of the LPS/NF-κB inflammatory pathway.
