cGAS-STING pathway is a core regulator of innate immunity and inflammatory responses. Aberrant activation of the cGAS-STING pathway is closely associated with the progression of multiple inflammatory disorders. Flavonoids from Sophora tonkinensis (STF) are the main ingredients of this traditional Chinese medicine with definite anti-inflammatory activity, whereas its molecular mechanism against inflammation remains unclear. This work aimed to explore the regulatory effect and underlying molecular mechanism of STF on the cGAS-STING pathway, and verify its therapeutic efficacy on typical inflammatory diseases in vivo. BMDMs and human THP-1 were used for in vitro experiments. Western blot, RT-qPCR, ELISA, Co-IP, and Native-PAGE were performed to detect protein phosphorylation, gene expression, protein-protein interaction, and STING oligomerization. Lipopolysaccharide/D-galactosamine (LPS/D-GalN)-induced acute liver injury and LPS-induced sepsis mouse models were established to evaluate the in vivo anti-inflammatory and organ-protective effects of STF. Serum biochemical detection and gene expression analysis were carried out for in vivo sample evaluation. STF suppressed ISD-triggered activation of the cGAS-STING pathway, as evidenced by reduced phosphorylation of interferon regulatory factor 3 (IRF3) and STING, as well as decreased expression of downstream inflammatory mediators. Mechanistically, STF did not interfere with STING oligomerization but blocked the assembly of the STING-TBK1 complex and the interaction between IRF3 and TBK1. In vivo experiments demonstrated that STF dose-dependently alleviated hepatic pathological damage and hepatocyte apoptosis in a mouse model of acute liver injury. Furthermore, STF improved survival rates and attenuated multi-organ injury in septic mice by reducing the release of pro-inflammatory cytokines. STF exerts targeted anti-inflammatory activity via disrupting STING-TBK1 and TBK1-IRF3 complex assembly and inhibiting the cGAS-STING signaling cascade. It provides experimental evidence for the clinical application and translational research of Sophora tonkinensis and its flavonoid components in the treatment of cGAS-STING-related inflammatory diseases.

Alcoholic liver disease (ALD) is a multifaceted disorder in which oxidative injury, iron-dependent ferroptosis, steatosis, and inflammation are closely intertwined. Consequently, therapeutic strategies targeting a single pathway often produce limited clinical benefits. In this study, we identified polyphyllin I (PPI), a bioactive constituent isolated from Paris polyphylla, as a potent hepatoprotective agent that concurrently ameliorates these pathological phenotypes in ethanol-exposed hepatocytes and a murine ALD model. Cellular thermal shift assay (CETSA) confirmed cellular engagement between PPI and PDE6D. Mechanistically, PPI regulates PDE6D to elevate intracellular cGMP levels and subsequently activate PKG signaling. Genetic perturbation experiments revealed that PDE6D exacerbates ethanol-induced hepatocyte ferroptosis via cGMP downregulation, and the hepatoprotective effects of PPI could be recapitulated by the PDE6D degrader TMX-4100. Downstream of PKG activation, PPI simultaneously restores xCT/GPX4 expression, suppresses ACSL4-mediated lipid peroxidation to inhibit ferroptosis, promotes NRF2 nuclear translocation to strengthen antioxidant defenses, restores autophagic flux to eliminate lipid droplets, and attenuates canonical NF-κB activation to alleviate hepatic and systemic inflammation. Pharmacological inhibition of PKG abrogated all four aforementioned effects, verifying pathway dependency. These findings establish PDE6D as a druggable vulnerability in ALD and suggest that PPI simultaneously suppresses ferroptosis and reprograms metabolic and immune homeostasis through the PDE6D-cGMP-PKG axis.

Chemotherapy-induced thrombocytopenia (CIT) increases bleeding risk and compromises chemotherapy delivery. Purpurin, a hydroxyanthraquinone found in Rubia cordifolia L., has not been examined as a regulator of megakaryopoiesis. We investigated whether purpurin restores platelet production after cyclophosphamide (CTX) exposure and examined the involvement of estrogen receptor α (ESR1)-phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signalling. Purpurin was evaluated in K562 and Meg-01 megakaryocytic differentiation models, Tg(itga2b:eGFP) zebrafish, and CTX-induced thrombocytopenic mice. Megakaryocyte (MK) differentiation, DNA ploidy, cytoskeletal remodelling, platelet recovery, and haemostatic function were assessed. Computational target prediction was used for hypothesis generation, followed by drug affinity responsive target stability analysis and pharmacological inhibition of ESR1 and PI3K. At concentrations that did not cause detectable cytotoxicity (2.5-10 μM), purpurin increased CD41[+]CD42b[+] cells, MK ploidy, F-actin remodelling, and expression of NF-E2, GATA1, RUNX1, and FOSB. Purpurin increased thrombocyte abundance in zebrafish and restored platelet counts, MK maturation, and haemostatic function in CTX-treated mice. Purpurin increased the resistance of ESR1 to proteolysis in the drug affinity responsive target stability (DARTS) assay. Pharmacological inhibition of ESR1 or PI3K attenuated purpurin-induced differentiation and PI3K/AKT phosphorylation. These findings support the involvement of ESR1-associated PI3K/AKT signalling through which purpurin promotes megakaryopoiesis and platelet recovery. Purpurin warrants further evaluation as a natural-product lead for CIT, including genetic target validation, pharmacokinetic characterization, and assessment in tumour-bearing models.

Dihydroartemisinin (DHA), a bioactive derivative of Artemisia annua L. (AAL), has shown antitumor activity in several malignancies, including non-small cell lung cancer (NSCLC). However, the molecular determinants linking DHA exposure to ferroptosis in NSCLC remain incompletely understood. This study aimed to identify functionally relevant targets of DHA-associated ferroptotic stress and to investigate the role of NAD(P)H quinone dehydrogenase 1 (NQO1) in this process. An integrated network pharmacology and transcriptomic approach was used to identify ferroptosis-related candidate targets of A. annua. Molecular docking and molecular dynamics simulations were used for target prioritization, followed by surface plasmon resonance (SPR) and cellular thermal shift assay (CETSA) to evaluate the interaction between DHA and NQO1. The effects of DHA on ferroptosis-related phenotypes and redox homeostasis were examined in A549 and PC9 cells. NQO1 overexpression, NQO1 knockdown, and GPX4 rescue experiments were performed to assess pathway involvement. Antitumor activity and NQO1-associated effects were further evaluated in mouse tumor models. Network pharmacology identified 15 active components of AAL and 61 NSCLC-associated therapeutic targets enriched in ROS-related and ferroptosis-associated pathways. Among 10 ferroptosis-related core targets, NQO1 was prioritized based on disease relevance, predicted DHA interaction, selective protein suppression, and complementary computational and experimental evidence. DHA was selected for mechanistic validation based on its pharmacological relevance and anti-NSCLC activity. SPR supported direct DHA-NQO1 binding (KD = 7.61 × 10[-6] M). Mechanistically, DHA suppressed NQO1 expression through transcriptional inhibition and enhanced protein degradation. DHA induced ferroptosis hallmarks, including ROS burst, LPO and Fe[2+] accumulation, GSH/GSSG depletion, MDA elevation, and GPX4 inactivation, in A549 and PC9 cells. NQO1 overexpression partially reversed these effects, whereas NQO1 knockdown phenocopied DHA-induced ferroptotic responses. GPX4 overexpression partially rescued the ferroptotic phenotype induced by NQO1 depletion, supporting a functional link between NQO1 and the GPX4 antioxidant axis. In vivo, DHA suppressed tumor growth, an effect partially reversed by NQO1 overexpression. DHA triggers ferroptosis in the NSCLC models examined, particularly in LUAD-derived cell models, in part through NQO1-mediated ROS burst and impairment of the GSH/GPX4 antioxidant defense. These findings identify NQO1 as a critical mediator of DHA-induced ferroptosis in these models and provide a mechanistic rationale for further investigation of DHA and related artemisinin derivatives in NSCLC.

4'-Hydroxywogonin, a bioactive flavonoid derived from Scutellaria baicalensis, has reported anti-inflammatory and anticancer activities; however, its therapeutic potential and molecular mechanism in hepatocellular carcinoma (HCC) remain unclear. This study investigated the anti-tumor effects of 4'-hydroxywogonin and its regulatory role in Gas6/Axl-associated signaling. Human HCC cells were used to evaluate proliferation, migration, invasion, cell cycle distribution, apoptosis, senescence, and epithelial-mesenchymal transition. Whole-transcriptome sequencing, western blotting, quantitative polymerase chain reaction, molecular docking, and thermal shift analysis were performed to explore the underlying mechanism, and antitumor efficacy and safety were further examined in a mouse xenograft model. 4'-Hydroxywogonin significantly inhibited HCC cell proliferation, migration, and invasion, while inducing G1-phase arrest, apoptosis, and senescence. Transcriptomic and molecular analyses identified Axl as a key affected target. 4'-Hydroxywogonin reduced Gas6 and Axl expression, suppressed PI3K/AKT activation, and downregulated cell cycle and epithelial-mesenchymal transition (EMT)-related proteins. Molecular docking predicted a potential interaction between 4'-hydroxywogonin and Axl, which was supported by thermal shift analysis. Axl overexpression partially rescued the inhibitory effects of 4'-hydroxywogonin, suggesting involvement of Axl-dependent and additional pathways. In vivo, 4'-hydroxywogonin markedly reduced tumor growth with efficacy comparable to sorafenib and without evident hepatotoxicity. These findings indicate that 4'-hydroxywogonin suppresses HCC progression, at least in part, through inhibition of the Gas6/Axl-PI3K/AKT signaling axis, supporting its further development as a natural therapeutic candidate.

Renal interstitial fibrosis (RIF) is a major driver of chronic kidney disease (CKD) progression, yet effective therapies remain limited. The Shen-Yan-Fang-Shuai formula (SYFSF) exhibits clinical renoprotective effects; however, its active components and mechanisms remain unclear. This study identifies notoginsenoside R2 (NGR2) as a core active compound of SYFSF that directly binds to caspase-2 (CASP2). Integrative multi-omics analyses in unilateral ureteral obstruction (UUO) rats and transforming growth factor-β1 (TGF-β1)-induced NRK-52E cells demonstrate that NGR2 upregulates CASP2, enhances the CASP2-aldehyde dehydrogenase 3 family member A1 (ALDH3A1) interaction, and promotes ALDH3A1 deubiquitination, thereby stabilizing ALDH3A1. This stabilization restores the integrity of mitochondria-associated endoplasmic reticulum membranes (MAMs) and improves fatty acid oxidation (FAO) by upregulating carnitine palmitoyltransferase 1A (CPT1A), acyl-CoA dehydrogenase long chain (ACADL), and peroxisome proliferator-activated receptor α (PPARα). It also reduces lipid accumulation, oxidative stress, epithelial-mesenchymal transition (EMT), inflammation, and collagen deposition. In vivo, NGR2 ameliorates renal dysfunction and fibrosis. Mechanistically, NGR2 binds the caspase activation and recruitment domain (CARD) of CASP2 without occluding the ubiquitin-interacting motif-like (UIML) region required for its deubiquitinase function, thereby increasing CASP2 abundance and stability rather than activating its apoptotic protease activity, and facilitating ubiquitin chain removal from ALDH3A1. These findings demonstrate that SYFSF and NGR2 antagonize RIF by targeting the CASP2/ALDH3A1 axis to restore MAM function and FAO, providing a molecular basis for traditional Chinese medicine in CKD therapy.

Salvia miltiorrhiza Bunge has been a commonly used medicinal herb in China since ancient times, widely employed to treat blood stasis-related disorders, including cardiovascular and inflammatory diseases. Dihydrotanshinone I (DHT), a lipophilic diterpenoid derived from its roots, has demonstrated anti-tumor potential; however, its specific molecular mechanisms involved in gastric cancer (GC) metastasis remain a mystery. Here, we report that DHT significantly suppresses the invasion and metastasis of GC by directly targeting macrophage migration inhibitory factor (MIF). Using limited proteolysis-mass spectrometry (LiP-MS), we identified MIF as a direct binding target of DHT, which was further validated by surface plasmon resonance and molecular docking. Mechanistically, DHT binding to MIF restored GSK-3β activity, promoted β-catenin phosphorylation (Ser33/37/Thr41), and facilitated its proteasomal degradation, thereby inhibiting β-catenin nuclear translocation and transcription of epithelial-mesenchymal transition-related genes. In vivo experiments confirmed that DHT markedly suppressed peritoneal metastasis of GC. These findings not only elucidate a novel molecular axis-DHT/MIF/GSK-3β/β-catenin-in suppressing GC metastasis but also provide a mechanistic basis for the ethnopharmacological application of Danshen in cancer-related disorders. Furthermore, our findings raise the hypothesis that MIF might serve as a potential predictive biomarker for DHT treatment response in GC, a concept that warrants future clinical validation.

Emerging as a significant clinical burden, metabolic dysfunction-associated steatohepatitis (MASH) continues to challenge global healthcare systems due to limited effective interventions. Magnoflorine, a naturally occurring quaternary aporphine alkaloid abundant in dietary botanicals, has demonstrated promising hypolipidemic and anti-inflammatory properties in preliminary investigations. This investigation systematically evaluated both the therapeutic potential and mechanistic pathways underlying the capacity of magnoflorine to ameliorate MASH-associated hepatic pathologies. Male C57BL/6 mice received 12-week high-fat and high-fructose diet (HFFD) induction to establish MASH models, with graded magnoflorine dosing. Through integrated histopathology (H&E/ORO/Masson), transcriptomics, and targeted interventions at both cellular and animal levels, we elucidated the therapeutic mechanisms of magnoflorine. Results showed that magnoflorine effectively alleviated hepatic steatosis, inflammation, and fibrosis in HFFD-fed mice. Mechanistically, RNA-seq revealed that the 5'-monophosphate AMP-activated protein kinase (AMPK) activation was the central pathway mediating the effects of magnoflorine to impede MASH progression. Furthermore, AMPKα activation induced by magnoflorine stimulated hepatic lipophagy in HFFD-fed mice, which further affected lipid β-oxidation and inflammation. The findings were consistent in HepG2 cells incubated with palmitic acid. In vitro, silencing AMPKα reversed the stimulatory effect of magnoflorine on lipophagy in HepG2 cells. In vivo, liver-specific AMPKα silencing in mice blocked the enhancement of hepatic lipophagy and the improvement of MASH caused by magnoflorine. In brief, this study indicates that magnoflorine improves MASH in mice by activating hepatic lipophagy via AMPKα-dependent effects. This discovery provides a novel potential therapeutic alternative for MASH.

N-acylethanolamines (NAEs) are endogenous fatty acid amides structurally related to the endocannabinoid anandamide and function as bioactive lipid mediators with broad regulatory roles in inflammation and neuroinflammation. Among pattern-recognition receptors, Toll-like receptor 4 (TLR4) occupies a central position in innate immune surveillance, sensing pathogen-associated molecular patterns, such as lipopolysaccharide (LPS), as well as damage-associated molecular patterns. Upon ligand engagement, TLR4 triggers both MyD88- and TRIF-dependent signaling cascades, leading to NF-κB activation and the transcriptional induction of pro-inflammatory cytokines and Type I interferons. Within this family, classical NAEs, such as palmitoylethanolamide (PEA), and structurally distinct NAE-like derivatives represent two partially overlapping but pharmacodynamically different groups of bioactive compounds. Classical NAEs, including palmitoylethanolamide (PEA), primarily exert their anti-inflammatory actions through activation of peroxisome proliferator-activated receptors and related nuclear pathways. In contrast, NAE-like compounds with distinct scaffolds, such as 2-pentadecyl-2-oxazoline (C15OXA) and N-palmitoyl-D-glucosamine (PGA), may interact with the TLR4/MD-2 complex and modulate receptor activation more proximally, as suggested by cell-based functional assays and computational modeling. PRISMA-guided analyses across experimental models of colitis, LPS-induced systemic inflammation, and alcohol binge exposure consistently demonstrate the anti-inflammatory efficacy of PGA and C15OXA, identifying TLR4 as a pivotal mechanistic node underlying their activity. Together, these findings delineate distinct yet partially convergent pharmacodynamic profiles among endogenous NAEs and naturally occurring NAE-like compounds, supporting their potential relevance as natural-product-inspired pharmacological strategies for the modulation of TLR4-associated inflammatory and neuroinflammatory processes.

Phloretin (PHL) is an effective monomer component of Phellodendri Chinensis Cortex, Spica Prunellae, and fleshy fruits and exerts therapeutic effects on rheumatoid arthritis (RA). However, the underlying mechanism is yet to be elucidated. Following PHL treatment, therapeutic benefits were evaluated in a collagen-induced arthritis (CIA) rat model using arthritic markers and joint pathology. Network pharmacology, transcriptomics, Western blotting, IHC, molecular docking, Biacore, and cellular thermal shift assay were employed. PHL administration considerably reduced joint swelling, synovial hyperplasia, and bone erosion. Integrated network pharmacology and transcriptomic analyses identified the hypoxia-inducible factor-1 alpha (HIF-1α) axis as a critical target. In vitro experiments indicated that PHL effectively inhibited TNF-α-induced fibroblast-like synoviocyte (FLS) proliferation, migration, invasion, and glycolytic reprogramming, promoted apoptosis, and suppressed excessive autophagy. Furthermore, in a coculture system, PHL substantially attenuated FLS-induced tube formation in human umbilical vein endothelial cells. Furthermore, PHL demonstrated high affinity for HIF-1α and accelerated the degradation of HIF-1α protein. Importantly, HIF-1α overexpression reversed PHL's inhibitory effect. RA severity was alleviated in Hif1a[+/-] mice, validating HIF-1α's pathogenic role in disease progression. PHL exerts anti-RA effects by targeting HIF-1α to modulate glycolysis and the autophagy-apoptosis balance in FLS, thereby inhibiting synovial angiogenesis. These findings provide a strong foundation for developing and applying PHL as a potential therapeutic agent for RA.

Butein (3,4,2',4'-tetrahydroxychalcone) is a bioactive chalcone with antioxidant, anti-inflammatory, and cytoprotective properties. However, its effects on pathological cardiac hypertrophy and the underlying molecular mechanisms remain unclear. In this study, neonatal rat ventricular cardiomyocytes stimulated with phenylephrine and mouse models of transverse aortic constriction (TAC) at 2 and 8 weeks were used to investigate the protective effects of butein in vitro and in vivo. Butein markedly reduced cardiomyocyte hypertrophy, decreased intracellular and mitochondrial reactive oxygen species, preserved mitochondrial membrane potential, and restored mitochondrial bioenergetic status. Mechanistically, butein enhanced nuclear accumulation of Nrf2, restored antioxidant gene expression, and reactivated AMPK-Sirt3-FoxO3a signaling while reversing SOD2 hyperacetylation. In vivo, butein suppressed cardiac hypertrophy during the early compensated stage (2week TAC) and alleviated cardiac remodeling, fibrosis, and functional deterioration during the advanced pathological stage (8-week TAC). Notably, genetic silencing of Nrf2 or Sirt3 and pharmacological inhibition of Nrf2 attenuated the protective effects of butein, supporting the involvement of Nrf2-Sirt3 signaling both in vitro and in vivo. Collectively, these findings demonstrate that butein mitigates pathological cardiac hypertrophy by restoring redox and mitochondrial homeostasis through coordinated Nrf2-Sirt3 signaling.

Diabetes-related depression (DD) is a challenging comorbidity characterized by self-reinforcing interactions between metabolic dysfunction and affective disturbance, while conventional antidepressants and glucose-lowering drugs often yield suboptimal outcomes with adverse effects. This study investigated the therapeutic potential of fingered citron extract (FCE), traditionally used for metabolic and inflammatory conditions, in alleviating DD and elucidated its underlying mechanisms. Using a DD mouse model induced by high-fat diet, streptozotocin, and chronic unpredictable mild stress, we administered FCE at varying doses and employed a multimodal approach including behavioral tests, biochemical assays, UPLC-MS/MS, network pharmacology, molecular docking, molecular dynamics simulation, immunofluorescence, Western blotting, and cellular validation. The results showed that FCE significantly reversed weight loss, reduced fasting blood glucose, and ameliorated depressive-like behaviors. It also restored 5-HT, DA, NE, and BDNF, lowered glutamate and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), and increased anti-inflammatory IL-10. UPLC-MS/MS analysis identified 34 serum-absorbed bioactive constituents, and integrated analyses highlighted neohesperidin (NH) and SRC as a candidate active constituent-target axis. FCE suppressed pancreatic SRC overactivation, improved pancreatic and brain insulin (INS) related signaling, and these molecular changes were associated with behavioral recovery. In vivo pharmacodynamic validation further showed that NH partially reproduced the metabolic, behavioral, and biochemical effects of FCE in DD mice. In high-glucose-induced MIN6 cells, NH enhanced insulin secretion, mitigated reactive oxygen species accumulation, and restored mitochondrial membrane potential, whereas these protective effects were markedly attenuated after SRC knockdown. Collectively, these findings suggest that FCE ameliorates DD, at least in part, through NH/SRC-mediated regulation of insulin signaling and neuroinflammation, supporting its potential as a multi-target natural therapeutic candidate for DD.

Ajugol (AJ), the primary component extracted from the root of Rehmannia glutinosa, exhibits protective effects against metabolic dysregulation and chronic inflammation. This present work aims to study the therapeutic effect of AJ on liver fibrosis and to explore the underlying mechanism. Liver fibrosis was induced in C57BL/6J mice through the administration of 15% CCl4, followed by intragastric dosing of AJ at 20 and 40 mg/kg. We conducted a comprehensive evaluation involving histopathological assessment, serum biochemistry, RNA sequencing, and targeted metabolomics. In vitro models consisted of DCA-stimulated AML12 and RAW264.7 cells. Compared to untreated fibrotic mice, AJ intervention decreased the levels of serum ALT and AST, as well as hepatic hydroxyproline, α-SMA, and COL1A1. Transcriptomic analysis revealed that AJ significantly regulated key genes involved in primary bile acid (BA) biosynthesis and arachidonic acid (ARA) metabolism. AJ downregulated CYP7A1 expression and reduced the accumulation of toxic deoxycholic acid (DCA) in the mouse liver. Additionally, AJ decreased the expressions of COX-2, PTGR1, PTGR2, and HPGDS, as well as the levels of downstream metabolites prostaglandins PGD2 and PGE2, thereby improving ARA metabolism via the COX pathway. The anti-inflammatory effects of AJ were evidenced by a decrease in pro-inflammatory cytokines IL-1β, IL-6, TNF-α, and macrophage markers such as F4/80 and CD11b. In vitro, AJ alleviated DCA-induced hepatocellular injury and inhibited pro-inflammatory cytokine production in macrophages. DCA inhibited nuclear FXR activation, impairing hepatocyte function and activating inflammatory response. AJ effectively restored these functions. AJ improved hepatic inflammation and fibrosis, partly by alleviating the harmful effects of toxic BA accumulation. This highlights AJ as a promising novel therapeutic agent for liver fibrosis.

Amomum villosum Lour. has been traditionally valued in various cultures for its health benefits. Here, we show that the neuroprotective effects of the essential oils extracted from the fruit of Amomum villosum Lour. (AVEO) and elucidate its underlying mechanisms in lipopolysaccharide (LPS)-induced neuroinflammation. The main compounds of AVEO can cross the blood-brain barrier and significantly improve cognitive and behavioral dysfunction in LPS-induced mice, leading to a 50% enhancement in Morris water maze performance, a 60% and 80% reduction in immobility time in the tail suspension test and forced swimming test, respectively. Moreover, AVEO mitigates neuron loss and suppresses the expression of pro-inflammatory mediators, including TNF-α, IL-1β, and IL-6, while upregulating the anti-inflammatory cytokine IL-10 in brain tissue. RNA-seq and metabonomics analyses reveal that AVEO modulates multiple inflammation-related pathways and restore metabolic homeostasis in brain tissue. Remarkably, AVEO attenuates hyperactivation of the PI3K-Akt and MAPK signaling pathways in LPS-induced microglia and improves neuronal damage in LPS-induced microglia-neuron co-culture system. Altogether, our findings suggest that AVEO alleviates neural dysfunction by modulating PI3K-Akt and MAPK signaling cascades and promoting metabolic reprogramming upon neuroinflammation, thereby revealing the therapeutic potential of AVEO in several neurological disorders.

Chronic kidney disease (CKD) is driven by persistent inflammation and progressive fibrosis, leading to irreversible renal damage. Gramine (GMN), an indole alkaloid, has broad pharmacological benefits. However, its therapeutic potential against renal fibrosis remains unexplored. This study investigates the antifibrotic potential of GMN in an experimental model of CKD. Potential therapeutic targets of GMN associated with renal fibrosis were identified through network analysis, and functional enrichment analysis revealed the mechanisms regulated by these targets. In vitro validation was conducted in transforming growth factor-β1 (TGF-β1)-induced normal rat kidney fibroblast (NRK-49F) cells. Fibrosis in the left kidney was induced by ureter obstruction, and mice received GMN orally at doses of 10 and 30 mg/kg. GMN targets 110 genes related to renal fibrosis that regulate profibrotic pathways, including TGF-β1-mediated canonical Smad signaling. GMN effectively attenuated TGF-β1-induced fibroblast-to-myofibroblast transition by inhibiting Smad signaling, resulting in downregulation of collagen I and fibronectin. In vivo, oral administration of GMN ameliorated the histopathological damage induced by UUO and significantly improved renal function. Mechanistically, GMN suppressed NF-κB-mediated priming of the NOD-like receptor pyrin domain-containing 3 (NLRP3) inflammasome and inhibited IL-1β maturation, thereby mitigating renal inflammation. UUO-induced fibrotic responses, characterized by upregulation of TGF-β1, TGF-β receptor type II, p-Smad2/3, connective tissue growth factor (CTGF), alpha-smooth muscle actin, and extracellular matrix (ECM) components were significantly attenuated by GMN, suggesting its ability to inhibit pathological ECM remodeling. This study demonstrates that GMN attenuates renal inflammation, suppresses myofibroblast activation and ECM remodeling by concurrently inhibiting NF-κB/NLRP3 inflammasome axis and TGF-β1/Smad pathway.