ABSTRACT 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.
ABSTRACT 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.
ABSTRACT 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.
ABSTRACT 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.
ABSTRACT 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% CCl 4 , 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.
