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.
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.
ABSTRACT 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.
ABSTRACT Diabetic osteoporosis (DOP) is characterized by chronic low‐grade inflammation and impaired bone formation. Pinoresinol diglucoside (PDG), a bioactive compound derived from Eucommia ulmoides Oliv., has shown therapeutic potential in metabolic bone disorders; however, its direct molecular targets and integrated mechanisms remain unclear. This study aimed to elucidate the metabolic basis and molecular targets through which PDG protects against high glucose (HG)‐induced osteoblast dysfunction and diabetic bone loss. Osteogenic effects of PDG were evaluated in HG‐exposed osteoblasts, with untargeted metabolomics, qPCR, Western blotting, molecular docking, SPR, and siRNA‐mediated knockdown applied to identify altered pathways and validate target necessity. In vivo, PDG was administered to db/db mice with DOP for 8 weeks, followed by micro‐CT, bone metabolism factors, histomorphometry, and immunofluorescence analyses. Metabolomic profiling identified arachidonic acid metabolism as the central pathway dysregulated in HG conditions and rescued by PDG. PDG directly bound and inhibited both p65 (NF‐κB) and GSK3β. Mechanistically, p65 knockdown abolished the anti‐inflammatory effect of PDG without affecting osteogenesis, whereas GSK3β knockdown abrogated β‐catenin activation and osteogenic differentiation without affecting anti‐inflammatory responses, indicating that they function as parallel and independent branches. Simultaneous knockdown of both completely blocked the functional recovery of osteoblast differentiation. In DOP mice, PDG significantly improved trabecular microarchitecture and bone metabolic homeostasis, reduced local PGE2 levels with an effect directly targeting bone, inhibited p65 nuclear translocation, and restored β‐catenin nuclear localization. PDG ameliorates DOP by dual targeting p65 and GSK3β, coordinately inhibiting NF‐κB‐mediated inflammation and activating the β‐catenin osteogenic program, highlighting its therapeutic potential for DOP.
