Search Results (2,016)

Immune stress under intensive high density production conditions impairs broiler health and performance, highlighting the need for effective nutritional strategies. This study established an immune stress model by intraperitoneal lipopolysaccharide (LPS) injection to investigate the mechanism by which Glycyrrhiza polysaccharides (GPs) alleviate LPS-induced immune stress. A total of 240 AA broilers were randomly assigned to four groups: control (CON), GP, LPS, and GP+LPS, with six replicates per group and ten birds per replicate. During the stress phase (28 d), LPS challenge increased serum ACTH and CORT concentrations and altered several immune-related parameters; whereas, GP supplementation improved ileal morphology and reduced serum IL-1β, NF-κB, and TNF-α concentrations. Significant GP × LPS interactions were observed for IL-4, IgA, IgG, and iNOS. During the recovery phase (35 d), GP supplementation increased serum IgA and decreased iNOS, while significant GP × LPS interactions were detected for IL-1β, NF-κB, and sCD4. Notably, GP attenuated the LPS-associated increase in NF-κB. Integrated multi-omics analysis indicated coordinated regulation at transcriptional and proteomic levels. GP upregulated FOS, JUN, and CATH3 and enriched NF-κB and Wnt signaling at the transcript level, while increasing IRF1 and CATH3 and decreasing CTNNBIP1 at the protein level, primarily involving Wnt and MAPK pathways. Joint analysis identified CATH3 as a central regulatory target. Collectively, GP alleviates immune stress by restraining excessive inflammation and modulating Wnt/MAPK signaling, thereby promoting intestinal structural recovery. Full article

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation and joint destruction. Cellular senescence contributes to chronic inflammation, yet key senescence-associated regulators in RA remain unclear. This study aimed to identify RA senescence-related signatures and their roles. Using GSE89408 as the training cohort, we screened hub genes by intersecting differentially expressed and senescence-related genes via WGCNA and three machine learning algorithms, with GSE55457 for external validation. Immune infiltration, regulatory network, subtyping and drug prediction were analyzed. Clinical and in vitro assays validated RIPK2 expression and function in the macrophage senescence-like phenotype, with preliminary signaling exploration. Three senescence-related hub genes (TNFAIP6, SLC2A3, RIPK2) were identified. The derived nomogram showed robust diagnostic performance (AUC = 0.988). Hub genes correlated strongly with myeloid cells, especially macrophages. Two immunologically distinct RA subtypes were identified. RIPK2 was upregulated in clinical samples; its inhibition attenuated LPS-induced macrophage senescence-like changes and inflammation. Preliminary data suggested RIPK2 may act via the NF-κB pathway. This study identifies RA senescence-associated signatures, revealing RIPK2 linking innate immunity to macrophage senescence-like changes, offering novel insights into pathogenesis and supporting it as a candidate biomarker and therapeutic target. Full article

Glutamine peptides not only serve as delivery vehicles for supplemental glutamine but also frequently possess unique biological activities that surpass glutamine itself, playing a crucial role in maintaining intestinal health. Corn gluten meal, as a major byproduct of corn processing with substantial production volume, is rich in glutamine, making it an ideal raw material for preparing glutamine peptides. This study aims to establish an enzymatic hydrolysis process for the efficient release of glutamine peptides from corn gluten meal and to identify glutamine peptides with gut health-maintaining effects. The results indicate that stepwise enzymatic hydrolysis of corn gluten meal using Protamex and Trypsin yields a glutamine-rich corn protein hydrolysate (GRCH). A total of 175 glutamine peptides were further identified from the low-molecular-weight fraction of GRCH. Through physicochemical property analysis and molecular docking technology, five glutamine peptides with potential inhibitory activity against the JAK2/STAT3 signaling pathway were screened, among which QFSLP demonstrated outstanding performance. In vitro experiments demonstrate that QFSLP is fully absorbed by intestinal epithelial cells and effectively alleviates LPS-induced inflammatory responses in intestinal cells. Following QFSLP intervention, the levels of proinflammatory factors TNF-α, IL-1β, and IL-8 in cells were significantly reduced (p < 0.05), while the level of the anti-inflammatory factor IL-10 was significantly increased (p < 0.05). The findings of this study contribute to the industrial-scale production of glutamine peptides and provide experimental evidence for their development in gut health-related products. Full article

Exosomes carry bioactive macromolecules driving sepsis pathogenesis, but the mechanisms underlying macrophage-to-hepatocyte communication during systemic inflammation remain poorly understood. We investigated how sepsis-induced macrophage exosomes are involved in remote intercellular crosstalk with hepatic cells via transcription factor-encoding mRNA cargo. Human monocytic THP-1 macrophages were stimulated with lipopolysaccharide (LPS), followed by exosome isolation, recipient cell uptake verification, and high-throughput RNA sequencing cargo analysis. To functionally reconstruct downstream signaling in recipient cells, exosome-enriched MAFB mRNA was transiently overexpressed in a HepG2 cell model, with subsequent expression changes mapped at both the transcript and protein levels using quantitative PCR and Western blot analyses. This ectopic MAFB expression directly upregulates the expression of microRNA-155 (miR-155). Crucially, elevated miR-155 acts as a post-transcriptional repressor that directly targets and downregulates JARID2 and H3F3A mRNAs and their corresponding protein products within the liver cells, orchestrating a “repressor-of-repressors” disinhibition cascade that drives net chromatin remodeling and activation of downstream hepatic target genes. This study demonstrates that sepsis alters exosomal transcription factor mRNA cargo and delineates a mechanistic downstream pathway—MAFB → ↑miR155 → ↓Jarid2 & ↓H3F3A → Chromatin Remodeling → Downstream Hepatic Gene Activation pathway—that provides novel, specific molecular checkpoints for therapeutic intervention in sepsis-induced liver injury. Full article

Osteoarthritis (OA) is a progressive joint disease characterized by cartilage degeneration, chronic low-grade inflammation, and disruption of tissue homeostasis. Although the mechanosensitive ion channel Piezo1 has been implicated in OA pathogenesis, its expression may also be modulated by inflammatory stimuli independently of applied mechanical loading. This study established a scaffold-free three-dimensional co-culture organoid model comprising human bone marrow-derived mesenchymal stem cell-derived chondrocyte-like cells and M-CSF/RANKL-differentiated RAW264.7-derived osteoclast-like cells to investigate Piezo1-associated molecular responses, inflammatory signaling, and mitophagy-related markers under lipopolysaccharide (LPS)-induced inflammatory conditions. Osteoclast-like differentiation was validated in parallel monolayer cultures by tartrate-resistant acid phosphatase staining and the presence of multinucleated cells before the corresponding differentiated cultures were used for organoid generation. Histological staining, immunofluorescence, CellTiter-Glo 3D viability assay, lactate dehydrogenase cytotoxicity assay, RT-qPCR, and Western blotting were used to evaluate extracellular matrix formation and inflammatory, catabolic, and mitochondrial quality-control-associated markers. LPS stimulation increased the expression of Piezo1, HIF-1α, phosphorylated CaMKII, NLRP3, cleaved Caspase-1, and MMP13, together with alterations in mitophagy- and autophagy-associated markers. Among the evaluated compounds, curcumin produced the greatest improvement in viability relative to the LPS-treated group and was selected for subsequent molecular analyses. Curcumin treatment was associated with reduced inflammatory and catabolic marker expression and partial preservation of cartilage-associated matrix markers. These findings demonstrate inflammation-associated changes in Piezo1 expression and related molecular markers but do not establish mechanically activated Piezo1 signaling or Piezo1-dependent causality. The organoid system therefore represents an exploratory LPS-induced inflammatory model exhibiting selected OA-relevant molecular and matrix-associated features. Full article

Despite the traditional application of Sophora moorcroftiana (Benth.) Baker seeds for liver disorders, the capacity of its 70% ethanol extract (SMS) to alleviate metabolic dysfunction-associated steatotic liver disease (MASLD) and the underlying gut–liver axis mechanisms remain unclear. In this study, a high-fat diet (HFD)-induced MASLD mouse model was established to investigate the protective effects of SMS and its regulatory role in the interplay among gut microbiota, short-chain fatty acids (SCFAs), and inflammation. Serum, intestinal, and hepatic samples were collected to evaluate inflammatory responses, intestinal barrier integrity, and hepatic lipid metabolism. Gut microbiota composition and SCFA profiles were analyzed using 16S rRNA sequencing and metabolomics. In LPS-stimulated Caco-2 cells, SMS reduced inflammatory cytokines and TLR4/MyD88/NF-κB-associated signaling. The results demonstrated that SMS markedly alleviated hepatic steatosis by reducing triglyceride synthesis and hepatocellular lipid accumulation. In addition, SMS promoted the proliferation of beneficial bacteria, including Bifidobacterium and Akkermansia, and increased the production of SCFAs, particularly butyrate. SMS also restored intestinal barrier integrity through upregulation of Occludin and Claudin-1, thereby reducing circulating lipopolysaccharide (LPS) levels. Furthermore, SMS attenuated inflammation by inhibiting activation of the TLR4/NF-κB signaling pathway. Collectively, these findings demonstrate that SMS alleviates MASLD through coordinated modulation of gut microbiota composition, SCFA metabolism, intestinal barrier function, and inflammatory responses, highlighting its potential as a therapeutic strategy targeting the gut–liver axis. Full article

Bacterial infection triggered excessive inflammatory responses, yet the mechanisms linking inflammatory activation to immunometabolic adaptation remained incompletely understood. The mitochondrial translocator protein (TSPO) has been implicated in inflammatory activation and cellular metabolism. This study aimed to investigate the role of TSPO in inflammation mediated by Toll-like receptor 4 (TLR4). Herein, we integrated transcriptomic data from the human peripheral blood dataset GSE72829, and single-cell transcriptomic profiles from the CELLxGENE platform with cellular mechanistic experiments in BV2 microglia and RAW264.7 macrophages. Transcriptomic analyses revealed that TSPO expression was markedly upregulated in patients with bacterial infection (n = 52) and exhibited diagnostic potential to distinguish bacterial infection from healthy controls (HCs, n = 16) and viral infection (n = 92). TSPO-correlated genes were enriched in Toll-like receptor (TLR) signaling, inflammatory response, and immunometabolic pathways. Mechanistically, TSPO interacted with TLR4 and selectively modulated TLR4-driven inflammatory activation. TSPO deficiency augmented lipopolysaccharide (LPS) induced tumor necrosis factor‑α (TNF-α) and interleukin‑6 (IL-6) secretion, accompanied by disrupted Ca²⁺ homeostasis, impaired cholesterol balance, and compensatory metabolic remodeling characterized by elevated L-lactate and sustained Adenosine triphosphate (ATP) levels. Collectively, these findings identified TSPO as an immunometabolic regulator bridging TLR4 signaling and metabolic adaptation during inflammatory activation. Besides, TSPO represented a promising biomarker and therapeutic target to limit exaggerated inflammatory responses. Full article

Background: Lipid mediators (LM), comprising 17S-hydroxy-docosahexaenoic acid, resolvin D5, and protectin DX in a ratio of 3:47:50, were naturally generated from docosahexaenoic acid by soybean lipoxygenase and exhibit potent anti-inflammatory activities. However, their therapeutic potential in periodontitis (PD) remains unclear. Therefore, this study investigated the protective effects of LM against PD in vitro and in vivo. Methods: Human gingival fibroblasts (HGFs) were stimulated with Porphyromonas gingivalis lipopolysaccharide (PG-LPS) to evaluate the anti-inflammatory effects of LM. Nitric oxide (NO) production, prostaglandin E2 (PGE2) level, inflammatory cytokines secretion, matrix metalloproteinase 9 (MMP9) expression, and NF-κB/STAT3 signaling pathways were analyzed. In vivo, a ligature-induced rat model of PD was established, and LM were orally administered at doses of 5, 10, and 20 μg/kg once daily. The effects of LM were evaluated by histopathological examination, micro-computed tomography (micro-CT), quantitative real-time PCR (qRT-PCR), and immunohistochemistry (IHC). Results: In PG-LPS-stimulated HGFs, LM significantly reduced NO and PGE2 production by downregulating inducible nitric oxide synthase and cyclooxygenase-2 expression. Moreover, LM decreased the levels of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), IL-1β, and MMP9, accompanied by suppression of the NF-κB and STAT3 signaling pathways. In the ligature-induced rat model of PD, LM significantly attenuated gingival inflammation and alveolar bone loss. Furthermore, IHC analysis revealed that LM markedly reduced the level of IL-6, TNF-α, MMP9, and phosphorylated STAT3 in gingival tissues. Conclusions: LM exerted protective effects against periodontitis by alleviating inflammatory response and reducing alveolar bone loss, which may be associated with suppression of the NF-κB and STAT3 signaling pathways. Full article

Neutrophils are key effectors of innate immunity. Although traditionally regarded as short-lived effector cells, recent evidence suggests they can undergo functional reprogramming after microbial stimulation, leading to trained immunity. We investigated whether lipopolysaccharide (LPS) or Candida albicans β-glucan induces memory-like inflammatory responses in adult peripheral and neonatal cord blood neutrophils. We used complementary molecular and functional approaches to characterize neutrophil responses following in vitro priming with LPS or β-glucan and subsequent LPS restimulation. Cytokine secretion, reactive oxygen species (ROS) production, glycolytic activity (via lactate production and glycolytic enzyme expression), and ERK1/2 and NF-κB signaling pathway activation were assessed. Priming with LPS or β-glucan enhanced neutrophil responsiveness to secondary stimulation, resulting in increased secretion of IL-1β, IL-6, IL-8, CXCL1, and CCL2, together with elevated ROS production. These effects were consistently more pronounced in neonatal than in adult neutrophils. Memory-like neutrophils also exhibited metabolic reprogramming, associated by increased hexokinase-2 and phosphofructokinase-1 mRNA expression, elevated lactate production, and enhanced ERK1/2 and NF-κB activation. Human neutrophils, particularly neonatal cells, develop memory-like characteristics following LPS or β-glucan priming, highlighting their functional and metabolic plasticity and suggesting a role for neutrophil training in early-life immune adaptation. Full article

Background and Objectives: Oxidized LDL (oxLDL) and Toll-like receptor (TLR) signaling are implicated in inflammatory bone loss and periodontitis, but their cooperative effects on osteoclastogenesis remain unclear. We investigated whether TLR2/4 stimulation regulates the transcription of LOX-1 (lectin-like oxidized low-density lipoprotein receptor-1), a receptor for oxLDL, and whether oxLDL enhances TLR-mediated osteoclastogenic responses in the absence of exogenous RANKL. Materials and Methods: Mouse bone marrow cells (BMCs) were differentiated into bone marrow macrophages (BMMs) with M-CSF and stimulated with TLR ligands (Pam3CSK4 for TLR2 and Lipid A for TLR4) and/or oxLDL, with or without exogenous RANKL. Raw264.7 cells were stimulated with TLR ligands or Porphyromonas gingivalis lipopolysaccharide (LPS). LOX-1 transcriptional activity and osteoclastogenic responses were evaluated using luciferase assays, quantitative RT-PCR, and tartrate-resistant acid phosphatase (TRAP) staining. Results: In Raw264.7 cells, stimulation with Pam3CSK4, Lipid A, or LPS significantly increased LOX-1 promoter activity. In BMCs, Pam3CSK4 significantly increased LOX-1 and MyD88 mRNA expression on day 1, whereas expression decreased by day 3; these changes were suppressed by a TLR2 inhibitor. Lipid A significantly increased LOX-1 and MyD88 mRNA expression on day 3, and the increase in LOX-1 expression was inhibited by a TLR4 inhibitor. Functionally, Pam3CSK4 or Lipid A alone increased mononuclear TRAP-positive cells but did not induce multinucleated TRAP-positive cells. In contrast, co-stimulation of oxLDL with either Pam3CSK4 or Lipid A promoted multinucleated TRAP-positive cell formation even in the absence of exogenous RANKL. Conclusions: TLR2/4 stimulation transiently enhanced LOX-1 transcription and expression in association with increased MyD88 expression. TLR ligands cooperated with oxLDL to promote multinucleated TRAP-positive cell formation in the absence of exogenous RANKL, suggesting that inflammatory and metabolic signals may act synergistically during osteoclastogenesis. These findings improve our understanding of how dyslipidemia-associated factors may contribute to inflammatory bone-resorptive diseases, including periodontitis. Full article

Accurate estimation of terrestrial water storage change (TWSC) remains challenging in regions where hydrological variability interacts with complex geological conditions and intensive human activities. Taking the Loess Plateau (LP) in the middle Yellow River region as a case study, this work integrates GLDAS simulations, GRACE observations, GNSS vertical-displacement records, a joint GNSS–GRACE inversion, and meteorological data for 2013–2024 to investigate regional TWS variability and model-dependent discrepancies. The results show that GLDAS, GRACE, GNSS, and the joint solution exhibit distinct temporal trends and spatial patterns. GRACE indicates a stronger long-term depletion signal, whereas GNSS-derived equivalent water height (EWH), which relies on the assumption of elastic surface loading, shows a weaker trend but stronger seasonal variability. This discrepancy suggests that GNSS-based inversion over the LP may be affected by non-elastic or non-loading deformation processes, such as wetting-induced loess collapse, aquifer compaction, mining-related subsidence, and other near-surface effects. In contrast, GRACE may include non-TWS mass redistribution associated with soil erosion and mineral exploitation. The joint solution is more consistent with the GLDAS-derived hydrological model benchmark than either single geodetic estimate, but this agreement should not be interpreted as direct proof of higher accuracy or complete removal of non-hydrological effects. Overall, this study highlights the need to diagnose model-dependent discrepancies, effective spatial resolution, and non-loading deformation when applying GRACE- and GNSS-based approaches to TWSC estimation in geologically and anthropogenically complex regions. Full article

Background: Atopic dermatitis (AD) is a persistent and recurring skin disease characterized by epidermal barrier dysfunction, immune dysregulation, and elevated expression of proinflammatory mediators. We investigated the anti-inflammatory potential of polynucleotides (PN), highly purified DNA biopolymers isolated from salmonid gonads, in keratinocyte and macrophage activation models. Methods: RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS), whereas HaCaT keratinocytes were stimulated with tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). The effects of PN treatment on the production or expression of inflammatory mediators, cytokines, and chemokines were evaluated. Changes in the phosphorylation of mitogen-activated protein kinases (MAPKs) and Janus kinase 1/signal transducer and activator of transcription 3 (JAK1/STAT3) and in the nuclear localization of nuclear factor-κB (NF-κB) were also assessed. Results: In LPS-activated RAW 264.7 macrophages, PN treatment significantly suppressed nitric oxide production and downregulated the expression of inducible nitric oxide synthase (iNOS), TNF-α, IL-1β, and IL-8, accompanied by reduced NF-κB nuclear translocation. In TNF-α/IFN-γ-stimulated HaCaT keratinocytes, PN treatment markedly decreased the secretion levels of IL-6, IL-1β, and thymic stromal lymphopoietin. Moreover, PN treatment markedly reduced T-cell-recruiting chemokines, including MDC/CCL22, TARC/CCL17, RANTES/CCL5, and IL-8. Signaling analyses demonstrated that PN treatment attenuated the phosphorylation of key MAPKs (ERK, JNK, and p38) and the JAK1/STAT3 axis. Furthermore, PN treatment markedly reduced NF-κB nuclear translocation. Conclusions: These in vitro findings indicate that the anti-inflammatory effects of PN are associated with reduced activation of multiple core signaling pathways governing cytokine and chemokine responses, supporting further investigation of PN in AD and other chronic inflammatory skin diseases. Full article

Jing Guan Fang (JGF) is a traditional multi-herb formula used for hyperinflammatory conditions associated with severe viral infections; however, its protein-level effects in macrophage-mediated inflammation remain incompletely characterised. In this study, we combined ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS)-based phytochemical profiling, functional anti-inflammatory assays, and discovery-level proteomic and secretome analyses to investigate the anti-inflammatory effects of JGF in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Phytochemical profiling revealed predominantly flavonoid- and phenolic-related annotated features, together with selected terpenoid- and alkaloid-related annotations. Functionally, JGF treatment was associated with reduced nitric oxide production and differential effects on pro-inflammatory cytokine release, with a clearer concentration-dependent reduction in interleukin-6 (IL-6) and a more variable tumour necrosis factor-α (TNF-α) response, without reducing cell viability. Label-free quantitative proteomics suggested that JGF treatment was associated with modulation of LPS-induced protein changes, including reduced abundance of inflammation-associated proteins such as myristoylated alanine-rich C-kinase substrate (MARCKS) and inducible nitric oxide synthase (NOS2), together with partial restoration of selected regulatory proteins such as AIMP1 and AIMP2. Secretome analysis further suggested that JGF reduced extracellular inflammatory and tissue-remodelling-related mediators, including SERPINE1/PAI-1. Exploratory pathway analysis indicated that JGF treatment was associated with changes in inflammation-related and oxidative stress-related signalling networks. Collectively, these findings provide discovery-level molecular insights into the anti-inflammatory effects of JGF in LPS-stimulated macrophages and support further targeted validation of its pharmacological activity as a candidate multi-component anti-inflammatory formula. Full article

The BMP2-Smad signaling pathway serves as a central regulator of osteoblast differentiation and bone formation, rendering it a promising target for the discovery of osteogenic agents from natural sources. Nonetheless, direct BMP2-binding ligands derived from complex herbal extracts remain poorly characterized. In the present study, surface plasmon resonance (SPR)-based target fishing against BMP2, in conjunction with UPLC-Q-TOF-MS identification, was employed to screen for bioactive ligands from Periploca forrestii, a traditional Miao medicinal plant used for bone-related conditions. Six caffeoylquinic acid derivatives, namely neochlorogenic acid (NCA), 3-O-caffeoyl-4-O-sinapoylquinic acid, chlorogenic acid (CA), cryptochlorogenic acid (CCA), isochlorogenic acid B (IB), and isochlorogenic acid C (IC), were captured as direct BMP2-binding ligands. All six compounds promoted osteoblast differentiation and mineralization in MC3T3-E1 Subclone 14 cells, with IB displaying the strongest binding affinity and bioactivity. Mechanistically, IB failed to rescue the osteogenic suppression induced by the BMP type I receptor inhibitor LDN-193189, indicating its dependence on BMP signaling. In an LPS-induced inflammatory model, IB significantly reversed the downregulation of key proteins in the BMP2-Smad pathway (p-Smad1, Smad4, and Runx2) and osteogenic marker genes (Osterix, COL1A1, and OCN), demonstrating its capacity to restore osteogenic function under compromised conditions. Collectively, these findings establish that caffeoylquinic acid derivatives, particularly IB, function as BMP2-targeting osteogenic constituents of P. forrestii that activate BMP2-Smad signaling to promote osteoblast differentiation, thereby offering a pharmacological basis for the development of natural product-derived osteogenic agents. Full article

Background/Objectives: Heracleum millefolium Diels (HMD) is a traditional Tibetan medicinal herb that has been reported to possess analgesic, anti-edematous, and anti-inflammatory activities. However, its chemical composition and anti-inflammatory mechanisms remain unclear. Methods: In this study, ultra-performance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-HRMS), combined with database searching and “seed”-based molecular networking, was used to systematically characterize the chemical constituents of HMD. Furthermore, an integrated strategy combining network pharmacology, molecular docking, and cellular validation was applied to explore potential anti-inflammatory mechanisms associated with HMD. Results: A total of 472 compounds were identified or tentatively identified from HMD. Network pharmacology analysis predicted five key targets, namely TNF, IL-6, IL-1β, GAPDH, and AKT1, together with four potential bioactive constituents, including velutin, artemitin, kaempferol, and naringenin. Pathway enrichment analysis indicated that the potential anti-inflammatory effects of HMD were mainly associated with lipid and atherosclerosis and the AGE-RAGE signaling pathway. Molecular docking suggested potential favorable interactions between the selected constituents and key targets. Moreover, cellular experiments demonstrated that the four compounds significantly inhibited inflammatory responses in LPS-stimulated RAW264.7 macrophages. Conclusions: This study systematically characterized the chemical profile of HMD and preliminarily explored its potential anti-inflammatory mechanisms, providing a scientific basis for further investigation of its bioactive constituents and pharmacological properties. Full article