Search Results (920)

Osteoarthritis (OA) is a chronic degenerative joint disease characterized by persistent low-grade inflammation and progressive cartilage destruction. Macrophage-driven inflammatory responses contribute to extracellular matrix (ECM) degradation and accelerate disease progression. Here, we investigated the therapeutic potential of a DHA-derived lipid mediator mixture (LM), generated via soybean lipoxygenase and composed of 17S-hydroxydocosahexaenoic acid, resolvin D5, and protectin DX (3:47:50), in regulating macrophage–chondrocyte crosstalk and OA progression. LM significantly reduced IL-6, IL-1β, and TNF-α production in lipopolysaccharide-induced THP-1 macrophages. Conditioned medium from LM-treated macrophages attenuated ECM degradation in primary chondrocytes by suppressing MMP13 and ADAMTS5 while restoring COL2A1 and ACAN expression, indicating that LM may indirectly protects ECM by modulating the inflammatory microenvironment. In parallel, LM directly protected chondrocytes against IL-1β-induced inflammatory and catabolic responses, and restored ECM homeostasis. Mechanistically, LM significantly increased SIRT1 expression and deacetylation activity, as demonstrated by reduced NF-κB p65 acetylation. Both pharmacological inhibition by EX527 and siRNA-mediated SIRT1 knockdown abolished the protective effects of LM on ECM preservation. In vivo, LM oral administration alleviated cartilage destruction, improved joint structure and suppressed OA progression in a monosodium iodoacetate-induced OA model. Notably, micro-CT studies have demonstrated that LM significantly improved subchondral bone architecture, as evidenced by increased bone volume fraction and improved trabecular parameters. Histological analyses confirmed that LM attenuated inflammation and maintained cartilage integrity. Consistently, immunohistochemical findings showed reduced MMP13 expression, restoration of collagen II and aggrecan, and upregulation of SIRT1 in the LM-treated group compared to OA rats. Collectively, these findings suggest that LM mitigates OA progression by reducing inflammation, preserving ECM homeostasis, and attenuating subchondral bone deterioration. Full article

Venous thromboembolism (VTE) is a major cause of morbidity and mortality, underscoring the need for new molecular markers to enable early detection and clarify underlying mechanisms. Iron metabolism is linked to oxidative stress, endothelial injury, and inflammation, all central to thrombosis, yet its transcriptomic contribution to VTE remains unclear. We analyzed gene expression profiles from GSE19151 and GSE48000 using differential expression and weighted gene co-expression network analysis (WGCNA), integrating results with an iron metabolism gene set. Three hub genes were identified, arachidonate 5-lipoxygenase (ALOX5), Rho GTPase activating protein 1 (ARHGAP1), and glucose-6-phosphate dehydrogenase (G6PD), all downregulated in VTE. Gene set enrichment indicated that ALOX5 is involved in endothelial regulation, lipid metabolism, and immune pathways. A three-gene signature showed high diagnostic accuracy (AUC = 0.924 in the discovery cohort; 0.705 in validation). Immune deconvolution revealed broad immune remodeling and associated ALOX5 with multiple immune cell subsets, especially M0 macrophages, and with regulators such as TGFB1 and IL6R. Western blot analysis further showed that ALOX5 protein expression was significantly increased in LPS-activated HUVECs, supporting its involvement in inflammatory endothelial injury. DrugBank screening identified 19 approved drugs targeting ALOX5, supporting its potential for mechanistic and clinical investigation. Full article

The Senegal tree (Sengalia senegal) is the primary plant source of Gum Arabic (GA), a natural secretion rich in soluble fiber and bioactive polysaccharides. It has longstanding uses in traditional medicine, nutrition, and pharmaceuticals. The present study aimed to evaluate the phytochemical profile, antimicrobial, anti-inflammatory, and anticancer activities of GA methanolic extract (GAME), supported by molecular docking analysis of its key compounds. The gas chromatography–mass spectrometry (GCMS) analysis of the GAME identified many compounds, such as 9-octadecenoic acid (38.29%), methyl ester (15.52), 1,2-benzenedicarboxylic acid, 3-nitro (9.8%), hexadecadienoic acid, methyl ester (8.5), and á-d-mannofuranoside, methyl (7.38). The molecular docking analysis showed that 9-octadecenoic acid had strong binding affinity with target proteins, which included xanthine oxidase (XO), lipoxygenase (LOX), and cyclooxygenase-2 (COX-2), with the highest affinity to XO (−137.03 kcal/mol) and lipoxygenase (−135.09 kcal/mol). GAME possessed broad-spectrum antibacterial activity against Salmonella typhimurium (S. typhimurium), Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), and Staphylococcus aureus (S. aureus), with a zone of inhibition from 16.28 to 16.93 mm. B. subtilis was resistant to the tested extract. The extract also showed good membrane stability and potent inhibition of albumin, XO, LOX, and COX-2, with IC₅₀ values of 31.62, 13.02, 27.6, and 28.99 μg/mL, respectively. The cytotoxic assessment demonstrated moderate, dose-dependent effects on the Caco-2 (colorectal adenocarcinoma) and HeLa (cervical carcinoma) cell lines. These findings highlight the therapeutic potential of GA as a natural plant source of antibacterial, anti-inflammatory and anticancer agents. The combination of molecular docking with in vitro assays provides strong evidence supporting its application in the development of plant-based pharmaceuticals. This research suggests that GA could be a useful ingredient in the creation of anti-inflammatory and antibacterial drugs derived from plants. Full article

Background/Objectives. The modern Western diet (MWD) provides high linoleic acid (LA) exposure, typically contributing 6–9% of the total caloric intake. These high LA levels have fueled a longstanding debate about whether this dietary pattern confers benefit or risk. Importantly, LA intake is disproportionately elevated among lower socioeconomic populations due to greater reliance on industrial seed oils and ultra-processed foods. Despite decades of research, controlled dietary intervention studies directly evaluating the biological consequences of varying LA exposure remain limited. Methods. The current randomized, double-blind intervention (ClinicalTrials.gov; NCT02962128; 11 November 2016) compared the effects of a 12-week Low-LA diet (2.5% energy) versus a High-LA diet (10.0% energy) in healthy adults. Outcomes included plasma concentrations of highly unsaturated fatty acids (HUFAs) and ex vivo zymosan-stimulated whole-blood oxylipin generation. Results. Fifty-two participants completed the intervention. High LA exposure resulted in marked reductions in plasma n-3 eicosapentaenoic acid (EPA) and eicosatetraenoic acid (ETA) concentrations compared with the Low-LA arm. Docosapentaenoic acid (DPA) was also significantly lower in weeks 4 and 8. In contrast, levels of the n-6 HUFA arachidonic acid (ARA) did not differ with dietary LA exposure. Conclusions. HUFA and oxylipin analyses revealed that higher dietary LA markedly increased the ratios of ARA to EPA and ARA- to EPA-derived oxylipin species, shifting the lipid mediator balance toward a more n-6-dominant inflammatory profile. Full article

Background: Root rot caused by Fusarium solani is a devastating disease in Angelica sinensis (danggui), leading to severe yield and quality losses. Sustainable control strategies are urgently needed. According to the plant “cry for help” theory, plants under pathogen attack may recruit beneficial microbes via root exudates. However, whether A. sinensis employs this strategy against F. solani remains unknown. This study aimed to identify potential “cry for help” metabolites and evaluate their biocontrol potential. Methods: LC-MS analysis revealed that F. solani infection significantly altered the metabolic profiles of both A. sinensis roots and rhizosphere soil. Results: Comparative analysis identified seven metabolites specifically upregulated in infected plants but not detected in the pathogen, including taurine, oxoadipic acid, quinolinic acid, 6-phosphogluconic acid, methyl cinnamate, 2-phenylethanol, and (R)-3-hydroxybutyric acid. Exogenous application of these seven metabolites revealed that taurine and methyl cinnamate significantly alleviated disease symptoms, improved plant growth (root length, biomass), and enhanced the activities of key defense enzymes (peroxidase, POD, phenylalanine ammonia-lyase, PAL, lipoxygenase, LOX, polyphenol oxidase, PPO). Furthermore, taurine and methyl cinnamate reshaped the rhizosphere microbiome. The incidence of root rot was reduced by 51.3% and 50.8%, respectively. Taurine enriched actinobacteria (e.g., Paeniglutamicibacter) and reduced the relative abundance of pathogenic Ascomycota fungi, while methyl cinnamate markedly enriched the nitrogen-fixing bacterium Azotobacter and the saprophytic fungus Schizothecium. Crucially, both treatments significantly suppressed the proliferation of F. solani in the rhizosphere. Conclusions: Our findings demonstrate for the first time that A. sinensis activates a “cry for help” response upon attack by F. solani, with taurine and methyl cinnamate preliminarily identified as key signaling metabolites that can directly or indirectly inhibit the development of A. sinensis root rot. These compounds enhance plant resistance and recruit beneficial microorganisms, offering a novel and promising ecological strategy for the green control of A. sinensis root rot. Full article

Oxidative stress and inflammation play a key role in many diseases. This study evaluated the potential of bioactive compounds from Red-belted Bracket and Artist’s Bracket mushrooms to mitigate these processes. Multistep extraction yielded fractions with diversified composition (triterpenoids, polysaccharides) and bioactivities, including antioxidant properties and inhibition of pro-inflammatory enzymes. Both species were rich in triterpenoids: ethanolic extracts from Artist’s Bracket contained mainly ganoderenic and ganoderic acids (≈31 μg/g d.w.), while Red-belted Bracket extracts contained phenolic acids (≈20 μg/g d.w., mainly vanillic and chebulic acids) and triterpenoids (≈73 μg/g d.w., mainly forpinic and formipinic acids). The alkaline and ethanolic extracts exhibited the highest radical scavenging and reducing activities. Lipoxygenase was inhibited only by ethanolic extracts, with IC₅₀ values of 0.93 mg d.w./mL for Artist’s Bracket (mixed inhibition) and 0.62 mg d.w./mL for Red-belted Bracket (noncompetitive). Artist’s Bracket was also a potent source of xanthine oxidase inhibitors acting uncompetitively (IC₅₀ = 0.71, 1.39, and 2.06 mg d.w./mL for ethanolic, methanolic, and aqueous extracts, respectively). In contrast, Red-belted Bracket was less active (IC₅₀ = 3.84 mg d.w./mL, noncompetitive). In conclusion, these mushrooms, particularly their ethanolic extracts, are promising sources of compounds with antioxidant and anti-inflammatory activities, acting as effective inhibitors of lipoxygenase and xanthine oxidase. Full article

Heat stress (HS), increasingly intensified by climate change, severely restricts tomato growth and productivity. Although core heat shock factor-mediated transcriptional networks have been extensively characterized, how lipid metabolic reprogramming is transcriptionally coordinated during thermotolerance remains unclear. Using SlHDZ19 overexpression and mutant lines together with transcriptomic, biochemical, promoter-binding, and gene-silencing analyses, we show that the homeobox-leucine zipper transcription factor SlHDZ19 promotes tomato (Solanum lycopersicum) thermotolerance by activating a PLA2-dependent lipid-metabolic transcriptional program in leaves. SlHDZ19 overexpression generally improved heat-stress performance, while SlHDZ19 mutant lines exhibited heightened sensitivity, including more severe wilting, higher electrolyte leakage, and reduced proline accumulation and CAT activity under heat stress. Transcriptomic analysis revealed that SlHDZ19 is required for the full induction of canonical heat-responsive genes and that the linoleic acid metabolism pathway was repeatedly implicated in SlHDZ19-dependent transcriptional changes. SlHDZ19 binds to and activates the promoters of SlPLA_2α, which encodes a phospholipase A2 involved in releasing linoleic acid from membrane lipids, and three lipoxygenase genes (SlLox7, SlLox8, and SlLoxC), accompanied by elevated overall PLA2 and LOX activities in SlHDZ19-overexpressing plants. Moreover, genetic silencing of SlPLA_2α in both wild-type and SlHDZ19-overexpressing backgrounds supported its functional requirement downstream of SlHDZ19 in thermotolerance. Collectively, our findings support a thermotolerance module in which SlHDZ19 transcriptionally regulates PLA2- and LOX-associated steps of linoleic acid metabolism, potentially linking lipid-associated signaling and membrane remodeling with heat stress adaptation in tomato. Full article

Black mustard (Brassica nigra L.) seeds are a rich source of bioactive phytochemicals; however, their antioxidant, antibacterial, and anti-inflammatory potential has not been comprehensively explored. Therefore, this study aimed to assess antioxidant, antibacterial, and anti-inflammatory effects of black mustard seed extracts obtained from Soxhlet extraction with hexane (HE) and ethanol (EE), and ultrasonic-assisted ethanolic (UE) extraction. HPLC analysis confirmed the presence of sinapic acid in all extracts. Phytochemical profiling revealed that the EE was enriched in phenolic compounds, while the UE exhibited a higher flavonoid content. Accordingly, both EE and UE demonstrated strong antioxidant activities, including radical scavenging capacity, reducing power, and inhibition of lipid peroxidation. All extracts demonstrated antibacterial activity against Staphylococcus epidermidis. The anti-inflammatory potential of extracts was supported by the inhibition of lipoxygenase and protease. The UE showed the strongest lipoxygenase inhibition, while the EE and UE exhibited comparable protease inhibitory effect. Regarding RAW264.7 cells, the extracts were non-cytotoxic and reduced the expression of IL-6 and IL-31. Molecular docking analysis suggested that sinapic acid contributes to the anti-inflammatory activity through interactions with key inflammatory targets. Overall, the EE and UE demonstrated multitarget antioxidant, antibacterial, and anti-inflammatory activities, supporting their potential application in functional and dermatological formulations for inflammation management. Full article

A series of coumarin–amino acid hybrids of glycine, γ-aminobutyric acid, and L-glutamic acid was developed. These compounds were evaluated for their antioxidant and anti-inflammatory activities in vitro and for their drug-likeness in silico. Antioxidant activity was assessed in vitro using the AAPH-induced linoleic acid peroxidation assay. Soybean lipoxygenase and ovine cyclooxygenase 2 were used in vitro to test the inhibitory activity of the adducts. An in silico evaluation was performed using the open-access platforms Molinspiration, SwissADME, PreADMET, Molsoft, GLORYx, CypRules, and LiverTox Workspace. The synthesis of the compounds proceeded via a facile procedure through the corresponding acid in very good yields. The antioxidant activity of the compounds is shown to be highly dependent on the linkage used, with compound 15 presenting the highest activity (93% inhibition). The most active LOX inhibitor is compound 4 (IC₅₀ = 58 μM), while compounds 4 and 5 are the most potent COX-2 inhibitors (IC₅₀ = 55 μM for both). Compounds 4, 9, and 15 are depicted as pleiotropic molecules (compound 4: IC₅₀ for SLOX-1 = 58 μM and IC₅₀ for COX-2 = 55 μM; compound 9: IC₅₀ for COX-2 = 60 μM, and 59% antilipid peroxidation; compound 15: IC₅₀ for COX-2 = 70.5 μM, 93% antilipid peroxidation). An in silico evaluation showed favorable properties of the designed agents, which were quantified, with all the compounds showing a QED score higher than 0.5. The overall results highlight that compound 4 can be used as a lead molecule for the design of more potent agents with a pleiotropic profile. Full article

Arachidonate 5-lipoxygenase (ALOX5) is a key enzyme implicated in several inflammatory disorders, including asthma and allergic rhinitis. Despite its therapeutic importance, only one compound is currently approved as an ALOX5 inhibitor in the United States, highlighting the urgent need for new drug candidates. Progress in this area is often hindered by conventional bioassays, which can be labor-intensive, costly, and unsuitable for complex mixtures. To overcome these challenges, we developed a simple thin-layer chromatography (TLC) bioautographic assay for the rapid detection of ALOX5 inhibitors in natural extracts, a rich source of pharmacologically active compounds. The method exploits the oxidative coupling of 3-methyl-2-benzothiazolinone hydrazone (MBTH) with 3-(dimethylamino)benzoic acid (DMAB) during the ALOX5-catalyzed conversion of arachidonic acid, producing a colored indamine dye. Experimental parameters influencing chromogenic reaction were investigated and optimized to minimize reagent consumption while ensuring accuracy and sensitivity of the method. The assay was then applied to a panel of natural products and to crude mushroom extracts, enabling the rapid identification of several active compounds within complex extracts, including the dual COX2/ALOX5 inhibitor 3α-acetylpolyporenic acid A. Easy to implement, cost-efficient, and well suited for screening and bioguided fractionation, this TLC bioassay provides a powerful tool to accelerate the discovery of novel anti-inflammatory compounds. Full article

Artichoke by-products (ABP) represent valuable sources of bioactive compounds with relevant health benefits. In this study, a green extraction strategy based on pressurized liquid extraction (PLE) was optimized to enhance the recovery of phenolic and flavonoid compounds from ABP using a response surface methodology. Extraction temperature and solvent composition were identified as the key factors driving extraction performance. Optimal conditions using a mixture of ethyl acetate and ethanol (90/10, v/v) at 180 °C significantly enhanced extraction yield, total phenolic and flavonoid content, and antioxidant activities, as measured by ORAC and DPPH assays. Chemical characterization via HPLC-C18-Q-TOF-MS/MS revealed a diverse profile of phenolic and flavonoid compounds, including caffeoylquinic acid derivatives and related transformation products. The neuroprotective potential of the optimized extract was further evaluated through in vitro inhibition assays targeting acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and lipoxygenase (LOX), alongside a permeability assessment using an in vitro blood–brain barrier (BBB) model. Molecular docking simulations were performed to explore the interactions of apigenin—the most representative flavonoid in the optimal extract—with the three target enzymes. Overall, these findings support the valorization of ABP as a source of bioactive compounds and highlight the potential of PLE as an efficient and sustainable extraction approach. Full article

Background: Hepatic ischemia–reperfusion injury (IRI) remains a major challenge in liver transplantation (LTx) and hepatectomy. Previous studies identified a 12/15-lipoxygenase (12/15-LOX)-driven lipid peroxidation cascade promoting cell death, whereas glutathione peroxidase 4 (GPx4)-dependent metabolism acts antagonistically. This study investigated whether tacrolimus protects against hepatic IRI through this redox axis. Methods: Male C57BL/6 mice underwent 65% partial hepatic warm ischemia and reperfusion with or without tacrolimus preconditioning. Liver tissue and serum were analyzed by spectral photometry, Western blotting, TUNEL assay, and serum enzyme measurement. Results were statistically analyzed and compared with previously published results of 12/15-LOX inhibition by baicalein pretreatment and its carrier DMSO. Also, the combination of both tacrolimus and baicalein was investigated. Results: Tacrolimus increased the oxidative glutathione activity quotient (GSSG/GSH) by 75.1% (p = 0.0302), attenuated MAPK signaling, reduced SAPK/JNK by 84.6% (p = 0.0059), with ERK1/2 showing a downward trend, decreased Caspase-3 activation by 66.9% (p < 0.001) and PARP cleavage by 59.9% (p = 0.0330), and lowered TUNEL-positive cell death by 61.8% (p = 0.0015). Tacrolimus achieved hepatoprotection comparable to 12/15-LOX inhibition, but without hepatotoxicity, whereas combined treatment conferred no additional benefit yet bore toxic properties. Conclusions: Tacrolimus preconditioning mitigates hepatic IRI through a glutathione-linked redox–signaling–cell death axis and exerts cytoprotective effects beyond immunosuppression. Full article

Dietary arachidonic acid (ARA) is essential for aquatic animal growth and health, but studies in bivalves are still limited. Here, microcapsule diets with increasing ARA concentrations (ARA1-6 groups: 0.35, 3.01, 5.25, 6.88, 8.69, and 10.27 mg g⁻¹ dry matter) were prepared by spray drying, and clam Sinonovacula constricta juveniles were fed these diets for 14 days. Results showed that dietary ARA concentrations did not significantly affect clams’ survival, weight gain, and shell length gain rates. The clams in the ARA6 group had significantly higher crude lipid content than those in the other microcapsule groups. The ARA concentrations in the clams increased with higher dietary ARA, while n-3 polyunsaturated fatty acids (PUFAs) and eicosapentaenoic acid (EPA) concentrations decreased. The mRNA levels of cyclooxygenase 2 and 5-lipoxygenase type 2 were significantly higher in the ARA5 and ARA6 groups compared to the ARA1 group. The mRNA levels of 5-lipoxygenase type 3, toll-like receptor 4, and nuclear factor-kappa b p50 (nfκb p50) were significantly higher in the ARA6 group compared to the ARA1 group. As dietary ARA concentrations increased, the mRNA levels of glutamate–cysteine ligase catalytic subunit and glutathione S-transferase, along with malondialdehyde (MDA) content, increased in the clams. Additionally, the superoxide dismutase and catalase activities in the ARA5 and ARA6 groups were significantly higher than those in the ARA1 and ARA2 groups. Clam ARA content, acting as a central node, showed very strong positive correlations with MDA and cyclooxygenase 2, and very strong negative correlations with EPA and the n-3/n-6 PUFA ratio. Our results revealed that high dietary ARA, while not affecting growth, reduced the n-3/n-6 PUFA ratio and induced a response characterized by the upregulation of NF-κB and Nrf2 pathway genes in S. constricta. Full article

Background: Long-term diabetes mellitus may precipitate severe complications, including cognitive dysfunction. Existing research has shown that diabetic cognitive impairment (DCI) in rats is characterized by memory deterioration and a disordered arrangement of hippocampal cells. The Shichangpu–Xiyangshen herb pair (SX) effectively improved the pathological changes induced by DCI. However, the role of SX in regulating the physiological and behavioral responses to DCI remains unclear. Methods: We sought to determine the small-molecule metabolites of cerebrospinal fluid (CSF) and delineate the pathways to elucidate the potential mechanism of the effect of SX in the treatment of DCI by metabolomics strategies, focusing on key mechanisms. Behavioral assessments were conducted on DCI rats and the rats treated with SX, as well as an evaluation of neuronal morphology in the hippocampal region. Metabolomics was used to analyze biomarkers in cerebrospinal fluid at different time points during the development of DCI, to uncover the underlying core mechanisms of DCI, and to investigate the regulatory effects of SX on these core mechanisms. The mechanisms of SX on DCI were investigated using quantitative reverse transcription polymerase chain reaction, immunohistochemistry, Western blot, and ELISA. Results: The Morris water maze (MWM) and social interaction test results revealed that SX administration effectively counteracted cognitive impairments in rats with DCI while simultaneously diminishing pathological damage in the CA1, CA3, and DG hippocampal regions. Further analysis showed that SX restored the significantly reduced levels of IL-8, ROX, and TNF-α, and reduced Aβ plaque formation (as indicated by APP and BACE1 protein expression). Simultaneously, SX markedly ameliorated arachidonic acid metabolic disorders in DCI, including significant reductions in arachidonic acid (AA), PGE2, and LTB4 and reduced expression of COX-2 (PTGS2) and 5-LOX (ALOX-5). Conclusions: Our findings indicate that SX effectively counteracted cognitive impairment in rats with DCI by inhibiting AA metabolism through both cyclooxygenase and lipoxygenase pathways, thereby minimizing neuronal damage. Full article

Alterations in sphingolipids (SLs), oxylipins (OLs) and cytokines (CKs) are central to neuroinflammation. However, the effects of low doses Fumonisins B (FBs) on these analytes in the avian brain remain unclear.This study investigated SLs, OLs, CKs, and the activities of phospholipase A2c (PLA2c) and cyclooxygenase 2 (COX2) in the brains of chickens exposed to FB at a nominally safe dose of 14.6 mg FB1 + FB2/kg for 14 and 21 days. Targeted LC-MS/MS analyses revealed that FB exposure increased brain concentrations of sphingosine, N-acetyl-sphingosine, sphingosine 1-phosphate (So1P), ceramides (Cers), and sphingomyelins (SM). The Cer:SM ratio was elevated at 14 days but normalized by 21 days, whereas the So1P:Cer ratio rose at 14 days and continued to increase at 21 days. These changes coincided with elevated PLA2c and COX2 activities. OL profiling indicated a modest rise in pro-inflammatory arachidonic acid-derived COX metabolites at 14 days, while anti-inflammatory OLs derived from COX and lipoxygenase (LOX) pathways, including PGE2, 15-HETE, and 17-HDHA, increased significantly at 21 days. In contrast, the levels of CKs changed only slightly. Brain concentrations of Fumonisin B1 (FB1) indicated increased blood–brain barrier permeability.These findings highlight a key role of Cers in modulating OL production in FB neurotoxicity. Full article