Search Results (391)

Ferroptosis, a type of iron overload-induced cell death, is involved in diabetes-induced testicular dysfunction. Hence, this study was designed to investigate, for the first time, the impact of lipoxin A4 (LXA4) administration on testicular tissue in diabetic rats and explore its probable role in regulating ferroptosis in comparison with the standard ferroptosis inhibitor (ferrostatin-1, Fer-1). Albino rats of Wistar strain were divided into a control group, a type II diabetes mellitus (DM) group, a DM + Fer-1group, and a DM + LXA4 group. Serum levels of iron, insulin, glucose, total cholesterol, triglycerides, and testosterone were assayed. Testicular tissue markers of oxidative stress, ferroptosis, and inflammation were also assessed by different methods. Our results confirmed diabetes-induced testicular injury and disruption of its function via inducement of ferroptosis, but this was ameliorated with LXA4 and Fer-1 administration. However, Fer-1 showed a greater protective effect compared to LXA4 under the conditions of this study. We concluded that LXA4 partially secured the testicular tissue of diabetic rats against ferroptosis via augmenting the antioxidant Nrf2/SLC7A11/GPX4 pathway. Therefore, LXA4 may have a possible protective effect on the testicular tissue of diabetic patients. Full article

Idiopathic pulmonary fibrosis (IPF) is characterised by progressive parenchymal remodelling, driven by epithelial dysfunction, fibroblast activation, and altered immune regulation within the distal lung. Alveolar macrophages (AMs) reside in a surfactant-rich environment and are specialised for continuous lipid handling, yet the significance of this metabolic role for macrophage heterogeneity and fibrotic progression has remained incompletely integrated across studies. In this review, we synthesise evidence from human lung tissue, experimental models, lipidomic analyses, and clinical investigations to place macrophage populations described in IPF—including FABP4-high homeostatic cells and SPP1-associated disease-enriched states—within a unified lipid-metabolic context. We show that macrophage heterogeneity in IPF can be understood as a variation within a core lipid-handling programme rather than the emergence of distinct macrophage lineages. Profibrotic macrophage states are characterised by altered lipid processing and signalling, including dysregulated sterol handling, lysophospholipid pathways, and eicosanoid balance, which impair surfactant turnover and contribute to fibroblast activation. Importantly, experimental and clinical data indicate that macrophage lipid-metabolic programmes remain modifiable, although definitive disease-modifying efficacy in IPF has yet to be established. Framing macrophage states within a lipid-metabolic framework provides a coherent basis for interpreting heterogeneous datasets and supports the rationale for therapeutic strategies aimed at stabilising or restoring macrophage lipid handling in fibrotic lung disease. Full article

Human infertility affects approximately 17.5% of the global population, with male factors accounting for nearly half of all cases. Identifying reliable molecular biomarkers is crucial for improving the diagnosis and assessment of male fertility. This study established and refined an untargeted high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (HPLC-ESI-MS/MS) protocol for a comprehensive lipidomic and metabolomic analysis of human spermatozoa, using only 1.25 million cells per sample. Compared with previous reports, our optimized method achieved an unparalleled level of analytical depth, identifying 473 lipid species and 955 structurally annotated metabolites. This corresponds to nearly a 7600-fold improvement in detection efficiency per cell compared with previously published approaches. Lipidomic analysis revealed that the most abundant lipid classes were glycerophospholipids (39%), cholesterol (20%) and fatty acids (19%), with cholesterol representing the single most abundant compound. This observation is consistent with the structural complexity of the sperm plasma membrane. Metabolomic profiling similarly identified glycerophospholipids (44%), eicosanoids (14%) and N-acyl amino acids (12%) as the major metabolite classes. The integration of lipidomic and metabolomic data highlighted functionally interconnected pathways related to membrane dynamics, energy metabolism, and hormone biosynthesis. Overall, this work establishes a robust, sensitive, and scalable analytical framework that enables the high-coverage molecular characterization of spermatozoa from limited sample material, laying the groundwork for future biomarker discovery and clinical applications in male infertility research. Full article

Extracellular vesicles (EVs), particularly exosomes, have emerged as critical mediators of intercellular communication, yet the metabolite fraction of their cargo remains substantially underexplored relative to proteins and nucleic acids. This review synthesizes current knowledge on the exosomal metabolome as a functionally distinct intercellular signaling system with unique biophysical properties. We review the mechanisms proposed to govern metabolite encapsulation into exosomes, encompassing membrane transporter involvement, lipid raft partitioning, and binding to luminal proteins, and discuss the unresolved question of whether metabolite loading is selective or stochastic. Critically, we present a quantitative framework evaluating whether delivered metabolite quantities are sufficient to alter recipient cell metabolic pools, distinguishing receptor-mediated signaling from bulk substrate delivery. We also address methodological considerations including contamination artifacts and isolation-method biases that complicate interpretation of EV metabolomics data. Exosomal metabolites are reviewed across four functional categories: energy substrates (ATP, lactate, amino acids), signaling molecules (TCA cycle intermediates, eicosanoids, nucleotides), redox cofactors and antioxidants (NADH, glutathione), and oncometabolites. For each category, available evidence is critically appraised, distinguishing metabolites with direct mass spectrometric detection from those whose roles are inferred from parent-cell biology. The review examines the roles of exosomal metabolites in tumor-stroma metabolic symbiosis, immunometabolic regulation, inter-organ crosstalk in metabolic diseases including type 2 diabetes and non-alcoholic fatty liver disease, cancer metastasis, viral infections, and immune evasion. A quantitative framework is discussed to evaluate whether delivered metabolite quantities are sufficient to alter recipient cell metabolic pools, distinguishing receptor-mediated signaling from bulk substrate delivery. Technical challenges in exosomal metabolomics are reviewed, including the impact of isolation method on data quality, contamination artifacts, and current standardization gaps. Therapeutic implications of exosomal metabolite signaling are discussed, encompassing metabolite-loaded exosomes as therapeutic vehicles and exosomal metabolite loading as a pharmacological target. Integration of single-vesicle technologies with systems biology approaches is highlighted as a promising direction for advancing this field toward precision medicine applications in oncological and metabolic disorders. Full article

Mammalian arachidonic acid lipoxygenases (ALOXs) are lipid-peroxidizing enzymes, which have been implicated in inflammatory, hyperproliferative and neurodegenerative diseases. Nordihydroguaiaretic acid (NDGA) is a naturally occurring antioxidant and a potent lipoxygenase inhibitor. Unfortunately, the molecular basis of the NDGA–ALOX interaction remains unexplored. Here, we show by in silico docking studies and by molecular dynamics simulations that NDGA binds in the substrate binding pocket of human ALOX15 and that Gln595 plays a major role in this interaction. In silico mutagenesis studies (Glu595Ala, Glu595Leu, Glu595Glu, Glu595Ile) modified the stability of the ALOX15–NDGA complex and altered the ligand binding behavior of the enzyme. To validate the in silico findings, we expressed human ALOX15 and the enzyme mutants as recombinant proteins, characterized their functional properties and quantified the IC₅₀ values for NDGA-induced inhibition. Consistent with our in silico predictions, the experimental IC₅₀ values demonstrated that NDGA strongly inhibited wildtype ALOX15 and its Gln595Glu and Gln595Ile mutants. In contrast, the IC₅₀ values for the Gln595Ala and Gln595Leu mutants were more than one order of magnitude higher. These findings highlight the role of Gln595 for the NDGA–ALOX15 interaction and may facilitate the future development of isoform-specific ALOX15 inhibitors. Full article

Chronic obstructive pulmonary disease (COPD) is a leading cause of mortality worldwide, yet only a fraction of smokers develops the disease, suggesting protective mechanisms in resilient individuals. Identifying airway-localized molecular signatures may improve our understanding of disease pathomechanisms and support hypothesis generation for biomarker research. In this pilot study, induced sputum from smokers with COPD (n = 28) and smokers without COPD (n = 16; Global Initiative for Chronic Obstructive Lung Disease (GOLD)-defined pre-COPD) was analyzed by untargeted proteomics, metabolomics, and lipidomics. After quality control, 1180 proteins, 187 metabolites, and 1234 lipids were retained. Analyses included univariate models with false discovery rate adjustment and multivariate analyses (PCA, PLS-DA), followed by pathway enrichment and protein interaction network analysis. While few features remained significant after FDR correction, consistent cross-omics patterns were observed. COPD was characterized by ↑ glutathione, creatine, and L-arginine; ↓ CCDC88A and ↑ STAT3 and SYDE2; and broad lipid remodeling involving phosphatidylcholines, sphingolipids, and eicosanoids. Network analysis highlighted STAT3 as a highly connected node linking COPD-related genes. These findings suggest that the multi-omic profiling of induced sputum can capture coherent airway-localized molecular signatures such as oxidative stress, cytoskeletal remodeling, and Rho-family GTPase signaling. However, the results should be interpreted as exploratory and require validation in functional studies. Full article

The replacement of fishmeal with plant protein is widely regarded as a key strategy for sustainable aquaculture. However, carnivorous marine fish often show limited tolerance to fishmeal-free diets. Here, we investigated growth performance, hepatic physiological responses, and molecular mechanisms underlying adaptation to a soy protein concentrate-based diet (SPCD) in golden pompano (Trachinotus ovatus). An 8-week feeding trial was conducted under communal rearing conditions, followed by the phenotypic stratification of SPCD-fed fish into high- and low-growth subgroups. Growth performance, serum biochemical indices, and liver histology were assessed, and integrated transcriptomic and metabolomic analyses were performed on liver tissue. At the population level, the SPCD resulted in reduced growth, a lower feed intake, and decreased feed utilization efficiency compared with a fishmeal-based diet. Notably, marked inter-individual variation was observed: fish fed the SPCD exhibited significantly lower final body weights and a higher FCR compared with the FMD group (p < 0.001), and pronounced growth divergence was observed between the PB and PS subgroups, with a subset of SPCD-fed fish maintaining growth comparable to fishmeal-fed controls, whereas others exhibited severely constrained growth. Divergent phenotypes were associated with distinct hepatic alterations, including aggravated vacuolation, the enrichment of tight junction-related and immune regulatory pathways, and the broad reprogramming of lipid metabolism. Integrated multi-omics analysis identified arachidonic acid metabolism as the most significantly perturbed pathway, characterized by altered membrane phospholipid composition, the upregulation of RARRES3L, increased COX/LOX-derived eicosanoids, and the suppression of the CYP–EET branch. Collectively, these findings indicate that soy protein replacement induces coordinated hepatic structural and metabolic remodeling, with tight junction disruption and arachidonic acid metabolic reprogramming contributing to inflammatory imbalance and divergent growth phenotypes in T. ovatus. Full article

The optimal dietary balance between n-6 and n-3 polyunsaturated fatty acids (PUFAs), the safe upper intake of n-6 PUFAs—particularly linoleic acid—and the physiological consequences of their metabolic competition remain unresolved in the context of the Western diet. Since the 1980s, Bill Lands and colleagues have argued that high n-6 PUFA intake can shift the balance of n-3-derived pathways and eicosanoid signaling, potentially influencing processes relevant to non-communicable diseases. Across human populations, the proportion of n-6 in tissue HUFA spans a broad range—from roughly 20% in traditional dietary patterns to nearly 80% in typical Western diets—illustrating the predictable impact of dietary precursor supply on HUFA composition. Despite its potential public health implications, this hypothesis has received limited systematic attention. In this narrative review, we synthesize key aspects of Lands’ work, evaluate supportive and contradictory evidence, and highlight mechanistic insights into lipid competition and inflammatory regulation. We conclude that these unresolved but testable hypotheses warrant renewed investigation, as their corroboration could reshape dietary guidelines and strategies for chronic disease prevention. Full article

Colorectal cancer (CRC) is a major health concern with increasing incidence, especially in younger adults. This study evaluated the stage-dependent role of serum eicosanoids as biomarkers in CRC patients. A cohort of 122 patients undergoing curative colorectal resection was prospectively recruited. Serum eicosanoid profiles were evaluated using targeted metabolomics and analyzed through regression-based statistical models to identify associations with CRC staging. The more advanced stages of CRC (with N+ and M+) showed significantly increased levels of PGD₂, PGE₂, and TXB₂. The latter proved to be consistently associated with advanced disease. LTB₄ and PGD₂ showed inverse relationships relative to each other with respect to local invasion, showing PGD₂ as a marker of higher T stages. PGE₂ was not recognized as a viable biomarker. The progression of CRC is associated with distinct alterations in eicosanoid profiles. This study showed the potential of TXB₂, LTB₄, and PGD₂ as indicators of CRC advancement. Full article

Background: Caloric restriction (CR) is a dietary intervention based on limiting calories relative to the basic energy needs of the organism, which changes the intensity of metabolism, causes changes in the functioning of the endocrine and sympathetic systems, and influences the expression of genes in muscle, heart, and brain cells. During the use of CR, there is a transition from carbohydrate supply to increased fat metabolism. Fatty acids are more or less susceptible to free radicals, depending on their molecular structure. Oxidation (peroxidation) contributes to the production of metabolites (including hydroxyeicosatetraenoic acid and hydroxyoctadecadienoic acid), some of which are involved in inflammation. Methods: The aim of this study was to evaluate the effects of long-term caloric restriction on the tissue levels of selected fatty acids and fatty acid-derived lipid mediators with pro-inflammatory or anti-inflammatory properties in skeletal muscle and intestinal tissues. The study was carried out on C57BL/6 mice. During the 8-month experiment, the mice in the study group were fed a 30% calorie restricted diet—according to the Every-Other-Day Diet concept. Analyses were performed on intestinal and muscle tissues collected from animals. Fatty acid derivatives were isolated using solid-phase extraction (C-18 columns) columns, and isolation of fatty acids was performed using a modified Folch method. The compounds were analyzed by liquid and gas chromatography. Results: CR induced detectable alterations in both fatty acid profiles and lipid mediator concentrations in a tissue-specific manner. However, most of these changes did not remain statistically significant after multiple testing correction. Conclusions: These findings suggest potential effects of long-term CR on lipid signaling pathways, although the current dataset lacks the statistical power required to draw definitive conclusions. This study highlights the need for further research using larger sample sizes and integrated multiomic approaches to elucidate the molecular mechanisms underlying lipidomic adaptations to prolonged caloric restriction. Full article

Bisphenol AF (BPAF), a prevalent bisphenol A (BPA) substitute, raises concerns due to its environmental persistence and endocrine-disrupting potency. While metabolic effects of direct exposure are documented, its intergenerational consequences remain unclear. Here, we demonstrated that perinatal BPAF exposure induced persistent metabolic syndrome in offspring, including glucose intolerance, hepatic steatosis, and adipose hypotrophy. Integrating multi-omics data, we observed that BPAF exposure shaped offspring’s hepatic epigenome, as demonstrated by genome-wide alterations in H3K27ac-marked regulatory elements. This epigenetic rewiring indicated a dual regulatory effect on transcriptomes that suppressed interferon-γ responses while activating sterol biosynthesis, ultimately perturbating hepatic metabolome, including depleted pantothenate levels and accumulation of pro-inflammatory eicosanoids. Our findings suggest that BPAF may act as a developmental toxicant capable of persistently disrupting the immune–metabolic axis through epigenomic mechanisms, highlighting the need for careful re-evaluation of its use as a BPA substitute in consumer products. Full article

The COVID-19 pandemic exposed critical vulnerabilities in single-target antiviral strategies, highlighting the urgent need for multi-mechanism therapeutic approaches against emerging viral threats. Here, we present an integrated computational framework systematically evaluating natural fatty acids as potential dual ACE2 (Angiotension Converting Enzyme 2)-inflammatory modulators; compounds simultaneously disrupting SARS-CoV-2 viral entry through allosteric ACE2 binding while suppressing host inflammatory cascades; through allosteric binding mechanisms rather than conventional competitive inhibition. Using molecular docking across eight ACE2 regions, 100 ns molecular dynamics simulations, MM/PBSA free energy calculations, and multivariate statistical analysis (PCA/LDA), we computationally assessed nine naturally occurring fatty acids representing saturated, monounsaturated, and polyunsaturated classes. Hierarchical dynamics analysis identified three distinct binding regimes spanning fast (τ < 50 ns) to slow (τ > 150 ns) timescales, with unsaturated fatty acids demonstrating superior binding affinities (ΔG = −6.85 ± 0.27 kcal/mol vs. −6.65 ± 0.25 kcal/mol for saturated analogs, p = 0.002). Arachidonic acid achieved optimal SwissDock affinity (−7.28 kcal/mol), while oleic acid exhibited top-ranked predicted binding affinity within the computational hierarchy (ΔG_bind = −24.12 ± 7.42 kcal/mol), establishing relative prioritization for experimental validation rather than absolute affinity quantification. Energetic decomposition identified van der Waals interactions as primary binding drivers (65–80% contribution), complemented by hydrogen bonds as transient directional anchors. Comprehensive ADMET profiling predicted favorable safety profiles compared to synthetic antivirals, with ω-3 fatty acids showing minimal nephrotoxicity risks while maintaining excellent intestinal absorption (>91%). Multi-platform bioactivity analysis identified convergent anti-inflammatory mechanisms through eicosanoid pathway modulation and kinase inhibition. This computational investigation positions natural fatty acids as promising candidates for experimental validation in next-generation pandemic preparedness strategies, integrating potential therapeutic efficacy with sustainable sourcing. The framework is generalizable to fatty acids from diverse biological origins. Full article

Vascular endothelium balances antithrombotic and anti-inflammatory activity to control blood vessel tone under physiological conditions. However, endothelial dysfunction impairs these processes, causing a state that promotes clotting and inflammation. Eicosanoids are a major class of bioactive lipid mediators crucial for modulating endothelial and platelet function. Research has highlighted the roles of eicosanoids in vascular diseases, showing pro-inflammatory, prothrombotic, and protective activities. Specifically, prostaglandin E₂ (PGE₂) is crucial because of its major role in atherosclerosis development and progression, acting via EP receptors involved in forming, maintaining, and stabilizing atherosclerotic lesions, thereby making PGE₂-EP signalling a specific target for treating cardiovascular diseases. This review will explore the evidence on eicosanoids and the role of their receptor modulation in platelet and vascular dysfunction in atherothrombosis. The studies included in this scoping review were retrieved from PubMed, Web of Science, Cochrane, and Scopus in accordance with the Preferred Reporting Items for Scoping Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) statement and the Population Intervention Comparison Outcome Population (PICO) framework. Eight clinical studies were found, which highlighted the crucial role of eicosanoids, like prostaglandins and their receptors, in endothelial and platelet dysfunction, and also how pharmacological mechanisms affect atherothrombosis. A new therapeutic approach for cardiovascular dysfunction is indicated by the recent findings, specifically against atherothrombosis, focusing on eicosanoids, their receptors, and processes like oxidative stress. Despite this evidence, there is a lack of comprehensive research results from scientific databases; therefore, further in vitro, in vivo, and clinical studies should be promoted to validate the preliminary results. Full article

Background: Chronic inflammation contributes to functional decline in older adults, yet interventions targeting inflammatory pathways have shown inconsistent results. Metabolomics offers a promising approach to identify biological heterogeneity and uncover molecular signatures underlying differential functional trajectories. Objective: Our objective was to examine whether untargeted serum metabolomics can identify metabolic signatures associated with baseline physical function, functional trajectories, and treatment response in older adults with chronic inflammation participating in the ENRGISE trial. Methods: We performed untargeted metabolomic profiling on serum samples (n = 731) collected at baseline, 6, and 12 months from participants (mean age ≥ 70) enrolled in the ENRGISE pilot randomized trial. Participants were randomized to losartan, omega-3 supplementation, both, or placebo. Functional measures included grip strength and 400 m gait speed. Group-based trajectory modeling classified participants into functional trajectories over 12 months. Partial least squares-discriminant analysis (PLS-DA) and pathway enrichment (mummichog algorithm) were used to identify differentially abundant metabolites and perturbed pathways. Results: Baseline metabolomic profiles differed by physical function status. Participants with low grip strength showed enrichment in vitamin A metabolism pathways, while slower gait speed was associated with higher levels of prostaglandin and eicosanoid metabolites. Baseline metabolic profiles distinguished individuals who later declined versus improved in functional performance. Omega-3 supplementation, but not losartan, induced distinct changes in lipid-related pathways, including fatty acid activation, omega-3 metabolism, and prostaglandin biosynthesis, indicating that individuals responded to these interventions metabolically despite null clinical outcomes. Conclusions: Serum metabolomic signatures were associated with baseline physical function, predicted functional trajectories, and revealed pharmacologic activity of omega-3 supplementation. These findings support the use of metabolomics to uncover biological heterogeneity and inform precision geroscience strategies in aging populations. Full article

Background/Objectives: Different oxylipin subtypes have unique biological properties, requiring effective analytical protocols. However, establishing a complete pathway detection protocol for comprehensive oxylipin analysis is challenging. This study aimed to evaluate the adaptability and specificity of oxylipin subtypes under different extraction schemes and to develop a robust analytical platform for clinical biomarker investigation. Methods: We revealed the adaptability and specificity of oxylipin subtypes based on different single-step extraction schemes. A high-throughput quantitative automated solid-phase extraction coupled with a liquid chromatography–tandem mass spectrometry (aSPE–LC–MS/MS) analytical platform was established for a broad panel of complex oxylipins. The method was applied to serum samples of patients with coronary heart disease (CHD). Results: Our results verified that oxo-oxylipins, resolvin, and eicosanoids showed the best extraction efficiency under SPE protocol. Most hydroxy-oxylipins, dihydroxy-oxylipins, and HOTrEs are suitable for methanol protocol, HDHA for acetonitrile protocol, and epoxy-oxylipins for the methyl tert-butyl ether protocol, while medium-chain HETE is suitable for ethyl acetate protocol. Importantly, a novel sensitive fast method with wide coverage by the aSPE–LC–MS/MS analytical platform with satisfying sensitivity, accuracy and precision, extraction efficiency, low matrix effect, and linear calibration curves was obtained. Furthermore, we have successfully applied this method and found that 5-HETE, 11-HETE, and 15-HETE can serve as integrated biomarkers for patients with CHD, with high diagnostic performance. Conclusions: The study provides the best protocol for the clinically targeted detection of oxylipins and provides an important means for studying biomarkers of diseases. Full article