Metabolites

27 pages, 3319 KB

Open AccessArticle

Screening of “Cry for Help” Signals from Angelica sinensis Induced by Fusarium solani and Their Potential for Biological Control

by Tianpeng Xie, Qi Ding, Linhua Yang, Jingyi Wang, Jingxian Wei, Xiaoxue Du and Ling Jin

Metabolites 2026, 16(6), 385; https://doi.org/10.3390/metabo16060385 - 2 Jun 2026

Abstract

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 [...] Read more.

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

(This article belongs to the Special Issue Metabolomics and Plant Defence, 2nd Edition)

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18 pages, 1108 KB

Open AccessArticle

Effect of Icosa-11,14,17-Trienoic Acid from Setipinna phasa Oil on Lipogenesis and Adipose Inflammation on Mice with High Fat Diet Induced Obesity

by Titli Panchali, Riya Kar, Pipika Das, Ananya Dutta, Manisha Phoujdar, Kuntal Ghosh and Shrabani Pradhan

Metabolites 2026, 16(6), 384; https://doi.org/10.3390/metabo16060384 - 1 Jun 2026

Abstract

Background/Objectives: Obesity is a complex disease involving the accumulation of an excessive amount of body fat. It is a condition that develops when energy intake and expenditure are out of balance. Inflammation and hypertrophy are caused by the storage of too much [...] Read more.

Background/Objectives: Obesity is a complex disease involving the accumulation of an excessive amount of body fat. It is a condition that develops when energy intake and expenditure are out of balance. Inflammation and hypertrophy are caused by the storage of too much white adipose tissue, resulting in adiposity, which also secretes several pro-inflammatory cytokines. Several marketed drugs used to treat obesity have many side effects from long-term ingestion. Other therapeutic compounds from marine sources have already been established for treating obesity. In this paper, the main aim is to establish the anti-obesity effect of derived omega-3 fatty acids, i.e., 20:3(n-3)11-14-17 Icosa Trienoic Acid from Setipinna phasa oil. Methods: In the present investigation, inbred male Swiss albino mice were segregated into six categories as Control, Positive Control, Obese Control, and 20:3(n-3)11-14-17 Icosa Trienoic Acid treated groups with three different doses: Treatment 1, Treatment 2 and Treatment 3. To establish the potentiality of extracted fatty acid, different parameters would be considered, such as body weight, lipid composition and different obesity and obesity-associated inflammation markers. Results: After the isolated compound from Setipinna phasa oil was applied to the treated mice group, it decreased their body weight and serum lipid profile by 39.05%, 62.69%, 62.72%, and 78.46% compared to obese mice. They also had lower levels of uric acid, Serum Glutamic-Oxaloacetic Transaminase, Serum Glutamic Pyruvic Transaminase, and Alkaline Phosphatase, at 67.52%, 57.09%, 64.80%, and 43.99%, than the obese group. Accordingly, the treated group’s expression of genes linked to obesity and pro-inflammatory cytokines was downregulated. The isolated compound affected both anti-inflammatory and anti-obesity markers’ increased expression. Conclusions: After the experiments, it was found that the possibility of using fatty acids might be helpful as an anti-inflammatory and anti-obesity therapeutic strategy. This therapeutic strategy will be cheap and cost-effective. Full article

(This article belongs to the Topic Natural Products in Prevention and Therapy of Metabolic Syndrome)

19 pages, 3861 KB

Open AccessReview

Circadian Regulation of Gut Microbial Metabolites in Intestinal Epithelial Homeostasis

by Miri Park, Sooah Kim and Jeehwan Choe

Metabolites 2026, 16(6), 383; https://doi.org/10.3390/metabo16060383 - 1 Jun 2026

Abstract

The gut microbiota produces chemically diverse metabolites whose levels fluctuate depending on the time of day, driven by bidirectional coupling between host intestinal circadian clocks and intrinsic microbial oscillators. Although short-chain fatty acids have received the most attention as microbial circadian effectors, a [...] Read more.

The gut microbiota produces chemically diverse metabolites whose levels fluctuate depending on the time of day, driven by bidirectional coupling between host intestinal circadian clocks and intrinsic microbial oscillators. Although short-chain fatty acids have received the most attention as microbial circadian effectors, a broad class of metabolites, including secondary bile acids, indole derivatives, and branched-chain fatty acids, engage distinct epithelial receptors and transcriptional programs through mechanisms that are, to varying degrees, subject to circadian regulation. However, the mechanisms by which these metabolite classes collectively regulate barrier integrity, mucosal immune tone, and stem cell-driven renewal, as well as the consequences of their rhythmicity loss under circadian misalignment, have not been systematically reviewed. This review constructs a mechanistic framework linking microbial metabolite rhythmicity to the circadian regulation of intestinal epithelial homeostasis and evaluates dietary and probiotic interventions that modulate this axis as chronobiotic strategies. Convergent mechanisms, unresolved questions, and translational opportunities are identified across in vitro, preclinical, and clinical evidence. Full article

(This article belongs to the Special Issue Modulating Gut Microbiota and Host Interactions: The Role of Diet, Probiotics, and Circadian Rhythms)

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13 pages, 1179 KB

Open AccessArticle

Combinatorial Effects of Boron Compounds on Immunophenotypic Modulation of Mouse Hematopoietic Stem Cell Subsets Ex Vivo

by Fatih Kocabaş, Eray Esendir and Neslihan Meriç

Metabolites 2026, 16(6), 382; https://doi.org/10.3390/metabo16060382 - 31 May 2026

Abstract

Background/Objectives: Hematopoietic stem cells (HSCs) sustain lifelong blood cell production and hold therapeutic promise, yet their ex vivo expansion remains constrained by an incomplete understanding of the metabolic and cellular mechanisms governing self-renewal. In this study, we investigated whether boron compounds boric acid [...] Read more.

Background/Objectives: Hematopoietic stem cells (HSCs) sustain lifelong blood cell production and hold therapeutic promise, yet their ex vivo expansion remains constrained by an incomplete understanding of the metabolic and cellular mechanisms governing self-renewal. In this study, we investigated whether boron compounds boric acid (BA), sodium pentaborate pentahydrate (NaB), and sodium 2-pentaborate pentahydrate-8 (Na2B8) can promote the expansion of mouse HSCs by modulating key stem cell populations linked to metabolic fitness. Methods: Lineage-negative (Lin-) cells were magnetically isolated and treated with each boron compound for four days, followed by flow cytometric analysis of c-Kit, Sca-1, Lin-c-Kit+Sca-1+ (LSK), and LSKCD34^low HSC-enriched subsets. Results: Our results show that boron derivatives exert distinct effects on these cellular markers. Notably, NaB treatment significantly increased the Lin-Sca-1+ cell fraction, while Na2B8 elevated both LSK and LSKCD34^low ratios. Furthermore, the BA+NaB combination produced a statistically significant proliferative effect on Sca-1+ and c-Kit+ (CD117) cells. Conclusions: These findings indicate that specific boron compounds enhance ex vivo HSC expansion through yet-to-be-defined mechanisms that underpin HSC self-renewal. Further mechanistic studies are warranted to delineate the precise metabolic targets, but these results highlight boron compounds as promising tools for improving HSC expansion strategies. Full article

(This article belongs to the Section Cell Metabolism)

16 pages, 1558 KB

Open AccessArticle

Chronic Caffeine Consumption Prevents Body Weight Gain and Glucose Intolerance in High-Fat Diet-Induced Obesity Mice Model

by Giuseppe Faraco, Natália F. Mendes, Luisa O. Schmitt, Tamires S. Stivanin, Elisa Gaspar, Nicolle Platt, Manuella P. Kaster and Joana M. Gaspar

Metabolites 2026, 16(6), 381; https://doi.org/10.3390/metabo16060381 - 31 May 2026

Abstract

Background/Objectives: Caffeine consumption has been reported to have beneficial effects in metabolic disorders; however, its effects on food intake are not fully elucidated. This study evaluated the impact of chronic caffeine consumption on weight gain, food intake, and metabolic parameters in C57BL/6 [...] Read more.

Background/Objectives: Caffeine consumption has been reported to have beneficial effects in metabolic disorders; however, its effects on food intake are not fully elucidated. This study evaluated the impact of chronic caffeine consumption on weight gain, food intake, and metabolic parameters in C57BL/6 male mice. Methods: Eight-week-old male mice (28 animals) were divided into four groups: control (chow diet), caffeine (chow diet + 1 g/L caffeine in drinking water), high-fat diet (HFD), and HFD + caffeine (HFD + 1 g/L caffeine in drinking water). Diets and caffeine were provided ad libitum for 8 weeks. Food and water intake were recorded weekly, and blood glucose was measured every 4 weeks. After 8 weeks of diet and caffeine exposure, metabolic tests were conducted, and tissues were collected for biochemical analysis. Results: HFD consumption for 8 weeks induced an increase in body weight and adiposity compared to the chow diet, without changes in food intake. Caffeine consumption prevented body weight gain and adiposity, although it increased food intake. Caffeine also improved glucose tolerance in the HFD mouse model, without changes in random blood glucose, triglyceride, or cholesterol levels. Analysis of hypothalamic neuropeptide (Agrp, NPY, Pomc, Cart), involved in the control of food intake, showed no differences in expression. There were also no changes observed in locomotion nor in anxiety-like behavior. Conclusions: In conclusion, chronic high-fat diet (HFD) exposure induced obesity characterized by increased body weight and adiposity without altering food intake. Chronic caffeine consumption counteracted HFD-induced weight gain and fat accumulation and improved glucose tolerance, despite increasing food intake. Importantly, caffeine consumption in the HFD group did not affect locomotor activity or anxiety-like behavior, suggesting that its metabolic effects are not driven by changes in general activity or emotional state. Overall, these findings indicate that chronic caffeine consumption improves metabolic homeostasis in HFD-fed mice. Full article

(This article belongs to the Special Issue The Interaction Between Metabolic Dysfunction and Hypothalamic Inflammation)

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35 pages, 2332 KB

Open AccessReview

The Application of Metabolomics in Frailty: Trends, Challenges, and Future Directions

by Kaiying Fang, Bei Niu, Zhen Zhang, Yameng Jiang, Ya Zhao and Zhanguo Wang

Metabolites 2026, 16(6), 380; https://doi.org/10.3390/metabo16060380 - 31 May 2026

Abstract

Frailty is a geriatric syndrome involving inflammation, oxidative stress, mitochondrial dysfunction, and metabolic disturbances. Metabolomics can systematically elucidate metabolic pathways and identify actionable biomarkers. This study systematically reviews the progress and evolutionary trends of metabolomics applications in frailty research from 2006 to 2025. [...] Read more.

Frailty is a geriatric syndrome involving inflammation, oxidative stress, mitochondrial dysfunction, and metabolic disturbances. Metabolomics can systematically elucidate metabolic pathways and identify actionable biomarkers. This study systematically reviews the progress and evolutionary trends of metabolomics applications in frailty research from 2006 to 2025. Based on 1924 publications retrieved from the Web of Science Core Collection, systematic analyses were performed using CiteSpace, VOSviewer, SCImago Graphica, and the R package “bibliometrix”, focusing on pathway-level research hotspots and collaboration networks. The United States and China are the leading contributors. Research hotspots have shifted from macro-level biomarkers such as inflammation and protein–energy wasting to specific metabolic pathways including amino acid metabolism, energy metabolism, lipid metabolism, and tryptophan degradation. Key metabolites include sphingomyelin, butyrate, and trimethylamine-N-oxide. Emerging frontiers focus on the association between gut microbiota-derived metabolites and frailty phenotypes, as well as intervention strategies targeting these metabolites. This study provides the first systematic overview of global research progress in metabolomics and frailty, establishes a reproducible evaluation framework integrating physiology, nutrition, geriatrics, and computational biology, and identifies butyrate, trimethylamine-N-oxide, and tryptophan metabolites as potential metabolic targets for early identification and intervention. Full article

(This article belongs to the Section Endocrinology and Clinical Metabolic Research)

16 pages, 1884 KB

Open AccessArticle

Relationships Among Adipose Tissue Distribution, Vitamin D, and Bone Metabolism in Normoglycemic and Type 2 Diabetic Individuals

by Tian-Hang Ma, Juan Zhao, Kun-Hou Zhou, Ya-Xin Guan, Fan Zuo, Xin Nian, Yi Zheng, Wen-Jiao Wang, Li-Juan Zhang, Tsutomu Kazumi, Jingshan Huang and Bin Wu

Metabolites 2026, 16(6), 379; https://doi.org/10.3390/metabo16060379 - 31 May 2026

Abstract

Objectives: To investigate the interplay between adipose distribution, vitamin D metabolites, and bone mineral density (BMD) in Normal Glucose Tolerance (NGT) and type 2 Diabetic (T2DM) individuals. Methods: 167 participants (NGT: 61; T2DM: 106) were enrolled. Serum 25(OH)D, 1,25(OH)₂D₃, [...] Read more.

Objectives: To investigate the interplay between adipose distribution, vitamin D metabolites, and bone mineral density (BMD) in Normal Glucose Tolerance (NGT) and type 2 Diabetic (T2DM) individuals. Methods: 167 participants (NGT: 61; T2DM: 106) were enrolled. Serum 25(OH)D, 1,25(OH)₂D₃, Parathyroid Hormone (PTH), and Ca were quantified. Visceral (VAT) and subcutaneous (SAT) adipose areas were assessed via dual bioelectrical impedance analysis. BMD and body composition were assessed via DXA. Metabolic indices (HOMA-IR, HOMA-β, ISI) were calculated. Results: 1. NGT: 25(OH)D was unrelated to adiposity. Conversely, 1,25(OH)₂D₃ was correlated inversely with VAT, SAT, body mass index (BMI), and Fat Mass Index (FMI), with VAT being the strongest independent predictor. 2. T2DM: High VAT correlated with insulin resistance yet paradoxically higher BMD. 25(OH)D correlated positively with Z-score, while 1,25(OH)₂D₃ correlated negatively with lumbar BMD. 3. VAT exerted a greater influence on insulin resistance than SAT, particularly in T2DM. Conclusions: 1. Visceral adiposity is the primary determinant of active 1,25(OH)₂D₃ metabolism in both NGT and T2DM individuals. 2. 1,25(OH)₂D₃ levels may be more closely associated with adiposity-related metabolic alterations than 25(OH)D. Despite lower 1,25(OH)₂D₃, the positive association between VAT and BMD in T2DM suggests complex mechanisms where visceral fat may paradoxically influence bone metabolism while driving insulin resistance. Full article

(This article belongs to the Special Issue Vitamin D Metabolism: Implications in Metabolic Health and Disease)

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19 pages, 3714 KB

Open AccessArticle

Genetically Informed Single-Cell Analysis Reveals PLXND1 as a Cell-Type-Specific Molecular Switch in MASLD

by Xianyi Ma, Junbo Song, Xin Hong and Zhibin Lin

Metabolites 2026, 16(6), 378; https://doi.org/10.3390/metabo16060378 - 30 May 2026

Abstract

Background/Objectives: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a systemic disorder driven by genetic predisposition, epigenetic programming, metabolic rewiring, and immune dysregulation. Although population genetics and single-cell transcriptomics have advanced our understanding, the multi-omic causal architecture of MASLD at cellular resolution remains poorly [...] Read more.

Background/Objectives: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a systemic disorder driven by genetic predisposition, epigenetic programming, metabolic rewiring, and immune dysregulation. Although population genetics and single-cell transcriptomics have advanced our understanding, the multi-omic causal architecture of MASLD at cellular resolution remains poorly defined. This study aimed to establish an integrative framework linking genetic causality to cell-type-specific tissue dysfunction. Methods: Multi-layered Mendelian randomization (MR) and summary-data-based MR (SMR) across large-scale eQTL and pQTL datasets were applied to prioritize causal genes. Single-cell eQTL-based MR across 14 immune lineages generated cell-type-specific causal hypotheses, which were validated using human hepatic single-cell RNA-sequencing data (GSE136103). Two-step mediation MR quantified upstream epigenetic and downstream metabolic mechanisms. A high-fat diet (HFD)-induced murine model provided organismal validation. Results: Multi-layered MR nominated PLXND1 as a robust causal driver of MASLD. Single-cell eQTL-based MR revealed a functional dichotomy: PLXND1 upregulation in CD8⁺ effector memory T-cells decreased MASLD risk (OR = 0.486, 95% CI: 0.290–0.813, p = 0.006), whereas upregulation in natural killer cells (OR = 1.567, 95% CI: 1.337–1.837, p < 0.001), non-classical monocytes, and dendritic cells increased risk. Human hepatic single-cell transcriptomics confirmed that PLXND1 marks an anti-fibrotic, IFNG-high CD8⁺ T subset and a pro-inflammatory lipid-associated macrophage (LAM) population. Mediation MR identified DNA methylation at cg26767922 and cg08471739 as protective mediators acting predominantly via PLXND1 downregulation (92.39% and 64.50% mediation, respectively), and linked PLXND1 to six circulating metabolites. HFD mice showed significant hepatic PLXND1 upregulation. Conclusions:PLXND1 functions as a lineage-dependent molecular switch in MASLD, validated across genetic, epigenetic, metabolic, and single-cell dimensions. These findings caution against systemic PLXND1 blockade and support precision therapeutic strategies targeting hepatic innate immune cells. Full article

(This article belongs to the Section Endocrinology and Clinical Metabolic Research)

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23 pages, 5648 KB

Open AccessArticle

Application of Plasma Metabolomic Biomarker Panels in Early Diagnosis and Disease Staging of Alzheimer’s Disease

by Jiao Chen, Xuhui Chen, Ting Chen and Jun Hu

Metabolites 2026, 16(6), 377; https://doi.org/10.3390/metabo16060377 - 30 May 2026

Abstract

Background: Previous metabolomics studies on Alzheimer’s disease (AD) have predominantly focused on Western populations, leaving Chinese cohorts and disease stage-specific data largely unexplored. Objectives: To characterize metabolic alterations across different clinical stages of AD in a Chinese population and identify early [...] Read more.

Background: Previous metabolomics studies on Alzheimer’s disease (AD) have predominantly focused on Western populations, leaving Chinese cohorts and disease stage-specific data largely unexplored. Objectives: To characterize metabolic alterations across different clinical stages of AD in a Chinese population and identify early diagnostic biomarkers. Methods: We enrolled 172 participants, including patients with AD, mild cognitive impairment (MCI), subjective cognitive decline (SCD), vascular cognitive impairment (VCI), and healthy controls (HC). Untargeted metabolomics (LC-MS and GC-MS) was performed on plasma samples, integrated with Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) assessments. Data were analyzed using multivariate statistics, pathway enrichment, and ROC modeling. Results: Distinct metabolic profiles emerged across disease stages, with phospholipids, ceramides, and glucose metabolites prominently enriched in glycerophospholipid, sphingolipid, and glucose pathways. A 16-metabolite panel achieved robust discrimination between AD+MCI and HC+VCI+SCD (AUC = 0.804). Specific metabolites, including ceramides, dihydroceramides, phosphatidylinositol, phosphatidylcholine, and glycodeoxycholic acid, correlated significantly with cognitive function and disease progression. Conclusions: This study reveals stage-specific metabolic dysregulation in Chinese AD patients and identifies potential plasma biomarkers for early detection, offering insights into AD pathogenesis. Trial registration number ChiCTR2400092653. Full article

(This article belongs to the Section Endocrinology and Clinical Metabolic Research)

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18 pages, 13163 KB

Open AccessArticle

Dendrobium huoshanense Ameliorates Sleep Deprivation-Induced Ileal Mucus Barrier Dysfunction by Regulating Steroid Hormone Biosynthesis and the HPA Axis in Rats

by Xue Luo, Shuxiang Jin, Yue Fang, Qun Zhao, Huiqun Xie and Lan Han

Metabolites 2026, 16(6), 376; https://doi.org/10.3390/metabo16060376 - 30 May 2026

Abstract

Background/Objectives: Sleep deprivation (SD) induces the accumulation of reactive oxygen species (ROS) in the intestine, causing inflammation in the intestine, thereby damaging the intestinal epithelial barrier function. As a traditional Chinese medicine, Dendrobium huoshanense (DHS) modulates intestinal flora, maintains the intestinal mucosal [...] Read more.

Background/Objectives: Sleep deprivation (SD) induces the accumulation of reactive oxygen species (ROS) in the intestine, causing inflammation in the intestine, thereby damaging the intestinal epithelial barrier function. As a traditional Chinese medicine, Dendrobium huoshanense (DHS) modulates intestinal flora, maintains the intestinal mucosal barrier, and promotes gastrointestinal motility and digestive secretion. However, the role and mechanism of DHS in improving SD-induced intestinal injury have not been fully studied. Methods: The SD model was established by subjecting rats to complete SD using a specialised SD instrument. Hematoxylin and eosin (HE) staining was performed to evaluate pathological injury in ileal tissues. Enzyme-linked immunosorbent assay (ELISA) and biochemical methods were used to quantify the main inflammatory cytokines, oxidative stress markers, and hypothalamic–pituitary–adrenal (HPA) axis activity. The expression levels of E-cadherin and Occludin proteins in the ileum tissue were analyzed by Western blotting. Additionally, the pH value of ileal mucus, unit secretion, water content, and dry matter weight were measured. Differential metabolites in rat ileum mucus were profiled using ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Results: DHS alleviated the pathological injury of the ileum induced by SD. DHS reduced the levels of serotonin (5-HT), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), while increasing interleukin-10 (IL-10) levels, thereby attenuating systemic inflammatory responses. Furthermore, DHS decreased malondialdehyde (MDA) content and elevated glutathione (GSH) and superoxide dismutase (SOD) levels in ileal tissues. DHS also upregulated the protein expression of E-cadherin and Occludin in intestinal tissues. In addition, DHS decreased the pH of ileal mucus, promoted intestinal mucus secretion, and increased dry matter content, facilitating the restoration of the mucus barrier. DHS may alleviate SD-induced ileal injury by modulating steroid hormone biosynthesis. DHS decreased the levels of adrenocorticotropic hormone (ACTH), cortisol (CORT), and corticotropin-releasing hormone (CRH), indicating that DHS suppresses the abnormal activation of the hypothalamic–pituitary–adrenal (HPA) axis. Conclusions: In this study, a comprehensive multi-index evaluation showed that DHS could significantly improve the ileal injury caused by SD in rats. The mechanism involved regulating the balance of serum neurotransmitters and inflammatory factors, reducing oxidative stress in tissues, and improving the physicochemical properties of intestinal mucus. Metabolomic analysis further revealed that these protective effects may be mediated via the regulation of steroid hormone biosynthesis pathways and are associated with the inhibition of abnormal HPA axis activation. Full article

(This article belongs to the Section Pharmacology and Drug Metabolism)

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20 pages, 3625 KB

Open AccessArticle

Transcriptome and Metabolome Analyses Reveal the Molecular Relationship Between Dietary Crude Protein Level and Liver Metabolism in Fattening Hu Sheep

by Patiguli Abudukeyimu, Fengmei Xie, Yifan Hu, Haiying He, Cheng Hou, Yiming Sulaiman, Huiguo Yang and Gao Gong

Metabolites 2026, 16(6), 375; https://doi.org/10.3390/metabo16060375 - 29 May 2026

Abstract

Background: Dietary crude protein (CP) acts as a key nutritional factor that affects the growth performance and liver metabolism of fattening Hu sheep, with metabolizable energy (ME) representing a major confounding factor in CP-related responses. To isolate the specific effects of CP on [...] Read more.

Background: Dietary crude protein (CP) acts as a key nutritional factor that affects the growth performance and liver metabolism of fattening Hu sheep, with metabolizable energy (ME) representing a major confounding factor in CP-related responses. To isolate the specific effects of CP on liver metabolism and minimize energy–protein interactions, we standardized dietary ME at 9.4 MJ/kg dry matter. Methods: We then established three isoenergetic CP concentrations: 11.07%, 13.07%, and 15.11%. A total of ninety 4-month-old male Hu sheep (with an initial body weight of 27.09 ± 1.83 kg) were allocated at random to three dietary treatment groups, each containing 30 animals distributed across three replicate pens, and fed pelleted total mixed rations (PTMRs) for 75 days under pen conditions in southern Xinjiang. Exploratory combined transcriptomic and metabolomic profiling of liver tissue was conducted to characterize how graded CP levels modulate growth traits and hepatic metabolic pathways, thereby identifying the appropriate dietary CP level for efficient and sustainable fattening of Hu sheep in this region. Result: Results indicated that animals fed the 15.11% CP diet showed a significantly higher average daily gain (ADG) and cumulative weight gain compared with those fed 11.07% or 13.07% CP (p < 0.05). Exploratory multi-omics enrichment analysis demonstrated significant overrepresentation (p < 0.05) of differentially expressed genes and metabolites in key biological pathways—including bile secretion, AMP-activated protein kinase (AMPK) signaling, steroid biosynthesis, peroxisome proliferator-activated receptor (PPAR) signaling, and oxidative stress-related and oxidative phosphorylation. Correlation analyses characterized two hub genes—ATP6AP1 and LOC101119853—that were significantly and negatively correlated with ADG (p < 0.05), whereas two metabolites—calcidiol and ADP—displayed significant positive relationships with ADG (p < 0.05). Pathway-level comparisons further demonstrated that both the 13.07% vs. 15.11% CP and the 11.07% vs. 15.11% CP contrasts yielded significant enrichment in AMPK signaling and steroid biosynthesis. Notably, calcidiol and ADP both declined numerically in the 13.07% vs. 15.11% CP comparison, whereas only ADP reached statistical significance in the 11.07% vs. 15.11% CP contrast. Conclusions: Collectively, under an ME level of 9.4 MJ/kg, a dietary CP concentration of 15.11% contributes to favorable growth of 4-month-old fattening Hu sheep housed in pens in southern Xinjiang. This level is associated with improved growth performance and coordinated regulation of central hepatic regulatory networks—particularly those involved in energy homeostasis and steroidogenesis—thereby supporting metabolic stability without compromising animal health or production efficiency. These findings provide a preliminary molecular basis for precision protein nutrition in Hu sheep feeding systems and offer translational insights for optimizing ruminant nutrition under arid and semi-arid environmental constraints. All correlations indicate potential associations, not causal relationships. Full article

(This article belongs to the Special Issue Metabolic Responses to Feed and Nutrition in Livestock)

17 pages, 4034 KB

Open AccessArticle

Transcriptomic and Metabolomic Insights into the Enhanced Quality of Anoectochilus roxburghii Seedlings in Sugar-Free Versus Conventional Tissue Culture Systems

by Xiangtao Chen, Fangfang Chen, Chuanzhi Kang, Tongwei Lin, Hongyang Wang, Yiheng Wang, Dehua Wu, Wanying Duan, Zekun Zhang and Chengcai Zhang

Metabolites 2026, 16(6), 374; https://doi.org/10.3390/metabo16060374 - 29 May 2026

Abstract

Background/Objective: Anoectochilus roxburghii, a high-value medicinal orchid, faces significant challenges in quality standardization during large-scale tissue culture due to a lack of understanding of the underlying molecular mechanisms. This study aimed to compare “Jianlan No.2” plantlets cultured under a conventional tissue [...] Read more.

Background/Objective: Anoectochilus roxburghii, a high-value medicinal orchid, faces significant challenges in quality standardization during large-scale tissue culture due to a lack of understanding of the underlying molecular mechanisms. This study aimed to compare “Jianlan No.2” plantlets cultured under a conventional tissue culture system (CK) and a sugar-free tissue culture system (TD), to elucidate the phenotypic and molecular basis for quality improvement. Methods: A systematic comparison was conducted. Phenotypic traits of plantlets from both systems were measured. Integrated transcriptomic (RNA sequencing) and untargeted metabolomic analyses were employed to identify the molecular differences at the gene expression and metabolite accumulation levels. Results: TD-grown seedlings exhibited significantly superior growth characteristics, including greater plant height, higher rooting rate, and improved transplant survival. Transcriptomic analysis identified 416 differentially expressed genes (DEGs) (44 upregulated, 372 downregulated in TD), which were significantly enriched in pathways related to cell wall organization, apoplast, and photosynthesis. Sixteen key genes were pinpointed as closely associated with seedling growth and metabolic regulation. Metabolomic profiling revealed 502 differentially accumulated metabolites (DAMs), with significant perturbations primarily in phenylpropanoid biosynthesis and terpenoid metabolism. Conclusions: The sugar-free tissue culture system enhances A. roxburghii seedling quality by coordinately modulating photosynthetic capacity, carbon metabolism, and the biosynthesis of key secondary metabolites. These findings provide a crucial molecular foundation for optimizing tissue culture protocols and advancing the standardized, high-quality cultivation of this valuable medicinal plant. Full article

(This article belongs to the Section Plant Metabolism)

11 pages, 1034 KB

Open AccessArticle

Humanin Restores Metabolic Hormone Homeostasis of Leptin, Ghrelin, Irisin and Asprosin in Streptozotocin-Induced Diabetic Mice

by Ferah Bulut, Muhammed Adam, Aslısah Ozgen and Mete Ozcan

Metabolites 2026, 16(6), 373; https://doi.org/10.3390/metabo16060373 - 29 May 2026

Abstract

Objective: Diabetes mellitus is closely associated with mitochondrial dysfunction, which disrupts cellular energy metabolism and perturbs hormonal homeostasis. Humanin (HN), a 24-amino acid peptide encoded within the mitochondrial genome, has attracted considerable attention due to its cytoprotective and metabolic regulatory properties. Despite [...] Read more.

Objective: Diabetes mellitus is closely associated with mitochondrial dysfunction, which disrupts cellular energy metabolism and perturbs hormonal homeostasis. Humanin (HN), a 24-amino acid peptide encoded within the mitochondrial genome, has attracted considerable attention due to its cytoprotective and metabolic regulatory properties. Despite its recognized biological potential, the role of HN in coordinating key metabolic hormone networks under diabetic conditions remains poorly understood. This study aimed to investigate the integrative effects of repeated humanin administration on key metabolic hormones leptin, ghrelin, irisin, and asprosin and its potential role in restoring hormonal homeostasis in a streptozotocin (STZ)-induced diabetic mouse model. Materials and Methods: Forty male mice were randomly assigned to four groups (n = 10 for each group): control, HN (4 mg/kg/day), STZ (150 mg/kg), and STZ + HN. Humanin was administered intraperitoneally for 15 consecutive days. Serum levels of leptin, asprosin, irisin, and ghrelin were measured using enzyme-linked immunosorbent assay (ELISA), and data were analyzed using one-way ANOVA followed by Tukey’s post hoc test. Results: STZ-induced diabetes markedly disrupted metabolic hormone balance, as indicated by decreased leptin and irisin levels and increased asprosin concentrations. Repeated HN treatment effectively restored leptin levels and suppressed asprosin concentrations, while irisin levels showed a relative increase compared to the STZ animals. In addition, ghrelin levels were significantly elevated in HN-treated diabetic mice compared to untreated STZ animals. Conclusions: These findings indicate that humanin exerts an integrative, multi-hormonal regulatory effect, supporting the restoration of metabolic and endocrine homeostasis under diabetic conditions. Full article

(This article belongs to the Section Pharmacology and Drug Metabolism)

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13 pages, 1189 KB

Open AccessArticle

Screening and Stratification Utility of GDF-15 and FGF-21 in Individuals Evaluated for Suspected Mitochondrial Disease: A Malaysian Cohort Study

by Affandi Omar, Dyg Pertiwi Abg Kamaludin, Wan Ahmad Syazani Mohamed, Fatimah Diana Amin Nordin, Rosnani Mohamed, Badrul Hisyam Razali, Imilia Ismail, Ngu Lock Hock and Julaina Abdul Jalil

Metabolites 2026, 16(6), 372; https://doi.org/10.3390/metabo16060372 - 29 May 2026

Abstract

Background/Objectives: Early detection of mitochondrial disorders remains challenging due to phenotypic heterogeneity and limited access to definitive molecular diagnostics. Circulating biomarkers such as growth differentiation factor-15 (GDF-15) and fibroblast growth factor-21 (FGF-21) have emerged as potential adjunct indicators. This study evaluated the [...] Read more.

Background/Objectives: Early detection of mitochondrial disorders remains challenging due to phenotypic heterogeneity and limited access to definitive molecular diagnostics. Circulating biomarkers such as growth differentiation factor-15 (GDF-15) and fibroblast growth factor-21 (FGF-21) have emerged as potential adjunct indicators. This study evaluated the screening and stratification utility of GDF-15 and FGF-21 in individuals assessed for suspected mitochondrial disease. Methods: Archived biological specimens collected between 2016 and 2017 were analysed from 221 individuals stratified into clinically high-risk, screen-positive non-high-risk, post-mortem unexplained death and healthy controls groups. Plasma and fibroblast lysate concentrations of GDF-15 and FGF-21 were quantified using enzyme-linked immunosorbent assays. Biomarker performance was assessed using receiver operating characteristic (ROC) analysis, comparative group analysis and correlation testing across clinically defined referral groups. Results: Both biomarkers were significantly elevated in clinically high-risk and screen-positive individuals compared with controls. GDF-15 demonstrated better discriminatory performance than FGF-21, with an area under the curve (AUC) of 0.7187 ± 0.0556 versus 0.6301 ± 0.0603. At a threshold of 300 pg/mL, GDF-15 demonstrated high sensitivity with moderate specificity for differentiation between clinically defined high-risk individuals and controls. Correlation analysis showed weak associations between GDF-15 and lactate and ammonia, while FGF-21 correlated modestly with glucose and alkaline phosphatase. A moderate positive correlation was observed between GDF-15 and FGF-21 across the overall cohort. Conclusions: GDF-15 and, to a lesser extent, FGF-21 may support early screening and stratification of individuals evaluated for suspected mitochondrial disease and assist in prioritising cases for further diagnostic evaluation. Full article

(This article belongs to the Special Issue The Future Perspective on Screening and Diagnosis of Inborn Errors of Metabolism)

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12 pages, 2940 KB

Open AccessSystematic Review

Probiotics After Metabolic and Bariatric Surgery: A Systematic Review and Meta-Analysis of Randomized Controlled Trials

by Mohammed Y. Ezzi

Metabolites 2026, 16(6), 371; https://doi.org/10.3390/metabo16060371 - 29 May 2026

Abstract

Background/Objectives: Patients undergoing metabolic and bariatric surgery (MBS) are at risk of micronutrient deficiencies and gut dysbiosis. Probiotics (such as Lactobacillus, Bifidobacterium) have been proposed as adjunct therapy to optimize postoperative outcomes. This review aimed to evaluate the effect of postoperative probiotic supplementation [...] Read more.

Background/Objectives: Patients undergoing metabolic and bariatric surgery (MBS) are at risk of micronutrient deficiencies and gut dysbiosis. Probiotics (such as Lactobacillus, Bifidobacterium) have been proposed as adjunct therapy to optimize postoperative outcomes. This review aimed to evaluate the effect of postoperative probiotic supplementation on anthropometric, metabolic, inflammatory, and micronutrient outcomes in MBS patients. Methods: Nine electronic databases were systematically searched, including PubMed, Web of Science, Cochrane Library, Google Scholar, Popline, Global Health Library, Virtual Health Library, New York Academy of Medicine, and OpenGrey, from inception through October 2024. Only randomized controlled trials (RCTs) were included. The Cochrane Collaboration risk-off-bias tool was used for quality assessment. Meta-analyses were performed using Comprehensive Meta-Analysis software version 2. Fixed-effects or random-effects models based on heterogeneity (I² threshold: 50%) were applied. Mean differences (MD) and 95% confidence intervals (CI) were calculated for all continuous variables. Results: Thirteen RCTs encompassing 666 patients (probiotics group: n = 344; control group: n = 322) were included. Incomplete outcome data represented the most prevalent high-risk domain (23%). Probiotic supplementation was associated with significantly improved serum vitamin D (MD: 25.32 nmol/L, 95% CI: 6.96–43.67, p = 0.007) and vitamin B12 levels (MD: 39.36 pg/mL, 95% CI: 1.88–76.84, p = 0.04). No statistically significant differences were observed in anthropometric outcomes (%EWL, BMI, weight, or waist circumference), lipid profile, glycemic indices, or inflammatory markers (TNF-α, IL-6, CRP). Conclusions: Postoperative probiotic supplementation may significantly improve vitamin D and B12 levels in patients undergoing MBS, suggesting a supportive role in mitigating micronutrient deficiencies. However, these findings should be interpreted with caution due to substantial heterogeneity across studies. Probiotics did not significantly affect weight loss, metabolic parameters, or inflammatory markers. Clinicians may consider probiotics as an adjunct strategy to support micronutrient status in at-risk postoperative patients. Large-scale, strain-specific trials incorporating standardized dietary control and microbiome profiling are warranted. Full article

(This article belongs to the Special Issue Metabolite Profiles in Inflammatory Diseases)

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27 pages, 2091 KB

Open AccessArticle

Acute Capillary Plasma Biomarker, Neuromuscular, and Perceptual Responses to Standardised Soccer Match Play in Elite Players: A Descriptive Study of Asynchronous Multi-Domain Recovery

by Lun Du, Jie Xiao, Chunpeng Li, Shuning Liu, Yaji Jiang, Yue Dou, Haotian Zhao, Wen Zhong, Kai Zhao and Chang Liu

Metabolites 2026, 16(6), 370; https://doi.org/10.3390/metabo16060370 - 29 May 2026

Abstract

Background: Soccer match play induces substantial mechanical, metabolic, inflammatory, and neuromuscular stress, yet post-match monitoring in applied settings often relies on isolated markers, venous sampling, or limited time points. This observational repeated-measures study aimed to describe whether capillary-derived biomarkers, neuromuscular performance, and perceptual [...] Read more.

Background: Soccer match play induces substantial mechanical, metabolic, inflammatory, and neuromuscular stress, yet post-match monitoring in applied settings often relies on isolated markers, venous sampling, or limited time points. This observational repeated-measures study aimed to describe whether capillary-derived biomarkers, neuromuscular performance, and perceptual measures showed asynchronous recovery during the first 48 h after a standardised soccer match in elite players. Methods: Twenty-two elite male outfield soccer players completed a standardised 90 min match. Capillary blood biomarkers, countermovement jump (CMJ), 20 m sprint performance, maximal voluntary contraction (MVC), and delayed onset muscle soreness (DOMS) were assessed before the match, immediately post-match, and at 24 and 48 h post-match. Time effects were analysed using repeated-measures mixed-effects models, and associations between biochemical and functional responses were examined descriptively. Results: Match play induced clear but domain-specific disturbances. IL-6 and cortisol rose rapidly immediately post-match, whereas hsCRP, CK, LDH, myoglobin, and DOMS showed delayed peaks during early recovery. CK, LDH, myoglobin, and soreness remained above baseline at 48 h. CMJ and sprint performance were impaired after the match but largely recovered by 48 h, whereas MVC showed its greatest decrement at 24 h. Exploratory associations indicated that larger muscle damage responses tended to co-occur with greater strength and jump decrements and higher soreness, but these analyses were not causal. Conclusions: Recovery after a standardised elite soccer match was multidimensional and non-synchronous across physiological, neuromuscular, and perceptual domains. A capillary-based, multi-domain assessment strategy may provide a feasible descriptive perspective for field-based observation of post-match fatigue. Full article

(This article belongs to the Special Issue Metabolic Adaptations to Exercise: Mechanisms, Modulators, and Health Impacts)

14 pages, 1192 KB

Open AccessArticle

Metabolomics Analysis of Aged Garlic Extract for the Identification of Novel Compounds

by Masato Nakamoto, Tsubasa Nishimura, Masahiro Ohtani and Toshiaki Matsutomo

Metabolites 2026, 16(6), 369; https://doi.org/10.3390/metabo16060369 - 29 May 2026

Abstract

Background/Objectives: Aged garlic extract (AGE), produced by aging raw garlic in an aqueous ethanol solution for over 10 months, exhibits multiple pharmacological activities, including antioxidant and anti-inflammatory effects. However, because AGE has a complex composition and many constituents remain insufficiently characterized, the chemical [...] Read more.

Background/Objectives: Aged garlic extract (AGE), produced by aging raw garlic in an aqueous ethanol solution for over 10 months, exhibits multiple pharmacological activities, including antioxidant and anti-inflammatory effects. However, because AGE has a complex composition and many constituents remain insufficiently characterized, the chemical basis underlying its broad activities is not fully understood. This study aimed to investigate these previously overlooked compounds in AGE to better understand its chemical complexity. Methods: AGE was fractionated using bioactivity assays to select target fractions for detailed chemical analysis. Metabolomics profiling was performed using liquid chromatography-mass spectrometry (LC-MS). Compounds were tentatively identified through database matching, fragmentation pattern analysis, and comparison with authentic standards. Results: Thirteen compounds not previously reported in AGE were tentatively identified. Citric acid was present at high levels. Citrulline and galacturonic acid were detected in AGE but not in raw garlic, suggesting that they are formed during the aging process. Trigonelline was detected and tentatively identified in the AGE sample used in this study. The remaining compounds included choline, 5-oxoproline, malic acid, gluconic acid, adenine, succinic acid, mucic acid, pipecolinic acid, and caffeic acid. These compounds may contribute to the diverse biological activities of AGE. Conclusions: These findings expand the chemical characterization of AGE and provide a foundation for understanding its broad pharmacological activities. They may also support future studies on functional food development and the health benefits of AGE. Full article

(This article belongs to the Section Food Metabolomics)

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19 pages, 1048 KB

Open AccessArticle

Integrating Sensory Evaluation and Metabolomics to Reveal the Metabolic Basis of Taste and Flesh Color in Melon (Cucumis melo L.)

by Yu Zhou, Binbin Li, Weizhong He, Fengjuan Liu, Yingying Fan, Jiangtao Du, Xing Cui, Weijia Lian, Qi Shen, Yan Wang, Zhongkai Zhao and Cheng Wang

Metabolites 2026, 16(6), 368; https://doi.org/10.3390/metabo16060368 - 28 May 2026

Abstract

Background: The sensory quality of melon (Cucumis melo L.) is determined by the complex interplay of metabolites within the fruit. However, the underlying metabolic mechanisms based on consumer sensory experience remain underexplored. Methods: Sensory evaluation was conducted on twelve melon [...] Read more.

Background: The sensory quality of melon (Cucumis melo L.) is determined by the complex interplay of metabolites within the fruit. However, the underlying metabolic mechanisms based on consumer sensory experience remain underexplored. Methods: Sensory evaluation was conducted on twelve melon cultivars, recording flesh color and quantitatively scoring acidity, sweetness, firmness, and aroma intensity. Based on the sensory results, eight cultivars were selected to establish two contrasting groups: sweet-type vs. acidic-type and orange-fleshed vs. green-fleshed. Untargeted metabolomics (UPLC-QTOF-MS) was then performed to analyze the samples, and differential metabolites were screened using OPLS-DA combined with univariate analysis. Results: Pathway enrichment analysis revealed that the key distinction between sweet and acidic taste profiles was associated with the specific accumulation of citric acid within the tricarboxylic acid (TCA) cycle in the acidic-type group. Regarding flesh color, the orange-fleshed group was enriched with carotenoid derivatives like β-citraurinene and the oxidized tocopherol product α-tocopherolquinone, whereas the green-fleshed group mainly accumulated phytol, a chlorophyll degradation product, along with more abundant terpenoids. Conclusions: By integrating sensory phenotyping with metabolomic analysis, this study identified key differential metabolites and candidate pathways associated with taste and color in melon, providing metabolic insights and data resources for quality evaluation and regulation. Full article

(This article belongs to the Section Food Metabolomics)

15 pages, 2386 KB

Open AccessArticle

Enhanced Synthesis of Polyphenols and Terpenes by UV-A Irradiation in Artemisia argyi Leaves

by Shaozheng Li, Zikun Zhang, Lanqi Yang, Heyang Wang, Haike Gu and Junfeng Liu

Metabolites 2026, 16(6), 367; https://doi.org/10.3390/metabo16060367 - 28 May 2026

Abstract

Background: Secondary metabolites not only constitute the material basis for plant responses to multiple environmental stresses but are also extensively utilized in the pharmaceutical industry. Methods: In the present work, we investigated the metabolic response of Artemisia argyi to UV-A irradiation through transcriptomic [...] Read more.

Background: Secondary metabolites not only constitute the material basis for plant responses to multiple environmental stresses but are also extensively utilized in the pharmaceutical industry. Methods: In the present work, we investigated the metabolic response of Artemisia argyi to UV-A irradiation through transcriptomic and metabolomic analyses. Results: After 16 h of UV-A treatment with an intensity of 2.5 μmol m⁻² s⁻¹ and 8 h of dark cultivation, a total of 4343 differentially expressed genes were identified, most of which were associated with fatty acid metabolism, biosynthesis of secondary metabolites, and ribosome. Of the 1959 metabolites detected in samples exposed to a 16/8 h UV-A/dark cycle for 6 days, a total of 223 differentially accumulated metabolites were identified and classified into 12 subgroups, with phenolic acids and flavonoids representing the largest subgroups. Comprehensive analyses indicated that polyphenols and terpenes play critical roles in the adaptation of A. argyi to UV-A irradiation. The phytohormone methyl jasmonate was identified as a key regulator of the enhanced synthesis of these secondary metabolites, through activation of transcription factors from the MYB and bHLH families. Conclusions: This study deepens our understanding of secondary metabolic regulation in response to UV-A stress and provides a simple and reliable method to promote the accumulation of specific secondary metabolites in Artemisia species. Full article

(This article belongs to the Section Plant Metabolism)

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