Search Results (293)

Oxidative stress plays an important role in systemic lupus erythematosus (SLE). To elucidate whether it is already present in early phases, we investigate oxidative stress-related genes in incomplete SLE (iSLE) and quiescent SLE (qSLE, inactive disease), as well as the effect of nuclear factor erythroid-derived 2-like 2 (NRF2) activators on NRF2-related genes in peripheral blood mononuclear cells (PBMCs) and HaCaT keratinocytes. In total, 28 qSLE patients, 29 iSLE patients and 21 age- and sex-matched healthy controls (HCs) were included. Serum free thiols, reactive oxygen species (ROS) levels and NRF2-related antioxidant gene expression were measured. Furthermore, PBMCs and HaCaT keratinocytes were treated with the NRF2 activators sulforaphane (SFN) and dimethyl fumarate (DMF) in vitro to assess expression of antioxidant genes. Finally, NRF2 and Heme oxygenase-1 (HMOX1) proteins in non-sun-exposed skin sections were assessed. Thiols were significantly lower in qSLE patients compared to HCs. In whole blood, Kelch-like ECH-associating protein 1 (KEAP1) and catalase (CAT) mRNA levels were significantly reduced in iSLE and qSLE. Treatment with SFN or DMF in PBMCs upregulated HMOX1 and NAD(P)H quinone dehydrogenase-1 (NQO1) mRNA expression and downregulated CAT expression. In HaCaT cells, mRNA expression of HMOX1, NQO1 and thioredoxin was upregulated. There were no differences in protein expression of NRF2 and HMOX1 in skin tissues. In conclusion, in qSLE patients, oxidative stress is elevated, while antioxidant capacity is decreased. A similar trend, although not significant, is seen in iSLE patients, which indicates that redox imbalances are already present in early phases, but not as obvious as in established SLE. NRF2 activators upregulate antioxidant gene expression in PBMCs and HaCaT cells, highlighting their potential role to modulate oxidative stress pathways in SLE. Full article

Heme oxygenase-1 (HO-1), encoded by HMOX1, catalyzes the rate-limiting step of heme degradation and generates biliverdin, carbon monoxide, and ferrous iron, thereby linking heme turnover with redox regulation and stress-associated signaling. In birds, biliverdin is retained as a major heme-derived product, but the cellular consequences of HMOX1 perturbation remain insufficiently defined. Here, CRISPR/Cas9-mediated editing was used to generate a heterogeneous HMOX1-edited population in Chicken hepatocellular carcinoma-derived cells. The selected sgRNA reduced HO-1 protein abundance by approximately 47%, and no detectable cleavage was observed at the seven predicted high-risk off-target loci examined. Compared with vector-control cells, HMOX1-edited cells exhibited intracellular heme accumulation, reduced biliverdin levels, increased oxidation-sensitive fluorescence, and reduced CCK-8 absorbance values, indicating disruption of heme–biliverdin metabolic and redox homeostasis. RNA sequencing identified 2650 differentially expressed genes, including 951 upregulated and 1699 downregulated genes. Downregulated genes were mainly enriched in immune, cytokine, MAPK/stress, and extracellular signaling-associated pathways, whereas DNA replication and cell-cycle-related genes were increased. Enrichment-term association and STRING functional-association analyses further identified a coordinated module involving IL1B, JUN, NFKBIA, IRF1, TGFB1, IL10, CCL5, and PTGS2. Independent RT-qPCR analysis confirmed selected expression trends. These findings show that heterogeneous HMOX1 editing and reduced HO-1 abundance are associated with disruption of the avian heme–biliverdin metabolic axis and coordinated remodeling of basal immune, stress, extracellular signaling, and cell-cycle-associated transcriptional programs in Chicken hepatocellular carcinoma-derived cells. Full article

This study evaluated dietary quercetin supplementation to investigate the effects of increasing inclusion levels on growth performance, diarrhea, serum health-related indicators, and whole-blood gene expression in weaned piglets. In a 28-day feeding trial, 128 piglets received diets formulated to contain 0, 100, 200, or 300 mg/kg quercetin (four pens of eight piglets per treatment; pen as the experimental unit). Final body weight and average daily gain were higher in the 200 and 300 mg/kg groups than in the control, and the 300 mg/kg group had the highest gain-to-feed ratio (G:F) (p < 0.05). Diarrhea rate declined linearly (p < 0.001); the rate in the 300 mg/kg group was lower than in the control and 100 mg/kg groups but did not differ from the 200 mg/kg group. Quercetin reduced serum malondialdehyde, pro-inflammatory cytokines, urea nitrogen, and aspartate aminotransferase, while increasing total protein, albumin, total antioxidant capacity, several antioxidant enzyme activities, and interleukin-10 (p < 0.05). Serum diamine oxidase activity and D-lactate concentration also declined linearly (p < 0.001). Whole-blood expression of PTGS2 decreased; HMOX1, NFE2L2, and NQO1 increased; and SOD1 was unchanged. Overall, within the tested range, increasing dietary quercetin improved growth and feed efficiency, reduced diarrhea, and was accompanied by coordinated changes in serum health-related indicators and whole-blood transcripts. Full article

Identifying physiological changes during the transition period is essential for improving the health and productivity of dairy goats. This study evaluated hematological, biochemical, hormonal, oxidative stress, and molecular alterations in Shami goats during the pre-pregnancy, late pregnancy, and early lactation periods. Eighty clinically healthy goats were examined, and blood samples were analyzed for hematological indices, metabolic and hormonal profiles, oxidative stress biomarkers, and relative expression of genes associated with energy metabolism, antioxidant defense, inflammation, and autophagy. Late pregnancy was characterized by significant (p < 0.05) increases in red blood cell count (RBCs), hemoglobin concentration (Hb), neutrophils, albumin, globulin, urea, insulin-like growth factor-1 (IGF-I), and malondialdehyde (MDA), accompanied by decreased glucose, cholesterol, total protein (TP), antioxidant markers, total leukocyte count, packed cell volume, and monocytes. Early lactation was associated with higher non-esterified fatty acid, triiodothyronine (T3), and thyroxine (T4) levels. Genes involved in lipid mobilization and oxidation, ketogenesis, inflammation, cellular stress, and autophagy; sirtuin 1 (SIRT1), peroxisome proliferator-activated receptor alpha (PPARA), carnitine palmitoyltransferase 1a (CPT1A), 3-hydroxy-3-methylglutaryl-coenzyme a synthase 2 (HMGCS2), cluster of differentiation 36 (CD36), lipase E (LIPE), protein kinase amp-activated catalytic subunit alpha 1 (PRKAA1), solute carrier family 2 member 1 (SLC2A1), haptoglobin (HP), interleukin 6 (IL6), heat shock protein 70 (HSP70), heme oxygenase 1 (HMOX1), beclin 1 (BECN1), and autophagy-related protein 5 (ATG5) were significantly upregulated, whereas antioxidant- and glucose transport-related genes nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione peroxidase 1 (GPX1), catalase (CAT), thioredoxin (TXN), and solute carrier family 2 member 4 (SLC2A4) were downregulated during the transition period. The results indicate well-orchestrated metabolic and molecular adaptations that can be employed as biological indicators to track the physiological status of Shami goats. These findings fulfilled the study objective and identified potential biomarkers of the transition period in Shami goats. Full article

The increased activity of δ-aminolevulinic acid synthase-1 (ALAS1) leads to the accumulation of δ-aminolevulinic acid (ALA), which may reduce the iron(III) retention ability of ferritin, resulting in iron(III) overload. The accumulation of ALA occurs in the condition of attenuation of the heme negative feedback loop over ALAS1 activity in concert with the induction of ALAS1 expression. Attenuation of the heme negative feedback loop is maintained by elevated quantities of the heme catabolizing enzyme, heme oxygenase-1 (HO-1), which, in turn, may be induced by highly increased heme concentration. The upregulation of brain HO-1 occurs in patients with Alzheimer’s and Parkinson’s diseases. A mouse with overexpressed human HMOX1 is a model of schizophrenia with concurrent iron overload. HO-1 inhibitors reduce oxidative damage to whole cells and mitochondrial compartments of rat astrocytes transfected with the HMOX1 gene. Iron reduction within the heme prosthetic moiety of P450 cytochromes in microsomes is facilitated by NADPH-cytochrome P450 oxidoreductase (CPR), while adrenodoxin reductase performs this function in the mitochondria. In partial CPR-deficient conditions, the half-life of apocytochromes is prolonged while HO-1 is induced due to the elevated heme release from unreduced cytochromes. This study posits that, before being degraded by HO-1, the released hemin triggers the oligomerization of cytochromes, as well as other oxidative damages, by producing hydroperoxyl radicals from hydrogen peroxide produced in uncoupling reaction. After reaching a specific level, iron(III) overload may trigger the saturation of CPR, resulting in the development of neuropathologies. Full article

Background/Objectives: This study aimed to identify common target genes, molecular interaction networks, and signaling pathways shared across the investigated toxic metal(loid)s—lead (Pb), cadmium (Cd), methylmercury compounds (MMC), arsenic (As), nickel (Ni), and hexavalent chromium (Cr(VI))—and associated with lung cancer overall, non-small-cell lung cancer (NSCLC), and small-cell lung cancer (SCLC). Methods: The Comparative Toxicogenomics Database (CTD) was used to identify associated genes, GeneMANIA to characterize molecular interaction networks, and ToppFun for pathway enrichment analysis. Results: Two common genes (MTOR and TP53) were identified for SCLC, eight for NSCLC (CAT, CCNB1, MTOR, NQO1, SOD2, STAT3, TP53, and VEGFA), and eighteen for lung cancer overall. Gene network analysis revealed predominantly physical interactions in SCLC, pathway interactions in NSCLC, and co-expression in lung cancer overall. Several toxic metal(loid)s exhibited concordant effects on TP53, VEGFA, HMOX1, and IL6 expression. TP53 was the only gene common to all investigated toxic metal(loid)s and lung cancer categories. Pathway enrichment identified PI3K–AKT–mTOR signaling in both SCLC and NSCLC, VEGF signaling in NSCLC, and MAPK, interleukin, and macrophage-stimulating protein (MSP) signaling in lung cancer overall. Conclusions: The identified genes and pathways suggest molecular mechanisms potentially relevant to the association between exposure to the investigated toxic metal(loid)s and lung carcinogenesis, particularly those related to cell survival, inflammation, oxidative stress, and angiogenesis. TP53 may represent a common molecular link warranting further investigation. The identified pathways represent candidate biomarkers and mechanistic targets, highlighting the biological heterogeneity of lung cancer and the importance of subtype-specific analyses. Full article

The vitamin D receptor (VDR) exerts pleiotropic effects essential for human health and modulates the transcription of nuclear factor erythroid 2-related factor 2 (Nrf2), a key regulator of the cellular antioxidant defense system. Polymorphisms represent a major source of interindividual variability and may influence susceptibility to oxidative stress. The VDR polymorphisms ApaI (A > C) and TaqI (T > C) have been associated with disease susceptibility; however, their role in oxidative stress responses during aging remains poorly understood. This study evaluated the association between ApaI and TaqI polymorphisms and susceptibility to oxidative stress-induced cell death, as well as the expression of genes within the VDR–Nrf2 signaling axis, in peripheral blood mononuclear cells (PBMCs) from cognitively healthy Chilean older adults. VDR genotyping was performed using real-time PCR (TaqMan SNP Genotyping Assay), PBMC viability following H₂O₂ exposure was assessed by flow cytometry, and gene expression levels were determined by quantitative PCR (qPCR). For ApaI, homozygous carriers of the A allele showed greater PBMC viability following H₂O₂ exposure (p < 0.0068) and higher mRNA expression levels of Nrf2 (p = 0.0176) and its downstream genes HMOX1 (p = 0.0070) and GST (p = 0.0047) compared with carriers of the C allele (AC or CC). For TaqI, homozygous carriers of the protective C allele exhibited greater PBMC viability (p < 0.0465) and higher Nrf2 (p = 0.0324), HMOX1 (p = 0.0206), and GST (p = 0.0029) expression compared with carriers of the risk T allele (TC or TT). These findings provide novel evidence that VDR polymorphisms may modulate antioxidant responses through regulation of Nrf2-dependent gene expression, suggesting a genetic determinant of oxidative stress resilience in cognitively healthy older adults. Full article

High-temperature stress poses a critical challenge to cold-water aquaculture; however, the size-associated molecular mechanisms underlying thermal adaptation in triploid rainbow trout (Oncorhynchus mykiss) remain poorly understood. Here, we integrated transcriptomic and metabolomic profiling of liver tissues, which plays central role in energy metabolism and stress integration, from three body-weight classes—small (0.8 kg ± 0.18 kg), medium (1.5 kg ± 0.22 kg), and large (2.5 kg ± 0.31 kg)—sampled under peak summer heat stress (20.5 °C). Transcriptomic analysis identified 974, 570, and 862 group-specific differentially expressed genes, respectively, revealing a non-linear, size-associated transcriptional pattern. Genes assigned to the ribosome pathway were significantly enriched in comparisons involving MLA and SLA relative to LLA, but not between SLA and MLA, whereas carbon metabolism and amino acid biosynthesis were enriched exclusively in smaller fish, indicating a higher catabolic burden under heat stress. Metabolomic profiling identified 1123 metabolites, with lipids accounting for 45.06%, and showed size-specific enrichments in biosynthesis of unsaturated fatty acids, glycerophospholipid metabolism, arachidonic acid metabolism, and pathways related to necroptosis. Integrative analysis revealed that in smaller fish, pla2g1b and gpx4a coordinately regulate the accumulation of prostaglandin H2 and 16(R)-HETE, forming a regulatory network with ferroptosis-related genes acsl4a and hmox1a; concurrently, chka, lpin1, and phospholipase A2 members drive extensive membrane phospholipid remodeling. The observed negative correlation between hsd17b3 expression and 7α-hydroxytestosterone levels suggests size-associated steroid-mediated energy repartitioning. Collectively, smaller fish undergo extensive transcriptional and metabolic reprogramming with heightened activation of cell death pathways, whereas larger fish maintain greater thermal buffering capacity. These findings provide molecular targets for size-stratified thermal management and selective breeding in rainbow trout aquaculture. Full article

Preeclampsia is a serious pregnancy complication characterised by maternal vascular dysfunction, placental dysfunction, and organ injury, with no effective treatment currently available. Kampo, a system of Japanese traditional medicine comprising standardised herbal formulations, could target pathophysiological pathways driving preeclampsia. We evaluated the effects of select Kampo formulations on markers of preeclampsia using primary human trophoblasts, placental explants, human umbilical vein endothelial cells (HUVECs), and uterine microvascular endothelial cells (UtMVECs). Twelve formulations were initially screened in HUVECs, and six formulations advanced for further study. TNFα was used to induce endothelial dysfunction, and angiogenic, antioxidant, inflammatory, and vascular dysfunction markers were assessed. Overall, Kampo formulations had minimal effect on sFlt-1 expression and only modest effects on sFlt-1 secretion by primary human trophoblast. In contrast, several formulations consistently increased placental growth factor (PlGF) expression and secretion, upregulated HMOX1 in trophoblasts, and enhanced PlGF secretion from placental explants. In endothelial cells, Kampo treatment partially reversed TNFα-induced dysfunction, demonstrated by reduced VCAM1 expression, and additional endothelial cell type-dependent effects on ET-1 and inflammatory pathways. These findings indicate that selected Kampo formulations modulate key pathways involved in the pathophysiology underpinning preeclampsia and warrant further investigation as potential therapeutic candidates. Full article

Background/Objectives: Chronic obstructive pulmonary disease (COPD) is increasingly viewed as a disorder of impaired cellular adaptation to chronic stress, involving oxidative injury, mitochondrial dysfunction, and accelerated cellular senescence. We investigated whether genetic variation in these pathways contributes to disease susceptibility, lung function impairment, and polygenic risk prediction. Methods: Thirty-three single-nucleotide variants were analysed in 747 patients with COPD and 703 controls. Associations with disease susceptibility and lung function parameters were assessed using regression models with correction for multiple testing. Weighted and unweighted polygenic scores were constructed from associated variants and evaluated using receiver operating characteristic and net reclassification improvement analyses. Results: Significant associations were identified in genes involved in antioxidant defence (NFE2L2, HMOX1, GSR), PI3K/AKT/mTOR signalling (PIK3R1, PTEN), mitochondrial function (TOMM40), cellular stress responses (FOXO3A), and long non-coding RNA regulation (MEG3, CDKN2B-AS1). The strongest association was observed for PIK3R1 rs831125 (OR = 2.31, p = 2.53 × 10⁻¹⁰). Variants in NFE2L2, PIK3R1, MEG3, MALAT1, and SIRT3 were additionally associated with pulmonary function parameters. The weighted polygenic score demonstrated good discriminative ability (AUC 68.8%, 95% CI 65.9–71.7%) and substantially improved prediction when combined with age, sex, and smoking exposure (AUC 88.1%, 95% CI 86.3–89.8%; NRI = 0.62, p = 2.21 × 10⁻²⁸). Conclusions: The identified loci converge on interconnected pathways involved in cellular stress adaptation, mitochondrial homeostasis, and senescence, supporting their contribution to chronic obstructive pulmonary disease susceptibility and functional decline. Full article

Background: Plasticizers, including phthalate esters and phthalate-free alternatives, are widely detected environmental chemicals. Although increasing evidence suggests that plasticizers may disrupt gastrointestinal homeostasis, their potential molecular links with inflammatory gastrointestinal disorders (IGDs) remain unclear. Methods: This study aimed to systematically identify potential molecular targets and pathways linking representative plasticizers with IGDs. An integrative network toxicology framework was applied to investigate four plasticizers, including dimethyl phthalate (DMP), diethyl phthalate (DEP), dioctyl phthalate/di(2-ethylhexyl) phthalate (DOP/DEHP), and acetyl tributyl citrate (ATBC), in relation to Crohn’s disease (CD), ulcerative colitis (UC), esophagitis, and gastritis. Plasticizer- and disease-related targets were collected from public databases, followed by overlapping target screening, protein–protein interaction network analysis, functional enrichment analysis, GEO-based transcriptomic validation, molecular docking, molecular dynamics simulation, and single-cell RNA-seq analysis. Results: Disease-specific candidate targets were identified, including CXCL8 and FN1 for CD, IL1B for UC, MAPK3, FASN, FN1, PPARG, CXCL8, FOS, and HIF1A for esophagitis, and MMP9, TNF, TLR4, IL6, CCR2, IFNG, and PTGS2 for gastritis. Cross-disease analysis further identified plasticizer-associated signature targets, including MMP7 for DMP, HMOX1 and NOS2 for DEP, and LTF and CCL11 for ATBC. Enrichment analysis indicated that these targets were mainly involved in inflammatory, chemokine, MAPK-related, and xenobiotic response pathways. Molecular docking and dynamics simulations suggested stable interactions between selected plasticizers and candidate targets, while single-cell analysis revealed their cell-type-specific expression patterns in epithelial, immune, and stromal compartments. Conclusions: This study provides an exploratory network toxicology framework for identifying potential molecular associations between plasticizer exposure and IGDs. The findings highlight disease-specific and plasticizer-associated candidate targets that may guide future experimental validation and environmental risk assessment. Full article

Androgenetic alopecia (AGA) is associated with follicular miniaturization, oxidative stress, and microinflammation. Hydroxytyrosol (HT) has antioxidant and anti-inflammatory activities, but its mechanisms in AGA remain unclear. Differentially expressed genes (DEGs) from GSE212301 were intersected with predicted HT targets to identify candidate targets. Enrichment analysis, PPI-based hub gene screening, molecular docking, and molecular dynamics (MD) simulations were performed. A total of 506 DEGs and 24 HT-related AGA candidate targets were identified. These targets were enriched in inflammation- and oxidative stress-related pathways, including MAPK, NF-κB, TNF, and HIF-1 signaling. HMOX1, PTGS2, FOS, and JUN were identified as hub genes. Docking showed favorable binding between HT and these targets, particularly PTGS2. The 50 ns MD simulation supported the relative stability of the HT–PTGS2 complex. HT may modulate AGA-related inflammatory and oxidative stress networks, particularly through HMOX1 and PTGS2, providing a basis for further experimental validation. Full article

Estimating the post-mortem interval (PMI) remains a major challenge in forensic pathology, particularly beyond the earliest postmortem phases. RNA-based markers measured by RT-qPCR have been proposed as potential tools for PMI estimation, but their reliability depends on technical robustness, reference gene stability, and data representation. Technical reproducibility was overall satisfactory. However, the candidate reference genes showed different temporal behaviors: ACTB remained relatively stable, whereas RPL4 displayed significant time-dependent drift, resulting in partial instability of the composite reference signal. All target genes were associated with PMI, with HPRT1 and HMOX1 emerging as the most informative markers. Several targets also showed evidence of non-linear temporal dynamics. In predictive analyses, models based on raw Ct values consistently outperformed ΔCt-based models. A parsimonious model based on HMOX1 and HPRT1 showed the most favorable trade-off between interpretability and predictive performance within this exploratory dataset, although prediction error remained non-negligible. These findings suggest that postmortem cardiac transcriptional profiles may contain temporal information useful for PMI-oriented modeling but also show that predictive performance remains limited by reference gene behavior, analytical strategy, and non-negligible estimation error. Nine porcine hearts were stored at 4 °C and sampled at 0, 12, 24, 48, 72, 96, and 120 h. RT-qPCR was performed in technical triplicate for selected target genes (BAX, CASP3, HIF1A, HMOX1) together with additional quantified transcripts (ACTB, RPL4, GAPDH, HPRT1). Technical reproducibility, reference gene stability, temporal trends and predictive performance were assessed using mixed-effects models and predictive models evaluated by leave-one-heart-out cross-validation. Comparative analyses were performed using raw Ct, ΔCt, and ΔΔCt data. Overall, postmortem cardiac RT-qPCR profiling should be regarded as a proof-of-concept framework developed under specific controlled refrigerated conditions. Therefore, further external validation under heterogeneous real-world forensic scenarios and methodological standardization are required before a real-life forensic application. Full article

Citrus flavonoid intake is associated with beneficial effects on endothelial function. Our previous randomized control trial demonstrated that the concentration of Hesperetin-7-O-glucuronide (H7G) was positively correlated with the improvement in endothelial function in overweight and obese participants following blood orange juice consumption. To explore the underlying mechanism by which H7G improves endothelial function, we investigated the regulation of H7G on endothelial function in a permanent human endothelial cell line (EA. hy926 cells) under normal and oxidative conditions treated with high-oxidation low-density lipoprotein. The results indicated that H7G improved the expression of nitric oxide synthase 3 (NOS3), heme oxygenase 1 (HMOX1) ad glutamate cysteine ligase catalytic (GCLC), and inhibited the expression of endothelin-1 (EDN1), through the upregulation of miR-660-5p and inhibition of miR-21-5p. In summary, H7G improves endothelial cell function via the upregulation of miR-660-5p and the inhibition of miR-21-5p. Full article

Heme metabolism in the liver has traditionally been described as a linear pathway that supports oxygen utilization, redox balance, and detoxification. Here, we synthesize recent evidence and propose a framework in which heme functions as a system-level regulator, the “queen” of metabolism, whereas its upstream intermediate protoporphyrin IX (PPIX) represents a chemically reactive “dark twin” that emerges when metabolic flux fails to resolve. In this view, metabolic state is defined not only by end products but also by the behavior of pathway intermediates. This system is spatially organized. Hepatocytes dominate heme synthesis and utilization. In contrast, liver stromal compartments, particularly Kupffer cells, play a central role in heme degradation through heme oxygenase-1 (HMOX1), linking heme turnover to iron recycling and stress adaptation. The metabolic state of the liver therefore reflects not only pathway flux but also the degree of coupling between these cellular compartments. We propose a state model of hepatic heme metabolism. In the resolution state, most evident during inflammation, coordinated hepatocyte–macrophage activity maintains flux and limits intermediate accumulation. In contrast, the expansion state, exemplified in cancer, is defined by impaired flux completion, leading to PPIX accumulation, metabolic heterogeneity, and oxidative stress. This framework reframes liver disease through intermediate behavior rather than pathway presence: porphyrias reflect direct overload, metabolic liver diseases partial expansion, and hepatocellular carcinoma a fully developed expansion state. By focusing on the “intermediate space,” this model links biochemistry, spatial organization, and disease pathogenesis, while suggesting new opportunities for diagnosis and therapy based on metabolic state. Full article