Search Results (103)

The application of natural bioactive compounds in cosmeceutical products, particularly as skin-lightening agents, has received increasing interest. Caulerpa racemosa, a green macroalga of the Chlorophyta division, contains beneficial nutrients that are applicable as food and cosmeceutical ingredients. This study investigated the in vitro effect of the ethanolic extract of C. racemosa (CR) on regulation of melanogenic-related signaling and gene expression in SK-MEL-5 human melanoma-derived cells. Identification of bioactive components revealed that catechin, rutin, and quercetin as flavonoid contents were found in CR extract, analyzed using HPLC. The expressions of microphthalmia-associated transcription factor (MITF), extracellular signal-regulated kinase (ERK) signaling molecules, and melanogenic-related molecules were analyzed via Western blotting and qPCR. The CR extract treatment applied to SK-MEL-5 cells decreased the MITF protein expression level, which correlated with increased pERK expression, and no cytotoxic effect was observed. The subsequent treatment reduced the expression of melanogenesis-related genes (TYR, TYRP1, MC1R, and DCT) that were downstream targets of MITF. This study provides preliminary evidence that CR extract may modulate melanogenesis-related signaling. However, the specific bioactive compounds responsible for the observed effects remain to be identified, as the extract contains a complex mixture of phytochemicals. Further fractionation studies are needed to pinpoint the active constituents. The variability of extract composition due to seasonal and geographical factors should be considered for future standardization. Full article

Excessive fructose consumption is associated with metabolic syndrome (MS). This study evaluated the effect of two coffee by-products, coffee pulp (CP) and coffee silverskin (SK), on fructose-induced intestinal changes. Sprague–Dawley rats were assigned to six groups (n = 6/group) for 10 weeks: Control, Fructose (FRU; 20% fructose in drinking water), CP, CP + FRU, SK, and SK + FRU. CP and SK were administered by oral gavage (250 mg/kg/day) using corn oil as vehicle. Intestinal morphology, gene expression (RT-qPCR), and gut microbiota composition (16S rRNA sequencing) were assessed. Fructose significantly increased jejunal expression of the glucose transporters SGLT1 and GLUT2. CP and SK reversed SGLT1 and GLUT2 overexpression and reduced GLUT5 expression relative to the FRU group. Fructose also markedly increased expression of sweet taste receptors TAS1R2 and TAS1R3 and the transcription factors SREBP-1c and ChREBP. Both CP and SK normalized TAS1R2 and TAS1R3 expression, whereas SK additionally prevented SREBP-1c and ChREBP overexpression. Both by-products restored fructose-induced reductions in microbial richness and alpha diversity. CP also modified beta diversity and increased the abundance of the genus Blautia compared with FRU. In conclusion, CP and SK reversed several fructose-induced intestinal alterations, namely in the jejunal expression of sugar-sensing and absorption-related genes. Additionally, CP showed microbiota-modulating effects, whereas SK modulated the jejunal expression of key transcription factors (SREBP-1c and ChREBP) involved in carbohydrate and lipid metabolism. Overall, these findings suggest that CP and SK may represent promising candidates for mitigating fructose-induced intestinal alterations. Full article

Antisense oligonucleotides (ASOs) are short, synthetic DNA fragments able to modulate gene expression. In this work, the cytosolic isoform of glucose-6-phosphate dehydrogenase (Cyt-G6PDH) was selected as a target for ASOs to induce transient gene silencing in barley (Hordeum vulgare). G6PDH is the most important enzyme of the oxidative pentose phosphate pathway (OPPP), supplying NADPH and regulating the entire cycle. Different ASOs were tested at different concentrations on the leaf surface. Their effects were evaluated by measuring enzymatic activity, gene expression, and protein abundance. Treatment with 30 µM ASOs for 6 h represented the optimal condition, inducing a 40–60% reduction in G6PDH total activity in barley leaves, with a specific decrease in the cytosolic isoform, as determined by DTT-sensitive enzymatic assays. ASOs designed on the main regulator of cyt-G6PDH, the shaggy-like kinase (SK11), led to an analogous effect in terms of G6PDH activity, confirming the involvement of HvSK11 in the regulation of cyt-G6PDH in barley. Consistently, qRT-PCR analyses showed that ASO treatment simulates an abiotic stress condition obtained by the down-regulation of Cyt-G6PDH. These results support the use of ASOs as a rapid and efficient method for the functional analysis of key metabolic regulators in plants to overcome complications in recalcitrant organisms or lethal genes. Full article

Paclitaxel (PTX), a taxane-derived chemotherapeutic agent, is frequently used in the treatment of breast cancer (BC). Its anticancer effects are primarily associated with microtubule stabilization, disruption of cell-cycle progression, and triggering of apoptotic cell death. In the present study, we investigated the effects of PTX on the expression of genes involved in cancer-related pathways, energy metabolism, and drug resistance in four molecularly distinct BC cell lines: MCF-7, BT-474, SK-BR-3, and MDA-MB-231. The half-maximal inhibitory concentrations (IC₅₀) of PTX in BC cell lines and the non-tumorigenic hTERT-HME1 breast epithelial cell line were determined by the MTT assay to assess cell cytotoxicity. BC cells were exposed to nine different concentrations of PTX for 24, 48, and 72 h to evaluate concentration- and time-dependent effects. Following treatment, total RNA was isolated and converted into cDNA, and RT-qPCR analysis was performed to investigate PTX-mediated alterations in the expression of genes associated with cancer-related pathways. The impact of PTX on the cell-cycle phase distribution and apoptotic cell death was evaluated by flow cytometry. Treatment with PTX for 48 h at concentrations of 12.60 nM in MCF-7, 5.09 nM in BT-474, 16.09 nM in SK-BR-3, and 36.66 nM in MDA-MB-231 cells reduced cell viability and increased apoptosis. PTX treatment also altered the expression of genes involved in apoptosis, cell-cycle regulation, angiogenesis, epithelial–mesenchymal transition, hypoxia-related signaling, energy metabolism, telomere maintenance, and therapy resistance. Collectively, these findings demonstrate that PTX elicits heterogeneous molecular and cellular responses across molecularly distinct BC cell lines, particularly in cell viability, apoptosis, metabolic regulation, and treatment response. These in vitro findings suggest potential molecular mechanisms that could explain why some cells are more sensitive to PTX than others, but further experimental and clinical validation is needed to confirm this. Full article

The study investigates the impact of molecular iodine (I₂) supplementation on the viability, invasiveness, and angiogenic potential of high-risk neuroblastoma (NB). In vitro assays were performed using NB cell lines SK-N-AS (non-MYCN-amplified) and SK-N-BE(2) (MYCN-amplified). The role of peroxisome proliferator-activated receptor gamma (PPARγ) was evaluated using the antagonist GW9662, gene expression (RT-qPCR), and protein levels (Western blot). In vivo, zebrafish xenografts were used to evaluate tumor size, angiogenesis, and caudal cell dissemination. I₂ supplementation significantly decreased cell viability in both cell lines, independent of PPARγ activation. In SK-N-BE(2), I₂ impaired cell migration, as measured by a wound-healing assay, in apparent independence of PPARγ activation. However, gene expression indicates that I₂ acts in complex ways, including direct antioxidant effects and PPARγ-mediated effects. The significant decrease in reactive oxygen species levels (DCFDA staining) and the silencing of the long noncoding RNA myocardial infarction-associated transcript (MIAT) by I₂ were directly associated with decreased MYCN and TrkB expression. In contrast, PPARγ activation was accompanied by overexpression of FasN and TrkA and a significant decrease in Aurka, a MYCN-stabilizing protein. In zebrafish, I₂-pretreated SK-N-BE(2) xenografts exhibited a clear reduction in angiogenesis (vascular density) and a decrease in invasive capacity. In conclusion, I₂ supplementation decreases cell viability and attenuates invasion and angiogenesis in NB cells, highlighting its potential as an adjuvant to conventional therapy for high-risk NB. Full article

This study aimed to investigate the molecular mechanisms underlying dopaminergic injury induced by gestational and lactational atrazine (ATR) exposure in rat offspring, with a particular focus on non-coding RNA-mediated regulation. Pregnant rats were exposed to ATR during gestation and lactation. Offspring underwent behavioral testing at postnatal day 21 (PND21) and were sacrificed for midbrain tissue collection at PND28. Behavioral alterations, histopathological changes in the substantia nigra, and dopaminergic marker expression were assessed to evaluate ATR-induced neurotoxicity. Whole-transcriptome sequencing was then performed to identify differentially expressed mRNAs, miRNAs, and lncRNAs, followed by co-expression, protein–protein interaction, and competing endogenous RNA (ceRNA) network analyses. Key targets were validated by qRT-PCR. Candidate molecules identified from transcriptomic and ceRNA analyses were further examined in an ATR-induced neurotoxicity model established in RA-differentiated SK-N-SH cells. Dual-luciferase reporter, Ago2-RNA immunoprecipitation, and biotin-labeled RNA pull-down assays were used to examine putative binding relationships and molecular interactions. In addition, lentivirus-mediated Elavl4 overexpression was performed to further evaluate the role of this candidate regulator in ATR-induced Nurr1 downregulation. Gestational and lactational ATR exposure induced significant behavioral abnormalities in rat offspring. These changes were accompanied by histopathological alterations in the substantia nigra, including reduced TH immunoreactivity, as well as abnormal expression of dopaminergic markers, characterized by decreased TH and Nurr1 levels and increased α-syn expression. Together, these findings indicate the presence of dopaminergic injury. Whole-transcriptome analysis further revealed widespread dysregulation of mRNAs, miRNAs, and lncRNAs in ATR-exposed offspring. Subsequent integrative analysis suggested a potential ceRNA regulatory relationship among Elavl4, miR-301a-5p, and Nurr1, which was further supported by qRT-PCR. Dual-luciferase reporter, RIP, and RNA pull-down assays supported direct interactions between miR-301a-5p and both Elavl4 and Nurr1, as well as their association with the Ago2-containing silencing complex. Moreover, Elavl4 overexpression partially reversed ATR-induced Nurr1 downregulation in vitro. Gestational and lactational ATR exposure induced behavioral abnormalities and dopaminergic injury in rat offspring. Whole-transcriptome analysis combined with experimental validation suggests a potential association between the Elavl4/miR-301a-5p/Nurr1 ceRNA axis and ATR-induced dopaminergic injury, providing insight into the post-transcriptional mechanisms underlying developmental neurotoxicity. Full article

The Skewed5 Similar (SKS) genes play a crucial role in plant growth and development, as well as in responding to abiotic stress, by regulating cell wall remodeling and maintaining reactive oxygen species (ROS) homeostasis. The SKS gene family has not yet been thoroughly studied in soybean. We conducted a comprehensive genome-wide analysis of 88 GmSKS genes, systematically elucidating their gene structures, conserved protein domains, collinearity relationships, and phylogenetic relationships, to identify potential candidate genes associated with soybean tolerance to salt stress. The GmSKS genes are distributed across 18 chromosomes, and the expansion of this gene family is primarily attributed to the combined effects of tandem duplications and segmental duplications. Different tissue-specific expression patterns among GmSKS members were identified using expression profiling. Analysis of cis-regulatory elements further revealed that the promoter region may be involved in plant hormone signaling pathways and responses to abiotic stress. Furthermore, quantitative reverse transcription PCR (qRT-PCR) analysis showed that 21 of the 22 examined GmSKS genes were significantly upregulated under salt stress, while one was significantly downregulated. This expression pattern may be linked to salt tolerance mechanisms in soybean under stress conditions. Haplotype and selection trend analyses of GmSKSs revealed that varieties carrying GmSKS5^1123G, GmSKS22^1727G, GmSKS43^50T and GmSKS71^1213T are highly enriched in cultivated soybeans and have undergone artificial selection. This study provides basic information for the identification of salt stress-responsive gene resources of GmSKS family genes, and provides novel theoretical insights for the functional identification and cloning of soybean salt tolerance-related genes. Full article

Neonicotinoid insecticides and fluoroquinolone antibiotics frequently co-occur in aquatic and terrestrial environments, posing a threat to human health, yet their combined neurotoxic potential remains poorly characterized. This study aimed to assess the cytotoxicity of typical neonicotinoids and fluoroquinolones as well as their mixtures in human neuroblastoma SK-N-SH cells and identify affected pathways. SK-N-SH cells were exposed to clothianidin (CLO), imidacloprid (IMI), enrofloxacin (ENR), and ofloxacin (OFX) individually and in fixed-ratio mixtures (50% of each compound’s IC₅₀) for 24 h and 48 h, and cell viability was quantified using the alamarBlue^® method. Single-compound dose–response testing showed time-dependent cytotoxicity, with higher potency for fluoroquinolones (24 h IC₅₀: ENR 1.446 mM, OFX 2.742 mM; 48 h IC₅₀: ENR 0.826 mM, OFX 2.005 mM) than neonicotinoids (24 h IC₅₀: IMI 4.754 mM, CLO 5.356 mM; 48 h IC₅₀: IMI 3.631 mM, CLO 4.029 mM). Concentration-addition analysis indicated that most mixtures produced synergistic interaction in reduction in cell viability, with ENR+OFX showing the strongest effect at 48 h (Observed viability 7.138% vs. Predicated viability 82.368%). RNA-seq (24 h) revealed that binary mixtures generally induced more differentially expressed genes than single exposures, and ENR-containing mixtures showed the largest transcriptomic shifts, enriching pathways related to cellular stress and injury as well as neuronal signaling and connectivity. RT-qPCR validated the changes in expressions of five key neurobiology-relevant genes (LMO3, NOS1, ADCY8, FGF7 and TNFRSF12A). These findings highlight the importance of assessing insecticide–antibiotic mixtures when evaluating their hazards in environment. Full article

Airborne particulate matter with a diameter of <10 μm (PM₁₀) can damage the corneal epithelium by inducing oxidative stress, disrupting the NRF2 antioxidant pathway, and triggering epithelial barrier dysfunction and inflammation. However, the role of mitochondria in mediating PM₁₀-induced damage remains unexplored. This study investigated the impact of PM₁₀ on mitochondrial homeostasis in both immortalized human corneal epithelial cells (HCE-2) and the mouse corneal epithelium, as well as the protective effects of SKQ1. For in vivo assessment, female C57BL/6 mice were exposed to either control air or PM₁₀ (±SKQ1) in a whole-body exposure chamber for 2 weeks (3 h/day, 5 days/week, with weekends off). In vitro, HCE-2 cells were exposed to 100 μg/mL PM₁₀ (±SKQ1) for 24 h, and mitochondrial function and morphology were evaluated. In vitro, PM₁₀ significantly impaired mitochondrial function by reducing basal, maximal, and ATP-linked respiration; reserve capacity; and coupling efficiency compared to the control and SKQ1 groups. PM₁₀ also downregulated mitofusin1 (MFN1) and optic atrophy1 (OPA1) and upregulated dynamin-related protein1 (DRP1) and mitochondrial fission protein1 (FIS1) in HCE-2 cells. In addition, PM₁₀ exposure significantly decreased the mitochondrial membrane potential; mitochondrial DNA copy number; and cytochrome c oxidase subunit 4 isoform 1 (COX4i1), mitochondrial transcription factor A (TFAM), and peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) levels. SKQ1 pre-treatment significantly attenuated these effects. In vivo, PM₁₀ exposure significantly decreased the levels of MFN1, TFAM, COX4i1, and superoxide dismutase (SOD2), whereas SKQ1 treatment significantly reversed these effects. Overall, these findings demonstrate that PM₁₀ exposure induces mitochondrial fragmentation, disrupts mitochondrial biogenesis and quality control, and reduces mitochondrial respiration, resulting in mitochondrial dysfunction. SKQ1 effectively reversed these changes, suggesting its potential as a therapeutic strategy to protect corneal epithelial cells from PM₁₀-induced mitochondrial damage. Full article

Glycogen synthase kinase 3 (GSK3/Shaggy-like) is a highly conserved serine/threonine kinase that orchestrates growth, hormone signaling, and abiotic stress responses in both animals and plants, yet its role in watermelon remains unexplored. In this study, we conducted a whole-genome identification, identifying a total of eight members of the GSK3 gene family (ClGSK3) distributed across seven chromosomes. Phylogenetic and synteny analyses resolved the eight ClGSK3s into four subfamilies that display one-to-one or one-to-many orthology with Arabidopsis and rice GSK3 genes, indicating conserved genomic micro-collinearity across dicots and monocots. Predictions of cis-acting elements and transcriptome data analysis indicate that ClGSK3s may be involved in hormone- and stress-responsive conditions. Protein–protein interaction networks predicted 53 candidate partners for five ClGSK3 proteins; yeast two-hybrid assays subsequently confirmed that ClSK21 associates with three of them—orthologs of the core brassinosteroid (BR)-signaling components BKI1 and BZR1. qRT-PCR revealed that ClSK21, ClSK31, and ClSK41 are rapidly and significantly reprogrammed by BR treatment. Collectively, our data suggest that ClGSK3s modulate fruit development and stress tolerance by integrating hormone-related pathways, especially BR signaling. Future studies are encouraged to integrate genetics and multi-omics approaches to systematically validate the roles of ClGSK3s in hormone signaling and abiotic stress responses. Full article

Background/Objectives: Neuroblastoma cells are phenotypically plastic, transitioning between mesenchymal and adrenergic states. Core functional genes (e.g., YAP1) mark the mesenchymal state, which is linked to unfavorable prognosis. We and others previously demonstrated relapse-specific Hippo-YAP pathway activation in matched primary/relapsed neuroblastomas. Here we explored the role of YAP1 in neuroblastoma aggressiveness and response to therapy. Methods: RT-qPCR and immunoblotting assessed YAP1 expression in neuroblastoma cell lines. RNA-sequencing detected YAP1-dependent gene signatures in Tet-ON SK-N-AS and SH-EP neuroblastoma cell models expressing wildtype YAP1 or constitutively activated YAP1^S127A. Data from cell models were compared with our published YAP1 expression data from neuroblastomas. Efficacy of commonly used chemotherapeutics was comparatively analyzed in the cell models. Results: YAP1 expression showed marked variability across a panel of neuroblastoma cell lines, assessed by mRNA analysis in 10 cell lines and protein analysis in a subset of 9 cell lines. RNA sequencing in constitutively activated YAP1^S127A mutant and wildtype YAP1 models detected 2162 and 1837 significantly differentially expressed genes in the SK-N-AS and SH-EP backgrounds, respectively. Continuously activating YAP1 in SK-N-AS cells upregulated mesenchymal signature genes and mesenchymal-associated transcription factors. Gene expression influenced by YAP1 activity in the cell models significantly overlapped with YAP1-associated genes (e.g., CYR61 and SPRY4) in published tumor data. Functionally, YAP1^S127A expression rendered neuroblastoma cells resistant to chemotherapy. Conclusions: Findings corroborate the idea of a mechanistic role for YAP1 in neuroblastoma adrenergic to mesenchymal reprogramming and therapy resistance. The YAP1-mediated plastic switch towards a mesenchymal expression state in neuroblastoma cells may provide the molecular basis for novel therapeutic avenues. Full article

Aging and neurodegenerative diseases are characterized by common features involving bioenergetics deficiencies, oxidative stress and alterations of calcium buffering. Mechanisms of mitochondrial-targeted drugs include the modulation of electron transport chain and oxidative phosphorylation, the binding to mitochondrial lipids, free-radical scavenging, calcium signaling, and possible effects on mitochondrial biogenesis and dynamics and on the regulation of mitophagic pathways. One of the main sites of action of mitochondria-targeted drugs is the interaction with respiratory chain components. Mitochondrial-targeted compounds such as Mito-Q, and Mito-apocynin have been developed by conjugating triphenylphosphonium (TPP⁺) lipophilic cation group with natural molecules, therefore obtaining promising drugs for reestablishing the correct functioning of the mitochondrial respiratory chain. Stabilization of cardiolipin at the inner mitochondrial membrane by elamipretide or SkQ1 and mitochondria-targeted ROS scavengers can also offer a therapeutic approach to prevent bioenergetic impairment associated with several diseases. In addition, the modulation of calcium signaling can be achieved using both MCU agonists and antagonists representing another mitochondrial target for drug therapies development. Finally, potential strategies for treating neurodegenerative diseases based on the modulation of mitochondrial biogenesis, dynamics and/or mitophagic pathways are discussed. Full article

Vietnam faces a hyperendemic burden of hepatitis B virus (HBV) infection, but the prevalence of occult HBV infection (OBI) and its underlying molecular mechanisms in healthy populations remain poorly understood. This study aimed to characterize the serological and molecular HBV profile of a healthy Vietnamese adult cohort in Southern Vietnam. We assessed the prevalence of occult HBV infection (OBI) and HBsAg-positivity (serving as a proxy for probable chronic infection). In this cross-sectional study, 397 healthy adults from Southern Vietnam underwent serological screening for HBsAg, anti-HBs, and anti-HBc. All participants were screened for HBV DNA using a high-sensitivity PCR assay (LOD ≥ 5 IU/mL). For all viremic cases, the full Pre-S/S region was sequenced to determine genotype and characterize escape mutations. We uncovered a high prevalence of both HBsAg-positivity (17.6%) and OBI (9.3% HBsAg-negative, HBV DNA-positive). Serological analysis revealed a massive, age-dependent reservoir of past exposure (63.7% anti-HBc) characterized by a high and increasing prevalence of the anti-HBc only profile (31.5%), a key serological marker for OBI. This trend contrasted sharply with a steep age-related decline in protective anti-HBs. The viral landscape was dominated by genotypes B (73.8%) and C (26.2%), with sub-genotypes B4 and C1 being the most prevalent. Critically, individuals with OBI carried a significantly higher burden of S gene escape mutations compared to those with HBsAg-positivity (p < 0.001). Canonical escape variants, including sG145R (21.6%), sK141R/T/E/Q (24.3%), and sT116N/A/I/S (18.9%), were exclusively or highly enriched in the OBI group. A LASSO-logistic model based on this mutational profile successfully predicted occult infection with high accuracy (AUC = 0.83). A substantial hidden reservoir of occult HBV infection exists within the healthy adult population of Vietnam, driven by a high burden of S gene escape mutations. These findings highlight the significant limitations of conventional HBsAg-only screening. They also underscore the need for comprehensive molecular surveillance to address the true scope of HBV viremia, hopefully enabling a reduction in hidden transmission of clinically significant viral variants. Full article

Mitochondrial dysfunction and oxidative stress are crucial contributors to the pathogenesis of Alzheimer’s disease (AD) and dementia exhibiting cognitive decline at the early stage of neurodegeneration. Natural vitamin antioxidants (NVAs) and novel mitochondria-targeted antioxidants (MTAs) are proposed as potential therapeutics though conclusive evidence is lacking. Objectives were to examine in vivo evidence on NVAs and MTAs for preventing and/or treating cognitive decline leading to dementia, to identify the most promising antioxidants, and highlight translational gaps. Methods followed PRISMA-ScR guidelines. MEDLINE, EMBASE and Scopus were searched for English language in vivo experiments assessing NVAs or MTAs in AD and dementia. A total of 25 studies (13 NVAs; 12 MTAs) met inclusion criteria. NVAs (Vitamin A, B, C, E) demonstrated mixed efficacy in reducing oxidative stress and improving cognitive outcomes, with Vitamin E showing the most consistent neuroprotective effects. MTAs (MitoQ, MitoTEMPO, SS31, SkQ1) improved mitochondrial dynamics and cognitive performance and reduced dementia-related pathology. Both NVAs and MTAs improved biomarker profiles and cognitive outcomes in vivo animal models of AD and dementia, but MTAs showed more robust and consistent efficacy by directly targeting mitochondrial pathways. Given the favourable safety profiles of MTAs in other clinical conditions, early-phase human trials in dementia and AD are warranted to evaluate their long-term cognitive benefits. Full article

During the storage of boar sperm at 17 °C, reactive oxygen species (ROS) are continuously generated. Excessive ROS can disrupt the mitochondrial redox balance and cause sperm damage. In this study, boar semen was diluted with solutions containing different concentrations of the natural antioxidant lycopene (1, 5, 10, 20, 40, 80 μM) or the mitochondria-targeted antioxidant SKQ1 (1, 5, 10, 25, 50, 70 nM), and sperm vitality was assessed throughout storage at 17 °C. Based on the screening results, the optimal concentrations were selected for combined application to investigate their effects on sperm quality and potential synergistic interactions. The results demonstrated that sperm motility was significantly higher in the 20 μM lycopene and 50 nM SKQ1 treatment groups compared to the control (p < 0.05). The combined treatment of 20 μM lycopene and 25 nM SKQ1 exhibited a synergistic effect, significantly improving sperm vitality, acrosome and membrane integrity, superoxide dismutase (SOD), glutathione peroxidase (GSP), adenosine triphosphate (ATP) levels (p < 0.05). Meanwhile, ROS and malondialdehyde (MDA) levels were significantly reduced (p < 0.05). Metabolomics analysis identified 52 differential metabolites (p < 0.05), including ABC transporters, corticosterone, and palmitic acid. KEGG pathway enrichment analysis revealed that these metabolites were mainly associated with steroid hormone biosynthesis, ABC transporters, and AMPK signaling pathways (p < 0.05), most of which were related to sperm cell energy metabolism and signal transduction. Furthermore, treatment with antioxidants significantly increased p-AMPK and Nrf2 expression in sperm cells (p < 0.05). These findings suggest that the combination of lycopene and SKQ1 improves boar sperm quality during 17 °C storage by enhancing energy metabolism and mitigating oxidative stress, potentially through the activation of the AMPK/Nrf2 pathway. Full article