Search Results (1,225)

Lipopolysaccharide (LPS) impairs the function of ovine follicular granulosa cells (GCs), representing a primary cause of follicular atresia. Selenium (Se), an essential trace element, possesses anti-inflammatory and cytoprotective properties; however, its effects on GC ultrastructure remain largely unknown. In this study, primary ovine GCs were exposed to LPS (10 µg/mL) and treated with sodium selenite (25 nM). Transmission electron microscopy (TEM), JC-1 staining, enzyme-linked immunosorbent assay (ELISA), reactive oxygen species (ROS) detection, flow cytometry, and quantitative real-time PCR (qRT-PCR) were employed to evaluate cellular ultrastructure, mitochondrial membrane potential (ΔΨm), and downstream physiological processes. LPS induced severe mitochondrial pyknosis, cristae loss, and reduced ΔΨm, accompanied by inflammation, oxidative stress, apoptosis, and impaired steroidogenesis. Se intervention markedly ameliorated these ultrastructural injuries, preserving mitochondrial morphology and ΔΨm. Functionally, Se suppressed the release of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β); enhanced the activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) while attenuating ROS accumulation; inhibited apoptosis by upregulating BCL-2 and downregulating BAX and CASPASE-3; and restored E₂ and P₄ secretion via upregulation of STAR and NR5A1. This study provides direct morphological evidence that Se protects ovine GCs from LPS-induced damage by repairing mitochondrial ultrastructure. This structural restoration is central to its integrated anti-inflammatory, antioxidant, anti-apoptotic, and steroidogenic effects. These in vitro findings suggest that Se may serve as a promising nutritional strategy for mitigating inflammation-driven follicular atresia, pending further in vivo validation. Full article

Background/Objectives: Drug repurposing has emerged as a promising strategy for identifying novel anticancer agents among clinically established drugs. Fexofenadine, a second-generation H1 antihistamine, has been proposed as a candidate for repurposing in oncology; however, the molecular mechanisms underlying its biological activity remain insufficiently characterized. This study investigated the effects of fexofenadine on oxidative stress, mitochondrial function, apoptosis, and pro-survival signaling pathways in cervical and lung cancer cells. Methods: HeLa and A549 cancer cells, as well as non-tumorigenic Beas-2B epithelial cells, were exposed to fexofenadine under in vitro conditions. Cell viability, apoptosis, reactive oxygen species generation, mitochondrial membrane potential, DNA damage, autophagy-associated responses, and PI3K/AKT and MAPK/ERK pathway activation were assessed using flow cytometry, fluorescence microscopy, electron microscopy, and biochemical assays. Three-dimensional spheroid cultures and N-acetyl-L-cysteine rescue experiments were additionally employed to evaluate biological relevance and the contribution of oxidative stress. Results: Fexofenadine induced concentration-dependent accumulation of reactive oxygen species, mitochondrial membrane depolarization, Bcl-2 inactivation, caspase-3/7 activation, DNA damage, and apoptotic cell death in HeLa and A549 cells. Antioxidant pretreatment with N-acetyl-L-cysteine significantly reduced oxidative stress, attenuated mitochondrial dysfunction, and partially suppressed apoptosis. Fexofenadine was associated with reduced PI3K/AKT and MAPK/ERK pathway activation and promoted autophagy-associated responses. In three-dimensional spheroid cultures, treatment disrupted spheroid integrity and increased apoptotic cell death. Non-tumorigenic Beas-2B cells exhibited lower sensitivity to treatment than malignant cells. Conclusions: Fexofenadine disrupts redox homeostasis and is associated with reduced activation of pro-survival signaling pathways, resulting in oxidative stress-associated mitochondrial dysfunction and apoptosis in cancer cells. These findings provide mechanistic support for further evaluation of fexofenadine as a candidate for anticancer drug repurposing, while additional pharmacokinetic and in vivo studies are required to determine its translational relevance. Full article

Epilepsy is a common chronic neurological disorder characterized by recurrent seizures. Gastrodia elata, the dried tuber of G. elata Bl. (Orchidaceae), is a valuable medicinal and edible botanical resource. This study optimized the preparation of Yellow Rice Wine-Processed G. elata (YPGE) and investigated its antiepileptic effects and underlying mechanisms in a pentylenetetrazol (PTZ)-kindled rat model. Processing parameters were optimized using single-factor experiments combined with an analytic hierarchy process (AHP)-entropy weight method (EWM) weighting strategy and Box–Behnken design–response surface methodology. The optimal parameters were determined as 18% alcohol by volume, 72 °C drying temperature, and 32 h drying time. Compared with unprocessed G. elata (GE), YPGE exhibited 0.54-, 0.13-, 1.87-, and 3.58-fold increases in the contents of gastrodin (GAS), G. elata polysaccharides (GEPs), p-hydroxybenzyl alcohol (p-HBA), and total parishins (TP), respectively, and demonstrated significantly enhanced in vitro antioxidant activity (IC50 values of 2.604, 2.719, and 4.046 mg/mL for DPPH, ABTS, and hydroxyl radicals). In vivo, both GE and YPGE significantly reduced seizure severity, decreased inflammatory cytokines (TNF-α, IL-1β), alleviated oxidative stress (increased SOD and GSH-Px, decreased MDA), and modulated neurotransmitter balance (reduced Glu, increased GABA) in brain tissues. YPGE also upregulated P-glycoprotein expression and reduced neuronal apoptosis in the hippocampal CA1 region by upregulating Bcl-2 and downregulating Bax. These findings suggest that YPGE exerts multi-target antiepileptic effects through synergistic anti-inflammatory, antioxidant, and anti-apoptotic actions, providing experimental evidence for the development of novel antiepileptic therapies based on processed G. elata. Full article

We aimed to investigate the therapeutic effects of alpha-pinene (α-pinene), a major component of phytoncide, in in vitro and in vivo models of mild traumatic brain injury (mTBI), raising the possibility of treating mTBI, a condition currently lacking adequate therapies. An in vitro model was established using SH-SY5Y cells and a cell injury controller, and was treated with α-pinene (0.5 g/mL). An in vivo model was induced by a stereotaxic impactor in male C57BL/6J mice and treated with α-pinene intravenously (50 mg/kg and 100 mg/kg) for 3 days post-injury. Histopathological and immunohistochemical comparisons were conducted alongside cognitive function tests to evaluate -pinene treatment. In vitro analysis showed that alpha-pinene treatment significantly increased TUNEL-positive cells. Elevated NOX4 and p22phox mRNA expressions and a high Bax/Bcl-2 protein expression ratio were noted following alpha-pinene treatment. mTBI mice treated with alpha-pinene exhibited a notable decrease in brain water content with fewer FJB-positive neurons and lower protein expression of Bax and Bcl-2 compared to untreated mTBI mice. Immunofluorescence staining for NOX4 and GFAP-positive or Iba-1-positive cells demonstrated that the increased oxidative stress and astrogliosis or activated microglia triggered by mTBI were alleviated after alpha-pinene treatment. Cognitive function testing revealed a general improvement in mTBI mice treated with alpha-pinene, with statistical significance observed in the NOR test. Alpha-pinene appears to be beneficial for neuroprotection and enhancing cognitive function in the early phases of mTBI. Full article

Marine medicinal algae represent a valuable reservoir of bioactive metabolites for anticancer drug discovery, yet the efficient identification of target-relevant compounds from chemically complex marine matrices remains challenging. In this study, an integrated cathepsin B-oriented strategy was developed to discover, prioritize, isolate, and validate antitumor metabolites from the brown alga Sargassum polycystum. Affinity ultrafiltration LC-MS was first applied to screen CTSB-binding constituents from the crude extract, followed by molecular docking, molecular dynamics simulation, and gray relational analysis for multidimensional candidate prioritization. Seven CTSB-binding metabolites were characterized, including chlorogenic acid, caffeic acid, cynarin, loliolide, taxifolin, senkyunolide H, and dihydroactinidiolide, with binding degrees of 73.99–85.61% at 2.5 U/mL CTSB. Molecular docking showed predicted binding affinities ranging from −6.3 to −9.4 kcal/mol, compared with −10.2 kcal/mol for the positive control CA-074Me. Integrated computational and biological evaluation identified caffeic acid, cynarin, and taxifolin as the top-ranked candidates. Preparative recovery was then achieved using counter-current chromatography combined with semi-preparative HPLC, and the isolated compounds were structurally identified by LC-MS/MS and NMR. Cellular assays in NCI-H1975 cells suggested that these metabolites reduced CTSB-associated enzymatic activity and intracellular CTSB-related fluorescence signals to different extents, with phenolic acid-type compounds exhibiting comparatively stronger effects. At the extract level, S. polycystum dose-dependently suppressed NCI-H1975 xenograft tumor growth, with inhibition rates of 48.78%, 36.58%, and 22.86% in the high-, middle-, and low-dose groups, respectively, without evident hepatorenal histopathological toxicity. This effect was associated with reduced CTSB, Ki-67, and Bcl-2 staining, increased Bax staining, enhanced apoptosis, and ultrastructural alterations in tumor tissues. Overall, this study provides a practical CTSB-oriented workflow for discovering antitumor metabolites from marine medicinal algae and supports further investigation of S. polycystum as a potential source of anti-NSCLC candidates. Full article

Carnosine is a naturally occurring endogenous dipeptide with great potential to improve reproductive function and fertility. In this study, supplementation of 1 μg/mL carnosine during in vitro maturation (IVM) significantly enhanced the developmental competence and quality of the resulting bovine embryos. Carnosine treatment effectively elevated mitochondrial membrane potential, mitochondrial activity, and ATP content in oocytes. Moreover, it strengthened the antioxidant and anti-apoptotic capacities of oocytes, as evidenced by reduced intracellular reactive oxygen species (ROS) levels, lowered DNA damage and an early apoptosis rate, alongside increased glutathione (GSH) content, an elevated BCL2/BAX mRNA ratio, and upregulation of antioxidant genes SOD1, CAT, GPx1, and GPx4. Notably, combined application of 1 μg/mL carnosine during IVM and 10⁻⁷ M melatonin during in vitro culture (IVC) synergistically improved both blastocyst development and quality. Collectively, these findings provide novel evidence supporting the therapeutic potential of carnosine in optimizing in vitro embryo production in bovine, and highlight the value of stage-specific supplementation strategies to further improve embryonic development efficiency. Full article

Background: Rosmarinus officinalis L. is a medicinally important aromatic plant rich in bioactive secondary metabolites with diverse therapeutic properties. This study aimed to characterize the phytochemical profile of R. officinalis leaf extracts, isolate carnosic acid as a major bioactive diterpene, and evaluate its biological activities. Methods: Leaf extracts were prepared using solvents of increasing polarity and analyzed by phytochemical screening and UHPLC/QTOF-MS. Carnosic acid was isolated by thin-layer chromatography and assessed for antibacterial, antibiofilm, antioxidant, anti-inflammatory, antidiabetic, and antiproliferative activities using in vitro assays. Expression of apoptosis-related genes was also investigated. Results: Methanolic and ethanolic extracts exhibited the highest abundance of phenolic compounds and secondary metabolites, whereas the hexane extract showed lower phytochemical content. UHPLC/QTOF-MS identified seven major metabolites, including phenolic acids, flavonoids, and abietane-type diterpenes. Purified carnosic acid demonstrated potent antibacterial activity (MIC: 10–23 μg/mL) and inhibited biofilm formation by up to 90%. Strong antioxidant activity was observed, with DPPH and ABTS radical-scavenging IC₅₀ values of 125 and 130 μg/mL, respectively. The compound also exhibited notable anti-inflammatory activity and markedly inhibited α-amylase and α-glucosidase activities. Furthermore, carnosic acid exhibited dose-dependent antiproliferative activity against MCF-7, HepG2, and MCF-10A cells, reducing cell viability to 10.8%, 16.9%, and 70.4 ± 1.8%, respectively, at 250 μg/mL, with corresponding IC₅₀ values of 28.3, 37.8, and >250 μg/mL, respectively. Gene expression analysis revealed upregulation of BAX and downregulation of BCL2, indicating activation of mitochondrial-mediated apoptosis. Conclusions:R. officinalis leaves represent a valuable source of multifunctional phytochemicals, particularly carnosic acid. Its broad-spectrum biological activities and apoptosis-inducing potential support its promising application in pharmaceutical, nutraceutical, and biomedical fields. Full article

The Houpoea officinalis flower (HOF) represents an underutilized sustainable bio-resource. This study systematically evaluated its potential using an ethanol-based green extraction process optimized by Response Surface Methodology, with the optimal conditions consisting of approximately 50% ethanol, a solvent-to-solid ratio of 54 mL/g, and an extraction time of 31 min. Chemical profiling across four developmental stages—S1 (Bud), S2 (Bud swelling), S3 (Initial flowering), and S4 (Full bloom)—suggested magnolol and honokiol as the major phenolic compounds, showing a trend of decline during early development followed by an increase at the S4 stage. A significant positive correlation was observed between total phenolic content and antioxidant activity, and the S1 stage extract displayed the strongest antioxidant capacity in multiple in vitro assays. Network pharmacology analysis predicted oxidative stress-related targets and pathways, with TP53, AKT1, IL6, BCL2, and CASP3 recognized as key hub genes. Molecular docking further predicted favorable binding interactions between major HOF phenolics and these target proteins. Collectively, these findings reveal the multi-target antioxidant potential of HOF and provide evidence supporting its potential role in antioxidant-related traditional applications based on predicted mechanisms. Moreover, HOF, particularly at the S1 developmental stage, shows promise as a sustainable source of natural antioxidants and functional ingredients, promoting the high-value utilization of agricultural by-products. Full article

Cancer remains the foremost cause of death globally and presents a major obstacle to increasing life expectancy. The identification of natural anticancer agents is therefore a key research priority. Essential oils (EOs), widely used in traditional medicine, possess diverse biological activities that warrant systematic investigation. Anisosciadium orientale is traditionally regarded as a safe, edible herb; however, its therapeutic potential has not been extensively explored. In this study, the chemical profile of the EO obtained from the aerial parts of A. orientale was characterized using GC–MS analysis. Antioxidant activity was evaluated through hydrogen peroxide and ABTS radical-scavenging assays, whereas anticancer effects and underlying mechanisms were evaluated in multiple cancer cell lines using MTT cytotoxicity assays, flow cytometry, and molecular docking studies. Sixty constituents were identified in the EO, with myristicin and its isomer among the major components. The EO exhibited notable antioxidant and anticancer activities, demonstrating cytotoxicity against lung carcinoma A549 cells with an IC₅₀ of 84.8 µg/mL and inducing significant apoptosis accompanied by G₂/M cell-cycle arrest. Treatment with the EO markedly boosted levels of caspase-3, p53, Bax, and the Bax/Bcl-2 ratio, while downregulating Bcl-2 expression. Molecular docking revealed strong binding affinities of major constituents—particularly myristicin and its isomer—toward the EGFR kinase active site, suggesting a high degree of complementarity with the EGFR kinase domain. Collectively, this study represents the first comprehensive study integrating chemical profiling, in vitro cytotoxicity, mechanistic assays, and molecular docking for A. orientale. These findings position A. orientale EO as a promising scaffold for the development of natural anticancer interventions, providing a foundation for future preclinical exploration. Full article

Background: Apoptosis is a fundamental process for maintaining tissue homeostasis, and its dysregulation is closely linked to the development of numerous diseases, including colorectal cancer. In recent years, dietary polyphenols have gained interest due to their antioxidant, pro-apoptotic, and chemopreventive properties. Artichoke (Cynara scolymus L.) by-products are rich source of hydroxycinnamic acids and flavonoids, making them promising source of bioactive compounds. Methods: In this study we evaluated the cytotoxic and pro-apoptotic activity of four aqueous extracts obtained from artichoke bract by-products, including one commercial hybrid (CAPB) and three local Apulian varieties (BriB, VaMB, LMTB), in human colorectal adenocarcinoma cell lines (Caco-2 and HT29). The extracts were characterized according to their total polyphenol content and phenolic profile. Results: The selected artichoke by-product extracts exhibited significant cytotoxic effects both in a concentration- and time-dependent manner, with concentrations ≥ 2 mg/mL significantly reducing cell viability and nearly abolishing it at 4 mg/mL after 48 h. Moreover, treatment with the extracts modulated the expression of apoptosis-related proteins, characterized by an increase in pro-apoptotic markers (Bax, caspase-9, caspase-3) and a decrease in the anti-apoptotic protein Bcl-2, suggesting activation of the mitochondrial apoptotic pathway. In particular, the BriB extract was able to induce an apoptosis rate higher than 80% in Caco-2 cells and achieved comparable rates in HT29 cells at concentrations of 2–3 mg/mL. Conclusions: Overall, these findings demonstrate that artichoke by-product extracts exert significant pro-apoptotic effects in colorectal cancer cells and highlight their potential as sustainable sources of bioactive compounds for nutraceutical or adjuvant anticancer applications. Full article

New copper(II) (C1) and palladium(II) (C2) complexes with S,O-tetradentate ligand (L) derived from thiosalicylic and thiopropionic acids were synthesized. In cell-based assays, (C1) exhibited the most pronounced activity within the tested compound series and was therefore advanced for mechanistic evaluation in 4T1 triple-negative breast cancer cells. (C1) significantly reduced 4T1 cell viability by inducing early and late apoptosis, accompanied by mitochondrial membrane depolarization and enhanced cytochrome C release. Consistently, (C1) increased the Bax/Bcl-2 ratio, promoting a pro-apoptotic shift. In parallel, (C1) triggered autophagy, as evidenced by decreased p62 and LC3B levels, induced G0/G1 cell-cycle arrest, and suppressed proliferative signaling by downregulating Ki67, cyclin D, and phosphorylated AKT. The DNA-binding studies showed moderate to strong affinity, favoring minor groove binding, with higher affinity for (C1) than for (C2). Tryptophan fluorescence quenching indicated a strong interaction with BSA via a predominantly static mechanism, more pronounced for (C1). Molecular docking at the DNA and BSA binding sites corroborated experimental findings and suggested favorable interactions between the complexes and apoptosis-related proteins (CASP3, BAX, and BCL2). The integrated experimental and computational data identify (C1) as a biologically active compound with multimodal biological effects in vitro, supporting further structural optimization and mechanistic investigation. Full article

Global freshwater salinization endangers aquatic species, yet its impacts on crustaceans remain poorly understood. This study investigated the hepatopancreatic response of Eriocheir sinensis to carbonate alkalinity stress (0, 4.375, 8.75, 17.5, and 35 mmol/L) over 24, 48, and 96 h, integrating histology, ultrastructure, gene expression (RT-qPCR), and non-specific immune enzyme assays. Histopathological and ultrastructural analyses revealed concentration- and time-dependent damage, including vacuolization, hepatic tubule disintegration, nuclear condensation, mitochondrial reduction, and loss of cellular integrity. Molecular analysis demonstrated upregulation of genes associated with the TLR2-MyD88-NF-κB pathway and inflammatory genes (LITAF, IL-16), alongside increased HSP70 expression, confirming severe inflammation and cellular stress. Furthermore, apoptosis was induced via upregulated Bax and Caspase-3, downregulated Bcl-2, and DNA fragmentation. Non-specific immune responses in the hepatopancreas exhibited dynamic changes: acid phosphatase (ACP) was initially activated at low alkalinity but inhibited at high concentrations, while alkaline phosphatase (AKP) activity increased at 96 h. Notably, the hepatopancreas proved more sensitive to this stress than the hemolymph. Collectively, carbonate alkalinity causes multidimensional hepatopancreatic injury in E. sinensis through structural disruption, inflammation mediated by TLR2-MyD88-NF-κB signaling pathway-related genes, apoptosis induction, and immune enzyme dysregulation, posing a significant threat to crab health in salinized waters. Full article

Melanoma exhibits pronounced metabolic plasticity and mitochondrial dependency, contributing to therapeutic resistance and tumor progression. Targeting mitochondrial function therefore represents a promising anticancer strategy. 2-Aminoethyl dihydrogen phosphate (2-AEH₂P), a bioactive phosphomonoester, has demonstrated antiproliferative potential, while metformin, a clinically established antidiabetic drug, acts as a mitochondrial complex I inhibitor and metabolic modulator. This study investigated the cytotoxic and mechanistic effects of 2-AEH₂P and metformin hydrochloride, individually and in combination, in human (SK-MEL-28) and murine (B16-F10) melanoma models, using non-tumorigenic fibroblasts (FN1 and L929) as controls. Cell viability, proliferation dynamics, cell-cycle distribution, mitochondrial membrane potential (ΔΨm), and apoptosis-associated markers were evaluated by flow cytometry. 2-AEH₂P reduced melanoma cell viability and proliferation while inducing G2/M accumulation, DNA fragmentation, mitochondrial depolarization, increased cytochrome c release, caspase-3 and caspase-8 activation, upregulation of p53 and Bad, and downregulation of Bcl-2. Metformin alone exerted moderate cytotoxic and pro-apoptotic effects. Notably, combined treatment markedly potentiated mitochondrial depolarization and intrinsic apoptotic signaling in melanoma cells, significantly lowering IC₅₀ values and enhancing caspase activation and cytochrome c release. Bliss independence analysis demonstrated synergistic interaction in SK-MEL-28 and B16-F10 cells. Although interaction scores indicated synergy in one fibroblast model, absolute cytotoxicity remained lower than in melanoma cells. These findings demonstrate that metabolic co-targeting with metformin enhances mitochondrial dysfunction-associated apoptotic signaling in melanoma cells, supporting a drug repositioning strategy aimed at exploiting mitochondrial vulnerability in metabolically adaptable tumors. Full article

Background/Objectives: Vitex agnus-castus L. is a well-known medicinal herb shown to be effective in treating gynecological disorders. However, no systematic comparative studies have been conducted between V. agnus-castus leaf extract (VACLE) and Vitex agnus-castus seed extract (VACSE). We performed gas chromatography–mass spectrometry (GC–MS) analysis of VACLE and VACSE and measured total phenolic and flavonoid contents. Methods: The bioactivity testing included antioxidants, antibacterial, antifungal, anticancer, and antidiabetic activities. Results: A total of 55 GC–MS compounds were identified in VACLE and 34 in VACSE; isoamyl formate (27.96%) and a pyranone derivative (14.11%) were detected exclusively in VACLE, whereas cis-linoleic acid (40.58%) and palmitic acid (21.87%) predominated in VACSE. VACLE showed significantly higher TPC (94.12 vs. 54.12 mg GAE/g DW) and TFC (82.00 vs. 42.00 mg QE/g DW). The VACLE demonstrated moderate antioxidant activity and generally stronger bioactivity than VACSE, as evidenced by its lower ABTS⁺ radical scavenging IC₅₀ value (55 vs. 70 μg/mL), antibacterial activity (MIC: 6.25–50 vs. 12.5–100 μg/mL), anticancer activity against HepG2 cells (IC₅₀: 93.2 vs. 247.5 μg/mL), and antidiabetic activity through α-amylase inhibition (IC₅₀: 28.7 vs. 70.1 μg/mL). VACSE exhibited greater antifungal activity than VACLE against all tested Candida strains, with the highest activity observed against C. parapsilosis (MIC: 6.25 ± 2.26 μg/mL). VACLE induced transcriptional changes consistent with caspase-mediated apoptosis, characterized by increased expression of caspase-8, caspase-9, and Bax and decreased expression of Bcl-2/Bcl-xL, pending protein-level confirmation. Conclusions: In conclusion, VACLE exhibits notable antioxidant, antibacterial, anticancer, and antidiabetic properties, whereas VACSE shows greater antifungal activity. These findings highlight tissue-specific differences in phytochemical composition and in vitro biological activities and provide a basis for future studies involving compound isolation, mechanistic validation, toxicity assessment, and in vivo evaluation. Full article

The purpose of this study was to investigate the adverse effects of 1.5 μg·L⁻¹ environmentally relevant chlorantraniliprole (CAP) on oxidase biomarkers (juvenile, 2.5 g) for 2, 4, and 8 h and transcriptomic response (adult, 254.8 g) for 96 and 192 h in American shad Alosa sapidissima (Wilson, 1981). American shad is sensitive to pollutants and has become an important economic fish in China, especially for recirculating the aquaculture system and photovoltaic farming. For juvenile shad under short-time CAP exposure, acid phosphatase (ACP) and aryl hydrocarbon receptase (AHR) at the protein level significantly increased at 2 h, and for longer-time exposure, alkaline phosphatase (AKP), polyphenol oxidase enzyme (PPO), and tumor necrosis factor alpha (TNFα) at the protein level significantly decreased; ryanodine receptase (RYR) at the protein level was significantly increased at 8 h. Interestingly, malondialdehyde (MDA) contents, biomarkers of oxidative stress, were significantly decreased for depletion at 2 h and 4 h, while they increased for eliminating free radicals at 8 h via longer-time CAP exposure duration. With the same CAP exposure for adult shad, the number of congested and dilated sinuses of the liver changed, with fine granular brown pigmentation and vacuolization of hepatocytes at 96 h, while the sinuses and central veins were dilated and edematous degeneration occurred at 192 h for longer-time exposure. The detected enzymatic activities, except for adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK), significantly decreased, and MDA contents significantly increased in adult shad at 96 and 192 h. Ribosome, proteasome, spliceosome, protein processing in endoplasmic reticulum, oxidative phosphorylation, glycerophospholipid metabolism, biosynthesis of amino acids, ferroptosis, peroxisome, apoptosis, necroptosis, and mTOR signaling pathways were the most significantly enriched pathways. For qPCR verification, the genes ppa2, pla1a, psmb13a, pkz and stat1b were significantly upregulated, while hspa8b, capn2, tram2, asns, bcl2l1, diablo, and prkcb were downregulated in adult shad. The results reveal elevated oxidative stress causing time-dependent hepatic damage via 1.5 μg·L⁻¹ CAP exposure both in juvenile and adult shad. Full article