Aging is a long-term and complex process characterized by cellular changes, including cell cycle arrest, increased production of the senescence-associated secretory phenotype (SASP), and nuclear membrane disintegration. To accelerate the aging process, various animal models are made using chemical inducers, such as D-galactose (D-Gal). D-Gal has been extensively applied to induce aging in experimental animals; however, its effects on the gastrointestinal system, particularly the colon, remain underexplored. Observation of molecular aging markers will provide a valuable approach to assess these cellular changes. This study aimed to observe the expression of genes related to cell cycle arrest (p53 and Cdkn1a), SASP production (Il-6 and Il-1β), nuclear membrane disintegration (Lmnb1), as well as histological inflammation process in the colon of D-Gal-induced aging rat models. Twelve-week-old male Sprague-Dawley rats (n = 12) were divided into the control and D-Gal (100 mg/kg/day for 6 weeks) groups. The expression levels of aging-related genes (p53, Cdkn1a, Il-6, Il-1β, and Lmnb1) were assessed by reverse-transcription quantitative polymerase chain reaction. The D-Gal group exhibited inflammatory cell infiltration. Il-1β immunohistochemical score along with Il-1β expression was significantly elevated in D-Gal group (p < 0.01), suggesting SASP activation and pro-inflammatory signaling. No statistically significant differences were observed in the expression levels of p53, Il-6, Cdkn1a, or Lmnb1 expression. Molecular and histological results demonstrated that D-Gal administration induces colonic inflammation, characterized by increased Il-1β expression, which subsequently promotes potential cellular senescence.

Cancer persists as a major health burden, fueled not only by genetic mutations but also by profound epigenetic instability that rewires transcriptional programs and DNA repair networks. Among the most intensively studied epigenetic regulators are histone deacetylases (HDACs) and poly (ADP-ribose) polymerases (PARPs), whose dysregulation fosters genomic instability, unchecked proliferation, and therapeutic resistance. Pharmacological inhibition of HDACs or PARPs alone has achieved meaningful advances, yet intrinsic and acquired resistance, limited tumor selectivity, and relapse remain formidable barriers to durable efficacy. To address these challenges, attention has shifted toward rational combination strategies and, more recently, to the development of dual inhibitors. By integrating key pharmacophoric features from both HDAC and PARP inhibitors, these hybrids are designed to achieve balanced target engagement within a single scaffold, thereby maximizing synergy while reducing pharmacokinetic complexity. Mechanistically, dual blockade disrupts DNA repair fidelity, induces chromatin relaxation, and amplifies apoptotic signaling, thereby producing antitumor effects that exceed those of monotherapy. While this paradigm offers substantial promise, it is not without limitations, including potential off-target toxicity, challenges in optimizing linker chemistry, and the need for precise structure-activity relationship (SAR) refinement. This review consolidates structural, mechanistic, and SAR insights, emphasizing how dual HDAC/PARP inhibition represents a next-generation therapeutic strategy poised to overcome resistance and broaden the spectrum of effective cancer interventions.

Aging is a primary risk factor for disease progression in multiple sclerosis (MS). Because of this, treatments that can reduce the consequences of molecular aging, like senescence, have been proposed as a strategy to address disease progression. However, the effects of senolytics, a class of drugs which selectively ablate senescent cells, on the central nervous system are largely unknown. Here, we examined the effects of senolytic treatment on myelination and oligodendrocyte function in vivo using C57BL6/J mice and in vitro using primary rat oligodendrocyte cultures. Initial data showed that naïve young (3 to 4 mo) and aged (22 mo) C57BL6/J mice treated with dasatinib and quercetin (D+Q) developed significant demyelination compared to vehicle-treated controls, though no cell death was observed in the brain. In vitro, oligodendrocyte progenitor cells treated with D+Q in differentiation media exhibited significantly reduced myelin basic protein protein and morphological complexity, also without inducing cell death. Bulk RNA sequencing and ingenuity pathway analysis of D+Q treated oligodendrocytes identified differentially expressed genes associated with endoplasmic reticulum stress. These data suggest that D+Q evokes the unfolded protein response in oligodendrocytes, causing oligodendrocyte dysfunction and myelination failure. Due to the resemblance between oligodendrocytes treated with D+Q and those found in MS lesions, D+Q treatment offers a potential method to model an aspect of oligodendrocyte dysfunction relevant to MS. Therefore, understanding the mechanism by which D+Q perturb oligodendrocyte function may provide insight into some of the pathological features contributing to disease progression in MS.

Antimicrobial resistance (AMR) poses a serious threat to global public health. To tackle this challenge, identifying novel effective antibacterial targets and antibiotics is crucial. The serine kinase HPrK, widely present in gram-positive bacteria and mycoplasmas, primarily regulates carbon metabolism and virulence factor expression. In a previous study, we failed to knockout the hprK gene using the conventional homologous recombination method in the zoonotic Streptococcus suis. Here, we employed an anhydrotetracycline (ATc)-inducible promoter (AiP) and hprK fusion to replace its original expression cassette in the S. suis genome. However, in the absence of ATc, mutations in the AiP elements resulted in ATc-independent expression of HPrK, suggesting that HPrK is essential for S. suis and a potential antimicrobial target. Accordingly, a high-throughput inhibitor screening assay based on the kinase activity of HPrK was designed, and an inhibitor compound CDK9-IN-2 was identified. Molecular docking and bio-layer interferometry revealed that CDK9-IN-2 targets highly conserved residues D242, D249, and N281 on HPrK protein in several species of gram-positive bacteria. CDK9-IN-2 exhibited significant antibacterial effects against multidrug-resistant S. suis clinical isolates and various species of gram-positives and mycoplasmas with a minimum inhibitory concentration (MIC) of 16 to 32 μg/mL. Its therapeutic effect on S. suis infection was further proved in a Galleria mellonella larval infection model. Collectively, HPrK is a conserved kinase in gram-positives and mycoplasmas, and its essentiality is confirmed in S. suis. Based on this conserved target HPrK, an inhibitor with a broad antibacterial activity has been identified, suggesting HPrK is a promising antimicrobial target for antibacterial drug innovation.

Huntington's disease (HD) is a progressive neurodegenerative disorder caused by a CAG-repeat expansion in the Huntington gene (HTT). Herein, we describe the discovery of a series of HTT pre-mRNA-splicing modulators that promote the inclusion of a cryptic stop codon that in turn lowers levels of mutant Huntington protein (mHTT). Optimization of the starting thienopyridine amide core resulted in the discovery of the potent, CNS-penetrant, selective, and orally bioavailable HTT-splicing modulator BIO-6553. This lead compound is structurally distinct from existing splicing modulators, demonstrated significant HTT-lowering in both human cells and mouse YAC128 models, and has an attractive off-target profile from RASL- and RNA-seq analysis.

Glioblastomas (GBMs) are lethal brain tumors characterized by rapid growth and resistance to standard treatment. Resveratrol (RES) increases the levels of reactive oxygen species (ROS) and induces cell death in sensitive GBM cells. However, the death patterns induced by RES and their relevance to NFE2-related factor 2 (NRF2) remain unclear. The current study aimed to address these issues using RES-sensitive U251 and less-sensitive LN428 GBM cell lines, as well as an orthotopic GBM xenograft rat model. In silico analysis revealed high NRF2 expression in GBM tissues and a strong correlation with tumor progression in the TCGA dataset. After 48 h of 100 µM RES treatment, NRF2 levels remained stable in LN428 cells but significantly decreased by 2.3-fold in U251 cells, accompanied by suppressed growth and NRF2-regulated and ferroptosis-related xCT and GPX4 downregulation. Elevated Fe2+, ROS levels, lipid peroxidation, and ferroptotic frequency were evidenced in RES-treated U251 cells; meanwhile, apoptosis and reduced NRF2-HO-1 expression were also evident in U251 cells. Combined treatment with Ferrostatin-1 and Z-VAD FMK rescued 60% of U251 cells compared to RES-treated counterparts. In vivo, lumbar puncture (LP) administration of RES induced both ferroptosis and apoptosis in rat orthotopic GBM xenografts. These findings highlight the dual cell death induced by RES in sensitive GBM cells and identify NRF2 signaling status as a novel determinant of cellular response to RES treatment.

Intrahepatic cholangiocarcinoma (iCCA), a highly lethal subtype of liver cancer with increasing incidence and poor prognosis, displays profound molecular heterogeneity that limits therapeutic efficacy. Combining multi-omics with pathological subtyping in iCCA, we identified a strong association between the most aggressive molecular subtype and the large-duct pathological type, characterized by aberrant mucin overexpression and prominent myeloid cell infiltration. Through integrative analysis, MUC16 was identified as a key molecular marker specifically enriched in large-duct type intrahepatic cholangiocarcinoma (L-iCCA). Functional modulation of MUC16 markedly inhibited L-iCCA cell proliferation and migration. Furthermore, high-throughput drug screening identified the small-molecule compound OSMI-1 as a potent suppressor of MUC16 expression. Using primary iCCA cell lines and patient-derived organoids (PDOs), we confirmed that OSMI-1 inhibits L-iCCA cell proliferation and migration by downregulating MUC16. Mechanistically, OSMI-1 suppresses MUC16 expression by disrupting the transcriptional complex formed between OGT and IRF1. Employing a genetically engineered L-iCCA mouse model, we further validated the crucial role of MUC16 in tumor progression and neutrophil infiltration, demonstrated that OSMI-1 synergized with gemcitabine, effectively inhibiting L-iCCA growth. This study presents a pathological subtype-based precision therapeutic strategy for L-iCCA, thereby providing a foundation for novel translational approaches to the personalized management of this disease.

RNA-based therapeutics offer versatile strategies for disease prevention and treatment, yet precise control over gene expression remains a major challenge. Self-amplifying RNA (saRNA), derived from the alphavirus genome, could be engineered to contain multiple subgenomic promoters, providing a unique RNA-only architecture for modular and independent regulation of downstream genes. However, the potential of saRNA as a programmable gene circuit enabling small-molecule-controlled on/off regulation of gene expression has remained largely unexplored. In this study, we engineered a series of saRNA constructs incorporating multiple regulatory modules, including a destabilizing domain (DD), the RNA-binding protein L7Ae, a tetracycline-responsive repressor (TetR), and kink-turn (k-turn) RNA motifs. This design allows individual downstream genes driven by distinct subgenomic promoters to be independently and reversibly regulated by the FDA-approved small-molecule ligands trimethoprim (TMP) or doxycycline (Dox). The engineered saRNAs were encapsulated into lipid nanoparticles, and saRNA-mediated expression of luciferase or fluorescent reporter proteins was systematically evaluated both in vitro and in vivo. Our results demonstrate that TMP and Dox function as effective molecular switches to autonomously turn on or off specific gene expression programs encoded within a single saRNA molecule. Collectively, this work establishes saRNA as a programmable RNA gene circuit platform with ligand-responsive, multi-gene regulatory capability, providing a versatile foundation for the development of controllable RNA therapeutics and synthetic biology applications.

Glioblastoma (GBM) is a fast-growing primary brain tumor with high mortality and recurrence rates. Dysregulation of the cell cycle is a hallmark of GBM, and cyclin B1 (CCNB1) is a key regulator of the cell cycle. However, the role of CCNB1 in GBM remains unclear. In this study, we found that CCNB1 mRNA and protein expression levels were significantly higher in GBM tissues than normal tissues. High CCNB1 mRNA expression was associated with poorer prognosis in GBM patients. Single-cell and spatial transcriptomics data revealed that CCNB1+ cells represent a proliferative subcluster in GBM, annotated as proliferative cells, and characterized by the upregulation of cell cycle-related pathways. CCNB1 inhibition decreased the proliferation of GBM cells and impaired cell cycle progression from S phase to G2/M. Additionally, resveratrol could inhibit the expression of CCNB1 and its interacting gene polo-like kinase 1 (PLK1). Importantly, through in vitro and in vivo experiments, we found that resveratrol suppressed GBM cell growth with low toxicity. CCNB1 silencing combined with resveratrol treatment further inhibited the proliferation of GBM cells. Collectively, these data suggest that CCNB1 is highly expressed in GBM and may promote GBM progression. Inhibition of CCNB1 may represent a potential therapeutic strategy for GBM.

Natural products are a rich sources for developing anti-cancer drugs with low toxicity and high efficiency. Limonin has anti-cancer activity; however, its effect on cervical squamous cell carcinoma remains unreported. The aim of this study was to explore how Limonin affects ferroptosis in cervical squamous cell carcinoma (CESC) and its underlying mechanism. Based on differential gene analysis of the Gene Expression Omnibus database and drug target prediction of the Comparative Toxicogenomics Database, combined with molecular docking technology, potential anti-cancer targets of Limonin were identified. In vitro experiments were conducted to create epoxide hydrolase 2 (EPHX2) knockdown and overexpression cell lines. Relevant phenotypic experiments were conducted to verify how Limonin targeting EPHX2 affects cell proliferation and ferroptosis. Integrated bioinformatic analysis revealed EPHX2 as a key target of Limonin. Functional experiments showed that EPHX2 overexpression inhibited the proliferation of CESC and induced ferroptosis, while Limonin treatment could enhance EPHX2 expression in a concentration-dependent manner. Furthermore, EPHX2 knockdown could reverse the inhibitory effect of Limonin on CESC proliferation and alterations in ferroptosis-related indicators. This study results reveals a new mechanism by which Limonin induces ferroptosis in CESC by activating EPHX2, providing a new strategy for natural compound-based ferroptosis-targeted therapy.

Age-related macular degeneration (AMD) and diabetic retinopathy (DR) constitute leading causes of irreversible visual impairment; both are pathologically linked to chronic inflammation and endoplasmic reticulum (ER) stress in retinal pigment epithelial (RPE) cells. This study aimed to investigate the protective effects of hot-air-dried Eruca sativa Mill. extract (ESH) on lipopolysaccharide (LPS)- and thapsigargin (Tg)-induced inflammatory and ER stress responses, respectively, in ARPE-19 cells. ESH pretreatment significantly suppressed LPS-induced phosphorylation of nuclear factor kappa B (NF-κB), inhibitor of kappa B alpha, and c-Jun N-terminal kinase, indicating effective inhibition of inflammatory signaling cascades. At the transcriptional level, ESH markedly attenuated the expression of tumor necrosis factor-α mRNA, suggesting downstream prevention of NF-κB-mitogen-activated protein kinase-mediated inflammatory gene activation. Under ER stress conditions, ESH significantly attenuated the upregulation of CCAAT/enhancer-binding protein (C/EBP) homologous protein and X-box binding protein-1, along with reductions in the expressions of cleaved caspase-3 and -9, indicating mitigation of ER stress-associated retinal apoptosis. Additionally, ESH prevented Tg-inducible vascular endothelial growth factor (VEGF) mRNA expression, VEGF protein secretion, and intracellular calcium level. Strong positive correlations were observed between intracellular calcium and VEGF secretion (r = 0.888), and between VEGF mRNA and protein levels (r = 0.843), supporting a potential mechanistic link. Collectively, these findings demonstrate that ESH modulates inflammatory, ER stress, apoptotic, and angiogenic pathways, suggesting its potential as a functional dietary supplement to mitigate RPE dysfunction in AMD and DR.

Artificial control of gene expression in bacteria offers interesting prospects for influencing bacterial pathogenicity and antibiotic resistance. We show that the methyl-transferase cofactor, AdoHcy azide, can silence gene expression in modified plasmids in some strains of Escherichia coli, where ampicillin and kanamycin resistance as well as eGFP genes were selectively and independently disabled. The disabling of transcription is likely due to steric inhibition during transcription initiation, which is supported by Sanger and nanopore sequencing results. Both sequencing methods showed that 3-6 nucleotides were absent from around the modification site. Postgrowth, extracted AmpR/eGFP plasmid shows evidence of restriction, with sections of the plasmid, including the modification site, missing for the AdoHcy azide modified plasmids. Notably, the AdoHcy azide modification on the DNA appears to be resistant against demethylation in the BL21 strain of E. coli.

Aging is accompanied by chronic low-grade inflammation (inflammaging), driving age-related diseases. Urolithin A (UA), a gut microbial metabolite, possesses anti-inflammatory properties, yet its mechanism in hepatic aging remains unclear. This study investigated UA's effects on aging-associated inflammation and the involvement of Nur77 in D-galactose-induced macrophage senescence and mouse liver aging models using molecular docking, Western blotting, and immunoprecipitation. UA alleviated cellular senescence markers (p53, p21), suppressed pro-inflammatory factors (IL-6, IL-1β), and elevated anti-inflammatory IL-10. Mechanistically, UA enhanced Nur77 protein stability by inhibiting MDM2-mediated ubiquitination and degradation, thereby restoring inflammatory homeostasis. In vivo, UA ameliorated D-gal-induced liver injury and modulated the hepatic Nur77-MDM2 axis. Conclusion: UA stabilizes Nur77 by inhibiting its ubiquitination, alleviating hepatic aging-associated inflammation. This study identifies the MDM2-Nur77 axis as a potential therapeutic target for hepatic aging.

Chemotherapy-induced neutropenia (CIN) remains a major dose-limiting toxicity associated with myelosuppressive chemotherapy regimens. The development of therapeutic strategies capable of effectively restoring neutrophil production and function could address a critical unmet clinical issue. ZGSII, a bioactive compound derived from Sanguisorba officinalis, has shown potential in ameliorating leukopenia. To further evaluate its therapeutic applicability for CIN, a comprehensive understanding of its underlying mechanisms is essential. This study aims to assess the efficacy of ZGSII in mitigating cyclophosphamide-induced neutropenia and myelosuppression and to elucidate the underlying mechanism involved through transcriptome sequencing, protein-protein interaction network construction, and functional validation assays.

Nanoplastics (NPs) and salinity increasingly co-occur in agricultural systems. Here, we investigated how polystyrene NPs (502 nm, 10 mg L-1) impair rice (Oryza sativa L.) recovery from salt (50 mM NaCl). During stress, NPs synergistically amplified ionic toxicity, elevating Na+/K+ ratios 48% above additive predictions. Crucially, this synergism intensified to 60% during recovery, preventing homeostasis restoration. Transcriptomic analysis revealed that NP-salt interactions shifted from additive to antagonistic poststress, disrupting trehalose pathway regulation critical for osmotic adjustment. Additionally, coexposed plants failed to switch from stress- to growth-associated gene modules, exhibiting 34% fewer differentially expressed genes than salt-only plants. These findings demonstrate that NPs compromise transcriptional resilience by disrupting adaptive reprogramming, emphasizing the need for recovery-inclusive risk assessments.

Carotenoids serve as key quality indicators in yellow peaches, and enhancing their content is crucial for improving overall fruit quality. In this study, yellow peaches were treated with 1 mM abscisic acid (ABA) or 0.5 mM fluridone (Flu), an ABA biosynthesis inhibitor. ABA treatment upregulated the expression of key carotenoid biosynthesis genes, including Phytoene Synthase (PpPSY), Phytoene Desaturase (PpPDS), Zeta-Carotene Desaturase (PpZDS), β-Carotene Hydroxylase (PpCHYB), Lycopene β-Cyclase (PpLCYB), and Zeaxanthin Epoxidase (PpZEP), while downregulating the carotenoid degradation-related gene 9-Cis-Epoxycarotenoid Dioxygenase 1 (PpNCED1). This coordinated regulation promoted zeaxanthin, lutein, β-cryptoxanthin, and β-carotene, leading to a significant increase in total carotenoid content. In contrast, Flu treatment produced opposite effects, suppressing carotenoid biosynthesis and accumulation. The negative regulatory role of the transcription factor PpZAT10 in carotenoid accumulation was confirmed through transient overexpression and virus-induced gene silencing (VIGS) in peach fruits, as well as transgenic overexpression and CRISPR/Cas9-mediated knockout in peach callus. Furthermore, yeast one-hybrid (Y1H), electrophoretic mobility shift assay (EMSA), and dual-luciferase assays demonstrated that PpZAT10 directly binds to the promoter of PpPDS to repress its transcription and simultaneously activates the expression of PpNCED1 by binding to its promoter. These findings elucidate the molecular mechanism by which PpZAT10 mediates ABA-induced carotenoid accumulation in yellow peaches, providing a novel strategy for enhancing carotenoid content and, consequently, fruit quality.

Bortezomib, a first-in-class proteasome inhibitor, is widely used to treat multiple myeloma and other hematological malignancies. Despite its therapeutic efficacy, bortezomib causes peripheral neuropathy (PN) in approximately 20-30% of patients, often leading to dose reduction or discontinuation. Preventive or therapeutic approaches to bortezomib-induced PN are currently unavailable, as its precise mechanism remains unclear. In this study, we compared the effects of bortezomib and the second-generation proteasome inhibitor carfilzomib on peripheral nerve cells to identify candidate molecules involved in PN development. Transcriptome profiling of differentiated F11 cells, a hybridoma of a rat embryonic dorsal root ganglion and mouse neuroblastoma cell line N18TG2, revealed that bortezomib selectively upregulated α/β-hydrolase containing domain 4 (Abhd4), whereas carfilzomib did not. This finding was confirmed by quantitative RT-PCR and immunoblotting, which demonstrated consistent increases in Abhd4 mRNA and protein levels following bortezomib treatment. Functional analysis further revealed that Abhd4 overexpression promoted early apoptosis, suggesting a mechanistic link between bortezomib-induced Abhd4 elevation and neuronal vulnerability. Therefore, these results suggest that Abhd4 represents a candidate molecular signature associated with bortezomib-induced PN. Although further in vivo validation is needed, these findings warrant further investigation of Abhd4 as a potential contributor to bortezomib-induced PN.

As a globally cultivated leafy vegetable and an important economic crop, the production of Pakchoi (Brassica rapa ssp. chinensis var. communis) is increasingly threatened by high-temperature stress, often resulting in reduced yields and even complete loss. In this study, we systematically investigated the potential function of a proteoglycan, LEPS1, which was extracted from the spent substrate of Lentinula edodes, on regulating the thermotolerance of Pakchoi. We found that LEPS1 could significantly improve the thermotolerance of Pakchoi by modulating brassinosteroid (BR) signaling. Specifically, LEPS1 promotes BR biosynthesis and activates BR signal transduction; silencing of BR receptor BraBRI1 significantly downregulates LEPS1- and BR-induced thermotolerance in Pakchoi. We also found the core transcription factors BraBZR1 and BraBES1 directly binding to the promoters of heat-responsive gene and enhance their transcriptional activity, thereby improving the plant thermotolerance. Thus, our results revealed that the proteoglycan LEPS1 and BR enhances the thermotolerance of Pakchoi in a BraBRI1-BraBZR1/BraBES1 module dependent manner. These findings not only elucidated the molecular mechanisms through which the proteoglycan LEPS1 and BR induce thermotolerance, but also offer molecular targets and novel approaches to enhance the thermotolerance of Brassica crops.

Sodium nitroprusside alleviates drought in Lagenaria siceraria by regulating leaf wax synthesis, with LsCER1 pinpointed as a key drought-tolerance gene for breeding. Sodium nitroprusside (SNP) alleviates the drought stress of Lagenaria siceraria (L. siceraria); however, the underlying mechanisms remain poorly understood. In this investigation, gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling and transcriptomic analysis were employed to analyze the changes of different wax components and transcriptomes of 'Yayao' (L. siceraria) seedlings under control (CK), drought stress (DS), and drought + sodium nitroprusside (SP) treatments. Furthermore, correlation analysis was performed to integrate the two datasets. The results of GC-MS showed that exogenous sodium nitroprusside promoted the formation of cuticular wax, and the compositional analysis of cuticular wax revealed that fatty acids represented the most abundant constituent. KEGG pathway analysis indicated that differentially expressed genes (DEGs) in the three comparison groups were related to wax synthesis-related pathways such as ascorbic acid and aldehyde acid metabolism and galactose metabolism. A total of 116 DEGs were found in the three different comparison groups, of which 20 DEGs were related to the wax synthesis pathway. At the same time, four LsCER1 genes were identified, which were highly conserved and contained multiple hormone and abiotic stress response elements. The regulatory effect of L. siceraria cuticular wax synthesis by SNP has been provided and the foundation for future breeding strategies laid in the study.

Coffee silverskin (CSS), the major by-product of coffee roasting, is reported to contain bioactive compounds, including xanthines and polyphenols, showing promising potential for food and nutraceutical applications. This study investigated the beneficial effects of CSS hydroalcoholic extracts, which were chemically characterized by Attenuated Total Reflectance-Fourier-Transform Infrared Spectroscopy and ElectroSpray Ionization tandem Mass Spectrometry, on Caenorhabditis elegans physiology. CSS supplementation improved healthspan-related parameters and delayed aging-associated functional decline, without significantly extending lifespan in wild-type nematodes. Treated worms exhibited a 57% reduction in reactive oxygen species (ROS) levels and upregulation of antioxidant genes (gst-4 and sod-3), suggesting that CSS mitigates oxidative stress through the DAF-2/DAF-16 pathway. Under high-glucose diet conditions, CSS reduced lipid droplet accumulation and modulated the expression of metabolic genes, including upregulation of nhr-49 which is a key regulator of fatty acid oxidation. CSS restored lipid homeostasis and rescued the shortened lifespan of obese nhr-49 mutant worms, suggesting enhanced β-oxidation. Moreover, CSS modulated serotonergic signaling by increasing tph-1 and ser-6 expression, linking its effects to serotonin-mediated regulation of fat metabolism. Finally, CSS promoted the growth of probiotic strains, suggesting potential prebiotic properties. Overall, these findings identify CSS as a metabolic modulator capable of alleviating oxidative and metabolic stress, supporting its sustainable application in the development of functional foods and nutraceuticals.