Search Results (7,504)

Background/Objectives: Irisin is an exercise-associated myokine with reported effects on β-cell survival, insulin secretion, and pancreatic tissue protection in experimental models of metabolic stress. However, less is known about its influence on the developing endocrine pancreas under physiological conditions. Accordingly, the aim of the present study was to determine whether exogenous recombinant FNDC5/irisin is associated with changes in pancreatic endocrine-cell immunophenotype and islet-level markers of cell turnover in male and female Wistar rats. Methods: Thirty-two 3-week-old Wistar rats were assigned to control and irisin-treated groups, with males and females analyzed searately. Irisin was administered intraperitoneally at 100 µg/kg body weight once weekly for four weeks. Control animals received 0.9% NaCl. At the end of the experiment, serum cholesterol, fructosamine, and lipase were measured as supportive biochemical endpoints. Pancreatic sections were analyzed immunohistochemically for insulin, glucagon, somatostatin, PCNA, and cleaved caspase-3. Results: Irisin treatment was not associated with statistically significant changes in serum cholesterol, fructosamine, or lipase. In males, irisin increased insulin immunoreactive area, insulin-positive cell percentage, and insulin-positive cell density. In females, irisin decreased glucagon immunoreactive area and glucagon-positive cell density, whereas glucagon-positive cell percentage remained unchanged. Somatostatin immunoreactive area decreased after irisin administration in both sexes, while somatostatin-positive cell percentage and density increased in males and females. PCNA-positive cell percentage decreased in both sexes after irisin treatment. Islet cleaved caspase-3 immunoreactivity, assessed as optical density, was not significantly altered. Conclusions: Exogenous irisin modified pancreatic endocrine-cell immunophenotype in a sex- and cell-type-dependent manner in post-weaning rats. These changes occurred without significant alterations in serum lipase, fructosamine, or cholesterol and without a detectable increase in islet cleaved caspase-3 immunoreactivity under the applied analytical conditions. Full article

Background/Objectives: Although HSP70 and thioredoxin-1 (TRX1) are well known to counteract methamphetamine (METH)-induced neurodegeneration, no evidence on the effects of METH administration concerning the level, localization, and co-expression of HSP70 and/or TRX1 is available. Therefore, in the present study, carried out using methamphetamine-treated PC12 cells, we sought to analyze these effects along with the merging of specific markers for mitochondria, autophagy, lysosomes, and chaperone-mediated autophagy (CMA). Methods: The levels of specific proteins for specific cell compartments were assessed along with their merging. Thus, co-expression of HSP70 and TRX1 with lipidated LC3 (LC3-II), as well as lysosomal markers such as LAMP1 and LAMP2 (a specific marker for CMA), was assessed. Due to severe mitochondrial alterations produced by METH, both MitoTracker green and MitoTracker red were used in these experimental conditions. Light and electron microscopy, including in situ stoichiometry, were used in these experiments. Results: A moderate dose of METH, 100 μM, increases both TRX1 and HSP70; while HSP70 is preferentially co-expressed with the lysosome marker LAMP1, TRX1 is instead co-expressed with the marker LAMP2, which stains for CMA. Both chaperones increase along with cleaved caspase 3 and the autophagosome marker LC3-II. Conclusions: The outcome of the present study applies to METH-induced intoxication and is relevant in the fields of neurodegeneration, acute neuronal injury, and the domain of drug abuse. In fact, suppression of TRX1 enables METH-induced addiction, while overexpression of TRX1 or HSP70 produces the opposite effect. Full article

Esophageal squamous cell carcinoma frequently develops through a prolonged precancerous phase, creating an opportunity for intervention. Zishui Qinggan Yin (ZSQGY) is a multi-component botanical preparation, but its effects in esophageal precancer and its formulation-level chemical profile remain incompletely defined. We monitored a predefined 40-analyte panel by UPLC-Q Exactive/MS in three independently extracted aliquots from the same formulation batch administered to mice. A 4-nitroquinoline-1-oxide model was then evaluated by consensus histopathology, mouse-grouped whole-slide digital pathology, network-based target prioritization, molecular docking, exploratory drug affinity responsive target stability (DARTS), Western blotting, qPCR, and serial-section immunohistochemistry. Signals corresponding to 38 predefined analytes were reported. ZSQGY treatment was associated with a lower-grade lesion pattern in the evaluated mouse-level specimens. Exploratory DARTS was analyzed in two independent n = 6 tissue-background datasets. In the original 4-NQO model-background cohort, ex vivo ZSQGY (100 μg/mL) increased the AKT1-immunoreactive relative-protection signal in five of six lysates (mean 2.04 ± 1.15). In an additional cohort from mice that had received ZSQGY in vivo, ex vivo ZSQGY produced mean relative-protection values of 1.21, 1.75, and 1.92 at 50, 100, and 200 μg/mL, respectively. The nominal exact p value for the 100 μg/mL versus vehicle contrast in the additional exploratory dataset was 0.0313; This exploratory result should be interpreted cautiously. Neither dataset establishes direct binding or in vivo target occupancy. PI3K p110α DARTS was technically inconclusive. Endpoint tissues showed lower AKT, mTOR, S6K1, and 4E-BP1 phosphorylation, together with PTEN-associated changes, reduced Ki-67, increased cleaved caspase-3 staining, and selected downstream transcript changes, including a non-significant downward trend in PARP1. These findings support a preclinical association between ZSQGY treatment, attenuation of esophageal precancer progression, and AKT-centered PI3K/AKT/mTOR pathway modulation. They do not establish direct binding, in vivo target occupancy, pathway necessity, active constituents, or causal mediation. Full article

Background: Glucose handling in hepatocyte-derived cells integrates insulin-responsive AKT signaling with GLUT1-mediated glucose transport, jointly regulating substrate trafficking, glycogen synthesis, and cell survival. MK-2206, a potent allosteric AKT inhibitor, and BAY-876, a highly selective GLUT1 inhibitor, represent targeted approaches to disrupt these processes. However, the consequences of simultaneous inhibition of both nodes in hepatocyte-derived cells remain incompletely understood. Objective: This study aims to evaluate the impact of simultaneous AKT inhibition and GLUT1 blockade on target engagement, cell viability, proliferation, cell death, glucose uptake, and glycogen storage in HepG2 cells. Methods: HepG2 cells were treated with MK-2206, BAY-876, or their combination under basal hyperglycemic and acute insulin-challenged conditions. Multi-model synergy, AKT/GLUT1 pharmacological effects, cell viability, proliferation, cell death, glucose uptake, and glycogen storage were assessed, alongside exploratory molecular docking analyses of both inhibitors against their respective targets. Results: Dual AKT and GLUT1 inhibition produced combination-specific reductions in cell viability and showed a pattern of concurrent antiproliferative effects and increased Annexin V/PI positivity without detectable caspase-3 activation under insulin-challenged conditions. Furthermore, the dual inhibition of AKT signaling and GLUT1-mediated glucose uptake produced combination-specific reductions in basal glycogen content, while glucose uptake reductions showed AKT-dominant patterns without significant combination superiority over MK-2206 alone under either metabolic state. Exploratory molecular docking of MK-2206 and BAY-876 against AKT1 and SLC2A1/GLUT1, respectively, generated computationally predicted binding poses consistent with the established binding modes of these inhibitors. Conclusions: Dual AKT/GLUT1 targeting produces endpoint-specific patterns of pharmacological interaction on insulin-responsive metabolism, proliferation, and cell death in HepG2 hepatocyte-derived cells, distinguishing combination-specific effects from AKT-driven effects. These findings support an endpoint-specific pharmacological framework in which AKT signaling and BAY-876-mediated GLUT1 pharmacological effects jointly influence glucose handling and cell fate in a hepatocyte-derived model. Full article

Sulfometuron-methyl (SM), as a sulfonylurea herbicide, is widely used in agricultural production. In recent years, it has also been employed in the control and management of Spartina alterniflora in some coastal areas of China. Its potential risks to aquatic ecosystems have drawn increasing attention. However, the current data on the systematic toxicity assessment of SM in model organisms is still very limited. This study used zebrafish (Danio rerio) as an in vivo model to systematically evaluate the toxic effects of SM on early embryonic development and focused on the mediating mechanism of the Wnt/β-catenin signaling pathway in it. Zebrafish embryos were exposed to different concentrations (10, 20, 40 mg/L) of SM starting from 1.75 hpf until 72 hpf. The results showed that SM significantly increased the embryo mortality rate, decreased the hatching rate, and induced developmental malformations such as shortened body length and yolk sac edema in a concentration-dependent manner. At the mechanism level, SM exposure significantly reduced the number of H3P-positive mitotic active cells in the embryos, suggesting that fewer cells entered mitosis; meanwhile, the activities of superoxide dismutase (SOD) and catalase (CAT) showed compensatory changes, indicating that oxidative stress was induced in the embryos, which may contribute to subsequent cellular damage. qRT-PCR analysis further revealed that SM exposure downregulated the transcriptional levels of cell cycle-related genes (CyclinD1, CDK4, CDK6), while upregulating the expression of apoptosis-related genes (p53, bax, caspase-9, caspase-3), suggesting that cell cycle arrest and the dysregulation of the endogenous apoptotic pathway were activated. Additionally, SM exposure could dysregulate the Wnt/β-catenin signaling pathway, specifically manifested as significant upregulation of target genes C-myc and Ctnnb2, and significant downregulation of the negative regulatory factor Axin2. Using the Wnt/β-catenin pathway-specific inhibitor IWR-1 for intervention could partially reverse the embryonic developmental retardation phenotypes induced by SM, providing additional evidence for the mediating role of this pathway in the developmental toxicity of SM. In summary, this study systematically elucidates the multi-level molecular mechanism by which SM induces oxidative stress and cell apoptosis through the abnormal dysregulation of the Wnt/β-catenin signaling pathway, reduced mitotic activity, and ultimately drives the toxicity of zebrafish embryo development. This provides new experimental evidence and a theoretical basis for the aquatic ecological risk assessment of SM. Full article

Cadmium (Cd) is a ubiquitous environmental pollutant with an exceptionally long biological half-life (10–30 years) and high multi-organ toxicity. While chronic cadmium exposure has been extensively studied, the pathological mechanisms and prophylactic strategies for acute cadmium poisoning remain poorly defined. Here, we established an acute cadmium intoxication model in C57BL/6 mice via intraperitoneal injection of CdCl₂ (5 mg/kg) and assessed pathological and molecular changes 12 h post-exposure. Histopathological examination revealed overt hepatic damage and mild extrahepatic changes in kidney and spleen. Mechanistically, acute cadmium challenge disrupted systemic redox homeostasis, as evidenced by significant reductions in superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and total antioxidant capacity (T-AOC). Concurrently, cadmium triggered a robust inflammatory response, upregulating pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and lactate dehydrogenase (LDH) release, and activated the pyroptotic pathway, as shown by elevated cleaved caspase-1 and GSDMD-N levels. To identify potential prophylactic agents, we investigated nifuroxazide (NFX), a multifunctional agent with known anti-cancer, antioxidant, and anti-inflammatory properties. Our results indicate that NFX pretreatment protects against cadmium-induced acute liver injury. Mechanistically, these data support the involvement of the USP21–AIM2 axis in this protection. USP21, a deubiquitinase, stabilizes AIM2 by removing its ubiquitin chains. Cadmium exposure increased USP21-mediated AIM2 deubiquitination, decreased AIM2 ubiquitination, and promoted AIM2 protein accumulation, accompanied by AIM2 inflammasome activation and hepatic stellate cell pyroptosis. In contrast, NFX pretreatment was associated with reduced USP21-mediated AIM2 deubiquitination, increased AIM2 ubiquitination and reduced AIM2 protein accumulation, and suppressed hepatic stellate cell pyroptosis. Collectively, these findings indicate that NFX pretreatment protects against acute CdCl₂-induced hepatic injury with mild improvements in extrahepatic tissue histology, and support the involvement of the USP21–AIM2 axis in regulating AIM2 inflammasome protein stability. Full article

This article describes the synthesis, structure confirmation, and evaluation of physico-chemical, pharmacological, and pharmacokinetic properties of new para-fluorophenylated dihydroimidazotriazinones. Based on the strategy of heteroannulation, and the concept of fluorine–hydrogen isosterism, an efficient and simple synthetic route was developed. The target heterobicycles (10–18) were obtained by the reaction of nucleophilic building blocks, i.e., 1-(R-phenyl)-2-hydrazinylideneimidazolidine hydroiodides (1–9), with an electrophilic two-carbon synthon, i.e., 2-(4-fluorophenyl)-2-oxoacetic acid. The structures of the new compounds were confirmed by spectroscopic data. The spectrum of anticancer activity, selectivity profile, and effect on caspase levels, as well as the haemolytic and antihaemolytic properties of the molecules were assessed in in vitro and ex vivo studies. Most compounds exhibited antiproliferative activity against human solid tumour and leukaemic cells (superior/comparable to anticancer drugs) with low toxicity to normal cells. Moreover, they revealed a broader spectrum of anticancer activity than previously obtained isosteres without fluorine substitution, suggesting that the isosteric replacement of hydrogen with fluorine was a fruitful modification. The most selective molecules were able to increase the levels of apoptotic caspases in lung, cervical, and breast cancer cells. Molecular docking results revealed that although all fluorophenylated fused triazinones exhibited affinity for the adenosine A_2A and A_2B receptors, ligands 13, 16, and 17 emerged as promising dual-target candidates due to their superior interaction profiles and stronger binding affinities for both receptor subtypes. All the compounds proved to be safe for red blood cells, as they did not induce any haemolytic effects. Additionally, most of them revealed protective effects on oxidatively stressed erythrocytes, and their antihaemolytic activity was better or comparable to that of antioxidant standards. In silico ADME (absorption, distribution, metabolism, excretion) profiling demonstrated favourable drug-like and pharmacokinetic properties of compounds. Lipophilicity of the molecules was measured by RP-HPLC on an ODS-2 column using an appropriately selected mobile phase. The retention factors, logs k, for most compounds showed a strong correlation with both in silico log P (logarithmic n-octanol/water partition coefficient) and log P_HSA (logarithmic human serum albumin/water partition coefficient) values. In summary, among the nine original fluorophenylated fused 1,2,4-triazinones studied, five molecules (11, 13, 14, 15, and 17) seem to be the most promising anticancer drug candidates suitable for further development. Full article

Due to the hypoxic environment caused by global warming and intensive aquaculture process, fish are susceptible to oxidative damage and death. Eucommia ulmoides extract (EUE) has demonstrated antioxidant and anti-inflammatory properties. However, whether EUE can mitigate hypoxia-induced negative effects remains uninvestigated in fish. This study investigated the effect of 0 and 1.0 g/kg EUE on acute hypoxia-induced damage in largemouth bass and its potential mechanisms. The results showed that EUE alleviated the deterioration of plasma activities (ALT and AST) induced by acute hypoxic stress, decreased the number of macrophages and increased the total number of blood cells. EUE was effective in reducing the level of apoptosis and the content of ROS, Ca²⁺, and NO in blood cells. Additionally, EUE alleviated liver tissue injury caused by acute hypoxic stress, which could be associated with increased total antioxidant capacity and antioxidant enzyme activities (SOD, CAT and GSH-Px). Furthermore, EUE up-regulated the expression levels of Nrf2 and CAT, activated XBP1 and IRE expression and down-regulated GRP78 expression, relieving the endoplasmic reticulum (ER) stress response. The down-regulation of Caspase3 and Caspase9 is evidence that EUE inhibited apoptosis in acute hypoxic stress. In summary, EUE reduced oxidative damage, ER stress, and apoptosis caused by hypoxic stress via the Nrf2/Keap1 signaling pathway in largemouth bass. This study presents a part of the theoretical foundation for the EUE to resist damage caused by hypoxic stress, thereby establishing a basis for sustainable development in aquaculture. Full article

Caprine kobuvirus (CKoV) is a member of the Picornaviridae family that causes enteritis in goats and poses a significant health threat, yet its molecular pathogenesis remains largely unknown. Here, we show that experimental infection of goats with CKoV induces apoptosis in the spleen, lungs, and intestine tissues. In vitro, CKoV infection of HEK293T cells triggers apoptosis, with the viral structural protein VP1 identified as sufficient to induce apoptosis. VP1 activates the mitochondrial (intrinsic) apoptotic pathway, as evidenced by increased reactive oxygen species (ROS) production, activation of caspase-9 and caspase-3, elevated Bax/Bcl-2 ratio, cytochrome c release, and Bax translocation to mitochondria. Notably, VP1 associates with the pro-apoptotic protein BAD (BCL2-associated agonist of cell death) and co-localizes with it in mitochondria. Overexpression of BAD enhances VP1-induced apoptosis, whereas BAD knockdown by siRNA suppresses it, demonstrating that BAD plays a contributory role in VP1-induced apoptosis. These findings uncover a novel VP1–BAD mitochondrial apoptosis axis in CKoV pathogenesis and suggest that VP1 may merit further investigation as a potential molecular target for antiviral intervention against CKoV-associated enteritis in goats. Full article

Background and Objectives: Drug repurposing provides an opportunity to identify anticancer activities among established pharmacological agents. Labetalol (LB), an α₁- and non-selective β-adrenergic receptor antagonist, has been insufficiently investigated in melanoma. This study evaluated the potential anti-melanoma activity of LB in A375 human melanoma cells, compared its effects with those observed in HaCaT immortalized non-tumoral keratinocytes, and investigated the cellular mechanisms associated with LB-induced cytotoxicity. Materials and Methods: A375 and HaCaT cells were exposed to 75–500 μM LB for 24 h. Cell viability and lysosomal dye retention were assessed using MTT and NRU assays. A DMSO-only concentration series (0.075–0.50% v/v) was additionally evaluated by MTT in both cell lines. Mitochondrial membrane potential, mitochondrial staining patterns, nuclear morphology, cytoskeletal organization, caspase-3/7 and caspase-9 activities, and plasma membrane integrity were evaluated using JC-1, MitoTracker Red CMXRos, immunofluorescence, luminescence-based caspase assays, and AO/PI staining. The acute irritation potential of 500 μM LB was assessed using the HET-CAM assay. Results: DMSO alone did not significantly reduce viability over the investigated concentration range in either cell line. LB reduced A375 cell viability in a concentration-dependent manner, with a 24 h IC₅₀ of 422.5 μM and a viability of 33.61% at 500 μM. Under the same experimental conditions, HaCaT viability remained at 89.01% at 500 μM, and an IC₅₀ was not reached within the investigated concentration range. At the highest concentration, neutral red uptake and the JC-1 aggregate/monomer ratio decreased to approximately 33% and 24% of the corresponding control values, respectively. The apoptotic index increased from approximately 3% in control cells to 31%, while caspase-3/7 and caspase-9 activities increased to approximately 482% and 324% of the control. Qualitative imaging demonstrated mitochondrial staining redistribution, cytoskeletal disorganization, apoptosis-associated morphology, and loss of plasma membrane integrity at higher concentrations. In the HET-CAM assay, 500 μM LB produced a mean irritation score of 0.71 ± 0.27, within the non-irritant range. Conclusions: LB displayed concentration-dependent in vitro cytotoxicity in A375 melanoma cells, whereas HaCaT keratinocytes showed limited changes under the investigated conditions. LB treatment was also associated with mitochondria-associated apoptotic signaling in A375 cells. These findings provide preliminary evidence supporting further investigation of LB within a melanoma-directed drug-repurposing strategy. Additional studies are required to establish receptor dependence, achievable local exposure, safety, and translational relevance. Full article

Crystal-induced arthropathies, including gout and calcium pyrophosphate deposition disease, are characterized by activation of inflammatory and oxidative pathways involving nuclear factor-kappa B (NF-κB) signaling, inflammasome activation, and reactive oxygen species (ROS) generation. This study evaluated the anti-inflammatory and antioxidant effects of 2,4,5-trimethoxy-2′-trifluoromethylchalcone (CH) in experimental models of crystal-induced inflammation and gouty arthritis. CH was evaluated in lipopolysaccharide (LPS)/adenosine 5′-triphosphate (ATP)-stimulated murine peritoneal macrophages and in murine models of CPPD crystal-induced air pouch inflammation and monosodium urate (MSU)-induced gouty arthritis. Cytokine production was determined by ELISA, whereas NF-κB and caspase-1 activation were evaluated by Western blot. ROS generation and nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) were also assessed. CH reduced interleukin (IL)-1β, IL-18, tumor necrosis factor (TNF)-α, and IL-6 production in activated macrophages and attenuated lactate dehydrogenase release associated with pyroptosis. Mechanistically, CH reduced p65 NF-κB phosphorylation, caspase-1 activation, and ROS generation while promoting Nrf2 nuclear translocation. In vivo, CH reduced leukocyte infiltration, myeloperoxidase activity, inflammatory cytokine levels, and paw edema in CPPD- and MSU-induced models. These findings suggest that CH acts as a multitarget compound capable of simultaneously modulating inflammatory and oxidative pathways involved in crystal-induced arthropathies, which may represent a potential advantage over approaches targeting a single inflammatory pathway. Full article

Familial Alzheimer’s disease (FAD) is an accelerated form of dementia affecting cholinergic neurons. Despite several efforts, no single drug or treatment has demonstrated complete efficacy. Therefore, finding effective therapeutic agents is imperative. Previous studies have shown that the PSEN1 I416T variant induces FAD-like neuropathology in cholinergic-like neurons (ChLNs), characterized by the intracellular accumulation of the Aβ (iAβ) peptide, the oxidation of the stress sensor protein DJ-1, the abnormal phosphorylation of the tau protein at serine 202/threonine 205 (pS202/T205), the loss of mitochondrial membrane potential (ΔΨ_m), and activation of the pro-apoptotic proteins tumor protein 53 (TP53), Jun proto-oncogene, AP-1 transcription factor subunit (c-JUN), p53 upregulated modulator of apoptosis (PUMA), and cleaved caspase-3 (CC3). We report for the first time that PSEN1 I416T induces abnormal phosphorylation of Leucine-rich repeat kinase 2 (LRRK2) kinase at residue serine 935 (S935), concomitant with phosphorylated alpha-synuclein (αSYN) at residue serine 129 (S129) and abnormal accumulation of autophagosomes and atypical increase in vesicular lysosomal pH, thereby provoking a profound alteration in autophagy in ChLNs. Here, we also demonstrate for the first time that the potent LRRK2 inhibitor PF-06447475 (hereafter referred to as PF475) almost completely attenuated PSEN1 I416T-induced proteinopathy, oxidative stress (OS), and apoptosis and improve autophagy to an activity comparable to untreated wild-type (WT) ChLNs. Overall, PF475 restored the survival of mutant ChLNs. Taken together, these findings suggest that PF475 is an excellent pharmacological tool with which to investigate the regulation of LRRK2-associated pathological signaling in the PSEN1 I416T familial Alzheimer’s disease (FAD) model. Full article

Background/Objectives: Diffuse midline gliomas (DMGs), including diffuse intrinsic pontine glioma (DIPG), present a significant medical challenge due to their aggressive nature and poor prognosis. Widespread overexpression of the interleukin-13 receptor subunit alpha-2 (IL-13Rα2) across gliomas makes it a promising therapeutic target. Method: Using epitope mapping through a molecular imprinting (EMMI) technique, we developed nanosized molecularly imprinted polymers (nanoMIPs) that specifically recognize different IL-13Rα2 domains. Results: Functional assays showed that nanoMIPs binding to the surface of IL-13Rα2 reduced glioma cell viability, while sparing non-tumor cells. Consistent with receptor-mediated interactions, these nanoMIPs were selectively accumulated onto glioma cells, a specificity validated using a reverse-epitope nanoMIP negative control (nanoMIP-1*). IL-13Rα2-specific nanoMIPs suppressed phosphorylation of ERK1/2, AKT, and STAT3, disrupting major survival pathways. Moreover, these nanoMIPs activated the pro-apoptotic caspase-3/7 pathway, confirming apoptosis induction. Conclusions: Our findings demonstrate that IL-13Rα2-specific nanoMIPs, without a pharmacological payload, exert strong anti-proliferative and pro-apoptotic effects on DMG cells. Collectively, nanoMIPs represent a robust, reproducible synthetic antibody nanoplatform for targeted therapies in cancer and other diseases characterized by cell-surface protein dysregulation. Full article

Cisplatin resistance remains a significant challenge in the effective treatment of breast cancer, underscoring the need for novel combination-based therapeutic strategies. Background: This study investigated the anticancer effects of ellagic acid (EA) and metformin (MET), both individually and in combination, against parental and cisplatin-resistant breast cancer models. To our knowledge, this is the first study to evaluate the EA-MET combination in the context of cisplatin resistance in breast cancer. Methods: The in vitro antiproliferative and apoptotic effects of EA and MET were assessed using MTT and caspase-3 assays in parental (EMT-6/P) and cisplatin-resistant (EMT-6/CPR) cells. An in vivo study was also conducted to evaluate the anti-tumor effects of the treatment combinations in mice bearing EMT-6/P or EMT-6/CPR tumors. Liver enzymes and creatinine levels were measured to assess potential hepatic and renal toxicity. Results: In vitro combination index analysis indicated a synergistic interaction between EA and MET in EMT-6/P cells (CI = 0.45) and a strong synergistic interaction in EMT-6/CPR cells (CI = 0.29) under the tested conditions. In vivo, the addition of the combination to cisplatin resulted in significant tumor size reductions of 73.83% and 56.25% in EMT-6/P and EMT-6/CPR bearing mice, respectively, along with tumor regressions of 66.6% and 50%, respectively. Conclusions: Thecombination of metformin and ellagic acid shows promise as a potential treatment for cisplatin-resistant breast cancer. Further research is needed to elucidate the precise anticancer mechanisms underlying this combination. Full article

Inflammatory bowel disease is characterized by recurrent mucosal inflammation, epithelial barrier disruption, and dysregulated cell death, highlighting the need for safe, multi-target interventions. Natural astaxanthin, renewably sourced from Haematococcus pluvialis or Phaffia rhodozyma, has shown greater tissue accumulation or antioxidant activity than synthetic astaxanthin in some preparations. Based on the complementary antioxidant, anti-inflammatory, and intestinal barrier-protective activities of its individual components, the astaxanthin–turmeric extract–honeysuckle extract combination (ATH) was investigated for its protective effects in a mouse model of dextran sulfate sodium (DSS)-induced colitis. Thirty-two male C57BL/6J mice were randomly allocated to four cohorts: Control, DSS, DSS + ATH, and ATH. ATH intervention mitigated the DSS-associated decline in body mass, lowered clinical disease scores, attenuated DSS-induced colon shortening, and ameliorated mucosal lesions. In DSS-treated mice, ATH attenuated goblet-cell loss, reduced intestinal permeability, and restored ZO-1 and occludin expression. ATH treatment also reduced DSS-induced increases in colonic IL-1β, IL-6, TNF-α, IL-18, and malondialdehyde levels while improving superoxide dismutase activity and glutathione content. At the signaling level, ATH reduced phosphorylated NF-κB and NLRP3 immunoreactivity and decreased N-GSDMD accumulation within E-cadherin-positive epithelial regions. Western blotting further showed reduced NLRP3 expression, caspase-1 cleavage, and N-GSDMD formation, although the p-NF-κB/NF-κB ratio was not significantly changed. Collectively, ATH alleviated DSS-induced colitis by improving mucosal barrier and redox–inflammatory homeostasis while attenuating the NLRP3/caspase-1/GSDMD pyroptotic cascade. These findings support the further development of ATH as a multi-component natural intervention for intestinal inflammatory injury. Full article