Capecitabine (CAP), a chemotherapy prodrug, is associated with a significant dose-limiting hepatotoxicity. This study evaluated the protective efficacy of melatonin (MEL) and cerium oxide nanoparticles (CeO₂ NPs) against CAP-induced toxicity. Fifty male Wistar rats were divided into five groups: control, CAP-only (500 mg/kg/week), CAP + MEL (10 mg/kg/day), CAP + CeO₂ NPs (30 mg/kg/day), and a combination therapy group (CAP + MEL + CeO₂ NPs). After a six-week treatment period, assessments included alanine and aspartate transaminases (ALT, AST); hepatic oxidative stress markers [glutathione peroxidase-1 (GPx-1), superoxide dismutase (SOD), and malondialdehyde (MDA)]; apoptotic markers [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (BAX) mRNA, cleaved caspase-3 (c-caspase-3), and protein 53 (p53)]; pro-survival signaling [extracellular signal-regulated kinase (ERK1/2)]; and histopathological evaluation of liver tissue. CAP administration induced significant liver damage, characterized by elevated ALT and AST, depletion of GPx-1 and SOD, increased MDA, and activation of the mitochondrial apoptotic pathway, indicated by an elevated BAX/Bcl-2 ratio and increased levels of p53 and c-caspase-3. This was accompanied by suppression of ERK1/2 signaling and severe histopathological alterations. Both MEL and CeO₂ NPs, significantly attenuated these deleterious effects. The combination therapy demonstrated a comparatively superior effect, resulting in a near-complete normalization of all measured parameters, including oxidative stress, apoptosis, ERK1/2 activity, and liver histoarchitecture. Melatonin and CeO₂ NPs provide hepatoprotection via distinct yet complementary mechanisms. Their combined administration presents a superior strategy to mitigate CAP-induced liver injury by concurrently counteracting oxidative stress, inhibiting apoptosis, and enhancing pro-survival signaling.
Acridine derivatives exhibit diverse biological activities, yet their potential to modulate angiogenesis remains underexplored. This study evaluated 9-phenyl acridine (ACPH) as a small-molecule ligand capable of engaging vascular endothelial growth factor (VEGF) and altering conformational features relevant to receptor recognition, with the working hypothesis that ACPH may modulate VEGF structural dynamics in regions involved in VEGF-VEGFR interaction rather than directly inhibiting receptor binding. Structure-based binding was assessed using AutoDock, PatchDock, and GOLD, with VEGF as the molecular target. Complex stability and conformational effects were examined by 50-ns molecular dynamics simulations, including RMSD, residue-level flexibility, compactness, solvent exposure, and interaction energetics. Functional activity was evaluated using the chick embryo chorioallantoic membrane (CAM) assay at 0.04 nM, 0.4 nM, and 0.8 nM, with ImageJ-based quantification of vascular features over 24–48 h. Docking consistently positioned ACPH within a VEGF pocket, with predicted contacts involving Phe36, Ile46, Phe47, and Lys48 (chain A) and Cys60, Cys61, Asn62, Asp63, Glu64, and Glu67 (chain B). Across simulations, ACPH association was associated with reduced conformational variability of VEGF, particularly in regions relevant to protein-protein interaction, supported by an estimated ligand-protein interaction energy of approximately − 26.77 kcal/mol derived from trajectory-based and interpreted qualitatively rather than as a rigorous thermodynamic binding free energy. In ovo, ACPH exposure was associated with a dose-dependent reduction in CAM vascularization, with observable effects at 0.4 nM and more pronounced effects at 0.8 nM; however, this represents a phenotypic observation and does not establish VEGF-specific inhibition or a defined anti-angiogenic mechanism. Prolonged exposure at 0.8 nM was also associated with reduced embryo viability, indicating that non-specific toxicity may contribute at the highest dose. Docking and MD simulations consistently positioned ACPH within a transient VEGF cavity and were associated with reduced conformational drift and residue-level fluctuations. In the CAM model, ACPH exposure was associated with dose-dependent vascular reduction, with reduced embryo viability at 0.8 nM suggesting possible non-specific toxicity; these findings warrant receptor-facing validation and cytotoxicity-controlled endothelial assays.
Diabetes is associated with a number of significant long-term effects. In this study we consider that purslane which possesses numerous of pharmacological properties, and metformin, an antidiabetic drug, may have a therapeutic effects on diabetes-induced memory impairments in rats. Forty male albino rats were randomly divided into five groups. Group I served as control group. The other four groups were first fed on HFD followed by a single interpretonial (i.p.) dose of STZ at a dose of (35) mg/kg then the groups were divided as following Group II diabetic group Group III, PEE group administered with oral dose of purslane ethanolic extract (100 mg/kg) for another four weeks. Group IV, MET group administered with oral dose of metformin (100 mg/kg) for another four weeks. Group V (PEE + MET) administered with oral dose of combination of both purslane ethanolic extract (50 mg/kg) and MET (50 mg/kg) for another four weeks. During the treatment rats were tested for memory and learning abilities (Morri’s water maze test). Hippocampal samples were collected for biochemical, and histological measurements. Biochemical evaluation included (NO and TBARS) as an oxidative stress marker, (GSH, GPX, SOD, Catalase) as antioxidant, and inflammatory cytokines (tumor necrosis factor-α and interleukin-1β, interleukin-IL-6). Also, P-tau protein, (dopamine and GABA) as neurotransmitters, and for cholinergic system (acetylcholinesterase) were assessed, in addition to histological examinations of hippocampus. Diabetic rats showed a marked cognitive impairment in the Morris water maze test and alteration in the other biochemical and histological features. Intrestingly, PEE and MET treatments partially dramatically enhanced antioxidant levels. Also, reduced oxidative stress, pro-inflammatory mediators, and, phosphorylated tau levels. In addition, PEE and MET treatments partially modulated neurochemical profiles associated with memory function. The combined PEE + MET treatment showed the most pronounced improvement, reflecting synergistic effects. Individual data points highlighted consistent trends across animals. Also, it exhibited a significant restoration of normal hippocampal architecture, as confirmed by hematoxylin and eosin staining. The data obtained indicated that PEE, either alone or in combination with MET, has strong neuroprotective potential against STZ/HFD-induced diabetes. These safeguarding effects are probably because of its strong anti-inflammatory and antioxidant properties.
Abstract Background Acetyl tributyl citrate (ATBC), a widely used plasticizer, has raised health concerns due to its potential environmental persistence and human exposure, but its toxicological effects on sarcoma remain unclear. Methods We employed an integrated approach combining network toxicology, molecular docking, survival analysis, and experimental validation to systematically investigate ATBC’s impact on sarcoma. Results Multi-database screening identified 102 overlapping targets. Protein–protein interaction analysis revealed six hub genes: TLR4, ESR1, PPARG, SIRT1, NFKB1, and PTGS2. Functional enrichment analysis indicated significant involvement in cancer-related pathways, including PI3K–Akt, HIF‑1, and AGE–RAGE signaling. Molecular docking predicted potential interactions between ATBC and these targets, with binding energies ranging from -1.15 to -2.99 kcal/mol. Survival analysis associated high PPARG expression with poor prognosis, while high TLR4 and ESR1 expression correlated with better survival. In vitro experiments showed ATBC promoted proliferation and migration of sarcoma cells. qPCR results confirmed ATBC downregulated TLR4 and ESR1 and upregulated PPARG, aligning with clinical prognostic trends. Conclusions These findings suggest ATBC may exert tumor-promoting effects by modulating core targets and pathways, highlighting its potential role in sarcoma progression and the importance of environmental health risk assessment.
Arecoline, the primary alkaloid in areca nut, induces renal injury, yet its underlying molecular mechanisms remain poorly understood. This study aimed to identify arecoline’s molecular targets and elucidate the subsequent pathological signaling cascade in renal cells. We integrated network toxicology, molecular docking, and molecular dynamics (MD) simulations to predict arecoline’s targets. In vitro validation in human kidney (HK-2) cells assessed target engagement via a cellular thermal shift assay (CETSA). Downstream signaling, inflammatory, and fibrotic markers were evaluated using Western blotting, ELISA, and immunofluorescence. Computational models predicted a stable arecoline-Toll-like receptor 4 (TLR4) complex. CETSA demonstrated that arecoline enhanced TLR4 thermal stability in HK-2 cells, strongly indicating target engagement. This binding activated the PI3K/AKT/NF-κB signaling pathway. Consequently, arecoline induced NF-κB-dependent upregulation of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β), elevated the renal injury marker KIM-1, and promoted the accumulation of fibrotic proteins (α-SMA, Collagen III). Arecoline instigates nephrotoxicity by targeting TLR4 and activating the PI3K/AKT/NF-κB axis, driving an inflammatory and fibrotic phenotype in renal tubular cells. The arecoline-TLR4 interaction represents a critical event in areca nut-associated nephropathy, offering a novel therapeutic target.
The introduction of EGFR inhibitors has revolutionized the management of NSCLC, leading to a significant extension of survival in the target patient population. Unfortunately, acquired resistance appears to be unavoidable, which restricts the use of EGFR-TKIs in the clinic. EGFRC797S represents a novel resistant mutation after treatment with the latest generation of EGFR inhibitor Osimertinib. Consequently, developing next-generation EGFR-TKIs targeting such resistant mutations is strongly demanded. Osimertinib-resistant NSCLC cell lines were constructed by transfecting EGFRdel19/T790M/C797S or EGFRL858R/T790M/C797S lentiviral plasmids into NSCLC PC-9 cells, followed by puromycin screening. The drug screening was conducted in a drug repurposing compound library provided by TargetMol. The hit compound (CZC54252) was validated by kinase inhibition, western blot, and molecular docking assays. Its role in overcoming Osimertinib resistance was further evaluated by cellular viability, proliferation, and apoptosis tests. Finally, animal studies were conducted in xenograft models to verify the anti-NSCLC effects of CZC54252 in vivo. The cell models resistant to Osimertinib have been successfully established, and CZC54252 was screened as a hit compound that could overcome Osimertinib resistance, with IC50 values of 0.24 ± 0.06 µM and 0.27 ± 0.09 µM on PC-9del19/T790M/C797S and PC-9L858R/T790M/C797S cells, respectively, which were significantly lower than those of Osimertinib (2.1 ± 0.32 µM and 2.8 ± 0.37 µM, respectively). CZC54252 displayed potent inhibitory activity on EGFRdel19/T790M/C797S (IC50: 1.66 nM) and EGFRL858R/T790M/C797S (IC50: 2.92 nM), and significantly repressed EGFR phosphorylation in western blot assay. Additionally, the pharmacodynamics studies demonstrated that CZC54252 moderately suppressed cell proliferation and induced cell apoptosis both in vitro and in vivo. Notably, not all drug candidates targeting the same molecular pathways as CZC54252 showed activity in the screening, indicating that CZC54252 may overcome Osimertinib resistance by directly targeting EGFRC797S triple mutations rather than its original targets. These data indicate that CZC54252 represents a promising lead compound for overcoming Osimertinib resistance by targeting EGFRC797S, which provides a chemical basis for the development of novel fourth-generation EGFR inhibitors.
The problem of wound healing and the availability of effective and safe agents with regenerative properties is critically important today. The aim of the study was to develop an algorithm for evaluating products with regenerative potential using a set of in vitro and in vivo experimental models. In vitro research included methods of working with cell culture. Studies were conducted using cell lines: HaCat, MAEС, 3T3-A31. In vivo research was conducted on Wistar rats with simulation of the pathological condition of a cut wound and the application of experimental samples and the following assessment of a set of indicators. As a result of using a comprehensive approach to assess the effectiveness of agents with regenerative activity, it was shown that the spray and gel samples, the main component of which is collagen, with decamethoxine, particles of silver sulfadiazine, hyaluronic acid, and additionally a set of amino acids in a spray, were shown to cause a statistically significant increase in the number of live cells at certain concentrations in vitro and increase the speed of restoration of the damaged monolayer of endothelial cells. According to the results of the speed of healing of the skin in the area of the simulated cut wound, a statistically significant increase in the speed of healing from 10 to 19
This study aimed to investigate the wound-healing potential of syringic acid (SA) through integrated in silico and in vitro approaches using human dermal fibroblasts (HDFs). Network pharmacology analysis identified myeloperoxidase (MPO), a key enzyme involved in oxidative stress and inflammation, as a primary target of SA. Computational predictions also indicated favorable pharmacokinetic and ADMET properties. In vitro experiments were conducted using SA at concentrations of 50–400 µM. Cell viability was assessed by MTT and LDH assays, while antioxidant and inflammatory responses were evaluated by measuring SOD, CAT, MDA, TNF-α, IL-1β, TGF-β, iNOS, and hydroxyproline levels. SA treatment significantly enhanced fibroblast viability and exhibited strong antioxidant effects, as demonstrated by increased SOD and CAT activities and reduced MDA levels. Additionally, SA suppressed pro-inflammatory mediators (TNF-α, IL-1β, and iNOS) while upregulating TGF-β expression. Importantly, SA promoted collagen synthesis, evidenced by increased hydroxyproline content. Among the tested doses, 100–200 µM showed the most pronounced effects. Overall, these findings demonstrate that SA facilitates wound healing by improving cellular viability, reducing oxidative stress and inflammation, and enhancing collagen production. This study highlights SA as a promising multi-target therapeutic candidate for wound repair, although further in vivo and safety studies are required to support its clinical application.
Abstract Background 3,4-Methylenedioxymethamphetamine (MDMA) is a psychostimulant known for its social and empathogenic effects. However, its long-term impact on sexual performance and reproductive functions remain controversial. Objective In this study, the effects of chronic MDMA exposure and subsequent withdrawal on sexual behaviour, reproductive hormones, sperm parameters, oxidative stress, systemic inflammation and testicular histology in male rats were investigated. Methods Fifteen male rats were divided into three groups (n = 5 rats/group): control (distilled water-treated), MDMA-treated (100 mg/kg orally for 30 days), and MDMA-withdrawal (100 mg/kg of MDMA for 30 days followed by 30-day drug-free recovery). Sexual behaviour was assessed every ten days for 60 days. Epididymal sperm suspension analysis, serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, markers of oxidative stress and inflammation interleukins 1beta, 6, and 10 (IL-1β, IL-6, IL-10), tumor necrosis factor-alpha (TNF-α) in serum and testes, and histological evaluations of testes were performed. Results MDMA significantly enhanced sexual behavioural parameters (mount, intromission, and ejaculation frequencies) during exposure, but these declined with drug withdrawal. Sperm count and motility decreased following MDMA treatment, with partial recovery after withdrawal. Oxidative stress marker (malondialdehyde, MDA) and proinflammatory cytokines (IL-1β, IL-6, TNF-α) were significantly increased while antioxidant enzymes [superoxide dismutase (SOD), catalase, total thiol] were significantly decreased both in the serum and testes. MDMA also suppressed LH, FSH, and testosterone levels, while histological analysis showed reduced spermatid populations, which improved post-cessation. Conclusion MDMA elicits prosexual effects that was accompanied by low epididymal sperm quality, oxidative stress, inflammation, and hormonal suppression during exposure. Nonetheless, partial recovery occurred following MDMA cessation, suggesting potential long-term reproductive risks.
Myocardial ischemia-reperfusion injury (MIRI) is a major contributor to global disease mortality. This study aims to investigate the protective mechanism of the RMRP/miR-206 axis in MIRI following sevoflurane (Sev) preconditioning. In this study, a hypoxia/reoxygenation (H/R) model was constructed by H9c2 cells. The optimal concentration of Sev was selected by assessing cell viability with a CCK-8 assay. After Sev preconditioning, flow cytometry was employed to detect cell apoptosis. The content of LDH was measured with a lactate dehydrogenase (LDH) assay kit. The SOD activity was examined by an SOD assay kit, and the levels of IL-6 and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA). RT-qPCR was utilized to detect the RMRP and miR-206 expression. The luciferase reporter assay and RIP assay were employed to demonstrate the interaction between RMRP and miR-206. In this study, 2.0
Di(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer and endocrine disruptor that can impair ovarian function by provoking oxidative stress, mitochondrial injury, and apoptosis in granulosa cells. Lycopene is a diet-derived carotenoid with antioxidant and mitochondria-protective activities. This study investigated whether lycopene protects human granulosa-like KGN and COV434 cells against DEHP-induced damage and whether SIRT1 mediates these effects. Cells were exposed to DEHP (50–800 µM) for 24–72 h to define concentration- and time-dependent cytotoxicity, followed by co-treatment with lycopene (1–3 µM). Cell viability was assessed using CCK-8, and apoptosis was evaluated by caspase-3/-7 activity. Intracellular reactive oxygen species (ROS) were quantified using DCFH-DA fluorescence. Mitochondrial function was assessed by JC-1 staining for mitochondrial membrane potential and by qPCR determination of mitochondrial DNA (mtDNA) copy number. Antioxidant defense and lipid peroxidation were evaluated by measuring superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) levels. SIRT1 expression was examined at the transcript and protein levels, and siRNA-mediated SIRT1 knockdown was used to test mechanistic dependence. DEHP significantly reduced cell viability and increased caspase-3/-7 activity in both cell lines, accompanied by ROS accumulation, loss of mitochondrial membrane potential, reduced mtDNA copy number, decreased SOD/CAT activities, and elevated MDA. Lycopene markedly mitigated these effects, restoring redox balance, antioxidant capacity, and mitochondrial integrity while suppressing apoptotic signaling. DEHP downregulated SIRT1, whereas lycopene restored SIRT1 expression, and SIRT1 silencing partially weakened lycopene-mediated cytoprotection. Collectively, these findings identify a SIRT1-dependent protective action of lycopene against DEHP-induced granulosa cell oxidative and mitochondrial injury, supporting lycopene as a potential strategy to mitigate phthalate-associated ovarian toxicity.
Abstract Background Atorvastatin (ATO) is a widely prescribed lipid-lowering drug, but its use can be limited by hepatotoxicity, potentially linked to metabolism-related mechanisms. The cellular pathways connecting ATO metabolism to oxidative imbalance and apoptotic alterations remain incompletely defined. This study examined the roles of oxidative stress, mitochondrial-associated apoptosis, and the pregnane X receptor (PXR)–CYP3A4 axis in ATO-induced hepatic changes, and evaluated whether coenzyme Q10 (CoQ10) could modulate these effects depending on administration timing. Methods Forty male albino rats were randomly assigned to four groups (n = 10). Group I received vehicle (5% DMSO + olive oil), Group II received atorvastatin (ATO, 80 mg/kg), Group III received ATO (80 mg/kg) plus CoQ10 (10 mg/kg), and Group IV received ATO for 21 days followed by CoQ10 for 21 days. All treatments were administered orally once daily. Hepatic outcomes were assessed using biochemical markers, histopathology, and Bcl-2 immunohistochemistry. Microsomal CYP3A1 (rat ortholog of human CYP3A4) catalytic activity was determined using a testosterone 6β-hydroxylation assay. Molecular docking evaluated interactions of ATO, its lactone metabolite, and CoQ10 with CYP3A4 and PXR. Results ATO induced oxidative imbalance, evidenced by increased lipid peroxidation and nitric oxide alongside depleted antioxidant defenses, accompanied by liver structural alterations and reduced Bcl-2 expression. ATO significantly reduced hepatic microsomal CYP3A1 activity to ~ 29% of vehicle control (~ 71% decrease). CoQ10 co-administration markedly attenuated these effects, restoring redox homeostasis and increasing Bcl-2 expression, while co-treatment restored CYP3A1 activity (~ 90% of vehicle control). Post-treatment with CoQ10 partially restored CYP3A1 activity (~ 60% of vehicle control). Docking analysis indicated favorable binding of ATO and its lactone to CYP3A4 and PXR, supporting metabolism-linked bioactivation, while CoQ10 exhibited lower affinity, suggesting indirect modulation. Conclusion ATO-induced hepatic injury is associated with oxidative stress and mitochondrial-related apoptotic processes linked to metabolism-dependent mechanisms. Functional impairment of CYP3A1 activity accompanied ATO-induced oxidative damage, whereas CoQ10 preserved enzymatic activity, consistent with hepatocellular protection. Early CoQ10 intervention provided superior protection, highlighting the importance of timing in mitigating metabolism-related hepatic damage. These findings provide mechanistic insight into statin-induced hepatotoxicity and support further investigation of CoQ10 as a complementary strategy to enhance statin safety.
Breast cancer is the most common cancer affecting women, and plasma-activated water (PAW) is emerging as a potential therapeutic approach. While early results are encouraging, additional research is necessary to assess its long-term effects and identify the most effective methods of application. Therefore, this study aimed to investigate the effects of PAW on an MCF7 breast cancer mouse model. A total of eighteen BALB/c mice were divided into three groups (n = 6/group): one negative control group and two tumor groups injected with MCF7 cells. After 91 days of MCF7 injection, the tumor groups received either distilled water (DW) or PAW (for 3 min) twice weekly via oral gavage for 28 days. The results revealed that the conductivity, H2O2, and NO2- values were significantly higher for PAW than for DW, while pH was lower (P < 0.001). Hematological analysis showed an increase in neutrophil counts, neutrophil
Tumor necrosis factor-alpha (TNF-α) plays a central role in chronic inflammatory diseases. Anti-TNF agents are widely used in rheumatological conditions; however, their association with thyroid hormone parameters in patients without pre-existing thyroid disease remains incompletely understood. This study aimed to evaluate changes in thyroid hormone profiles during anti-TNF therapy in euthyroid patients with rheumatic diseases. In this retrospective study, 98 patients diagnosed with rheumatoid arthritis, ankylosing spondylitis, or Behçet’s disease without known thyroid disease were evaluated. Thyroid function tests, including thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), and free thyroxine (fT4), anti-thyroid peroxidase (anti-TPO) antibodies, inflammatory markers such as C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR), and metabolic parameters were assessed at baseline and after 3 and 6 months of anti-TNF therapy. Anti-TNF therapy was associated with significant reductions in inflammatory markers (CRP and ESR, p < 0.01). A modest decrease in fasting glucose levels and an increase in high-density lipoprotein cholesterol (HDL-C) were observed during follow-up (p = 0.024 and p = 0.044, respectively). TSH and fT4 levels remained stable over time, whereas a gradual increase in fT3 levels was observed (p < 0.01). No significant changes were detected in anti-TPO antibody levels. Among euthyroid patients with rheumatic diseases, predominantly rheumatoid arthritis and ankylosing spondylitis, anti-TNF therapy was associated with stable thyroid function parameters. The observed increase in fT3 levels may reflect reduced inflammatory burden rather than direct thyroidal effects. These findings support the thyroid safety of anti-TNF agents while highlighting potential links between inflammation control and peripheral thyroid hormone conversion.
Metastasis and chemoresistance remain key challenges in colon tumor management. Integrating transcriptomic screening with molecular modeling may reveal actionable therapeutic targets for rational nanocomplex design. Transcriptomic datasets GSE44076 (n = 98) and GSE253699 (n = 6) were analyzed using limma (adjusted p < 0.05, |log₂FC| > 1). Differentially expressed genes were intersected with curated metastasis and chemoresistance panels which resulted in 18 overlapping targets. Hub genes were identified via STRING and Cytoscape-BiNGO, and prognostic analysis using KMplotter highlighted PDCD4 and CCND1 as favorable markers. A PTX–TP@PLGA–iRGD nanocomplex was designed by co-encapsulating paclitaxel and Trametes robiniophila Murr–derived metabolites within a PLGA matrix functionalized with iRGD. Nanocomplexes were characterized by dynamic light scattering, zeta potential, and HPLC for size, stability, and drug encapsulation. Functional validation included docking, 50 ns MD simulations, in vitro cytotoxicity uptake, and target engagement assays. PDCD4 and CCND1 emerged as convergence nodes, with PDCD4 showing stage-wise downregulation (p = 0.014) and positive correlation with immune infiltration. The nanocomplex exhibited a uniform particle size ( 165 nm, PDI 0.12), and encapsulation efficiency > 75
Abstract Background KRAS mutations are approximately 25% of human cancers, with particularly high incidence in breast and lung cancers, by constitutively triggering MAPK/ERK and PI3K/AKT pathways that advance proliferation, survival, and autophagy-mediated resistance. Targeting these pathways with kinase inhibitors upregulates autophagy, thereby diminishing therapeutic efficacy. Thus, combining kinase inhibitors with autophagy inhibitors offers a rational strategy to enhance antitumor benefits in KRAS-mutant malignancies. FDA-approved Sorafenib (multi-kinase inhibitor targeting Raf) and Hydroxychloroquine (autophagy inhibitor) show promise for repurposing, as Sorafenib induces autophagy leading to resistance, warranting combination testing in KRAS-mutant models. This study integrates computational modeling and in vitro assays to evaluate their synergistic potential in MDA-MB-231 breast and A549 lung cancer cells. Methods Raf-Sorafenib stability was evaluated using RMSD, RMSF, Rg, hydrogen bonds, contact frequency, and MM/GBSA for binding free energy in molecular dynamics simulations (100 ns, GROMACS with CHARMM36 force field). MTT assays were used for cytotoxicity on MDA-MB-231, A549, and normal gingival fibroblasts (48 h treatment); Chou-Talalay Combination Index and Dose Reduction Index were used for synergy; Annexin V/PI flow cytometry was performed for apoptosis; PI staining was used for cell cycle; and ANOVA/Tukey’s test (GraphPad Prism, p < 0.05) was conducted for statistics. Results Sorafenib bound Raf stably (RMSD ~ 0.25 nm protein/0.15 nm ligand, G_bind − 49.90 ± 2.69 kcal/mol), with persistent interactions (3–4 H-bonds, key residues VAL471, LEU513). IC50 values: Sorafenib 9.4 µM (MDA-MB-231), 12 µM (A549), 23.1 µM (fibroblasts); HQ 23.6/62.4/86.2 µM; SN:2HQ ratio showed synergy in MDA-MB-231 (CI = 0.32, DRI 9.36 Sorafenib/4.68 HQ at Fa = 0.5) but antagonism in A549 (CI > 1). Combination enhanced late apoptosis/necrosis (49.41%) in MDA-MB-231 with minimal normal cell cycle disruption. Conclusions Sorafenib-HQ combination offers potent, context-specific synergy for KRAS-mutant breast cancer via Raf inhibition and autophagy blockade, enabling dose reductions and apoptosis enhancement. Tumor-type dependence (synergy vs. antagonism) highlights need for patient stratification; findings support repurposing with limited normal cell impact.
Abstract Background Inflammation serves as a natural defense mechanism; however, its persistence can lead to chronic diseases with serious clinical consequences. Early detection of inflammation is therefore critical to slowing disease progression and improving therapeutic outcomes. Methods In this study, cefaclor (Cefa) was successfully radiolabeled with iodine-131 via electrophilic substitution to facilitate the detection of infected and inflamed muscles in mouse models. The labeling reaction was carried out with 100 µg of Cefa and 100 µg of iodogen, using glass frits as the oxidizing system at pH 7 and 60 °C, with 10 µL of Na131I for 20 min. The resulting [131I]Cefa was purified by high-performance liquid chromatography (HPLC). Molecular modeling was performed in the Molecular Operating Environment (MOE) to evaluate the compound’s structure and binding affinity. Results The labeling process was optimized to achieve a radiolabeling efficiency of 90 ± 0.56%, and stability of about 89 ± 0.5% at 4 h. Docking simulations confirmed strong binding of [131I]Cefa to bacterial DNA gyrase B, supporting its potential as a targeted imaging agent. Biological evaluation in mouse models demonstrated notable tracer accumulation in both septic and sterile inflammatory sites. Uptake values reached 28 ± 1.5%ID/organ in infected muscles and 16 ± 1.5%ID/organ in sterile inflammation at 120 minutes post-injection. The target-to-non-target (T/NT) ratios were 5.28 for infected muscles and 2.1 for sterile inflammation, indicating effective differentiation between bacterial (septic) and non-bacterial (aseptic) inflammatory foci. Conclusion The radiolabeled [131I]Cefa compound exhibits promising diagnostic capabilities for distinguishing bacterial infections from sterile inflammation. Its high radiolabeling efficiency, strong molecular binding, and selective in vivo uptake support its potential utility as a non-invasive imaging agent for early detection and characterization of inflammatory conditions.
Antipyretic drugs are widely used to manage fever and pain and are generally regarded as safe when administered within recommended therapeutic ranges. Nevertheless, concerns remain regarding the potential biological effects associated with repeated or sub-chronic exposure. The present study evaluated the hematological, histopathological, and molecular effects of two commonly used antipyretics, paracetamol and ibuprofen, following repeated administration in male albino mice. Hematological analysis revealed drug-related alterations in selected blood parameters, with more pronounced changes observed in ibuprofen-treated groups, while paracetamol exposure was associated with comparatively milder effects. Qualitative histopathological examination demonstrated organ-specific structural alterations in the testes, liver, kidneys, and stomach, which were generally mild in nature and more frequently observed at higher experimental dose levels. No evidence of overt testicular failure or severe tissue damage was detected within the exposure period. At the molecular level, repeated exposure to paracetamol and ibuprofen was associated with downregulation of TNF-α and connexin 43 mRNA expression in peripheral blood, suggesting early systemic transcriptional responses accompanying the observed histological changes. However, molecular findings were limited to mRNA expression analysis and did not include protein-level or tissue-specific validation. Overall, the results indicate that repeated sub-chronic exposure to paracetamol and ibuprofen at therapeutic-equivalent dose ranges may be associated with early hematological, histological, and molecular alterations in mice. These findings highlight the importance of cautious and rational use of widely prescribed antipyretic drugs and underscore the need for further studies to elucidate long-term safety, underlying mechanisms, and tissue-specific functional outcomes.
Neurodegenerative disorders and behavioural abnormalities, including learning and memory issues, are significantly influenced by infection and inflammation. We looked into how 7-hydroxyflavone affected a mouse model of learning and memory impairment brought on by LPS. This investigational protocol aims to assess the impacts of 7HF on LPS-induced neuroinflammation and its impact on memory impairment in mice. Mice were pretreated with 7HF (5, 10 mg/kg, po) and received LPS (250 µg/kg ip) for 7 days. The Morris water maze (MWM) and the NORT were used for assessing memory. In order to analyse the oxidative stress markers, biochemical assessments were performed after the mice were euthanised. Moreover, neuroinflammatory markers in the hippocampi of mouse brains, such as IL-6 and NFKB, were estimated. LPS administration led to decreased memory retention in both MWM and NORT, as a distinct rise in the oxidative stress, including lipid peroxidation (LPO) and reduction in glutathione (GSH) levels, was also observed. Moreover, reductions in IL-6 and NFKB levels were observed. Pretreatment of animals with 7HF reversed the LPS-induced behavioural and memory impairments, and it was also found to lower the IL-6 and NFKB levels in the hippocampus. This study demonstrates how 7HF has neuroprotective properties that help prevent memory loss and neuroinflammation brought on by LPS.
Abstract Objective To study the potential cardioprotective role of dapagliflozin in comparison to metformin in experimentally- induced myocardial infarction in rats with type 2 diabetes mellitus (T2DM). Materials and Methods This study was comparative study, which was housed at the animal house, Mansoura Experimental Research Center (MERC). Fifty adult male albino rats were included in this study and there were further randomly divided into Group-I: (Normal), Group –II: rats received Streptozotocin (STZ) for induction of type 2 DM. This group was subdivided into (4) equal subgroups as the following: Group –IIa: (DM), Group-IIb: (DM+ISO), Group–IIc: (DM+ISO+Metformin), Group-IId: (DM+ISO+DAPA). The outcomes of the study were measured by ECG recording, serological study which included Interleukin (IL)-10, IL6, and tumor necrosis factor (TNF)-α, troponin, lipid profile and blood glucose and histopathological examination of isolated hearts. Results Dapagliflozin show better cardioprotective effect than Metformin with lower IL-10, IL-6, TNF, CTNI and Troponin C levels than Metformin with statistically significant difference (p < 0.001*). The percentage of area of edema and degeneration is the lowest in controls followed by DM+ISO+DAPA group then DM+ISO+Metformin group when compared with DM and DM+ISO groups which revealed the cardio protective effect of DAPA and Metformin with inter-groups statistically significant difference (p < 0.001*) Conclusion Dapagliflozin has cardioprotective effects against myocardial infarction in type 2 diabetic rats. This effect might be due to its antifibrotic and antiarrhythmic effects. However, Metformin did not show any antifibrotic or anti-arrhythmic effects.
