The liver is a decisive organ that is damaged by sepsis. Medicarpin (Med) is a principal wing-type final product of 5-deoxyisoflavone sub-branch in M. truncatula, and exerts anti-tumor and anti-inflammatory effects in diverse diseases. This research aimed to elucidate the Med function and the prospective mechanism in sepsis-induced liver injury. After treating THLE-2 cells with diverse Med concentrations (0, 5, 10, 20, 40, 80, 160 μM), Med toxic effect on THLE-2 cells was checked through a Cell Counting Kit-8 (CCK-8) experiment. Based on a sepsis-induced liver injury model using lipopolysaccharide (LPS) and THLE-2 cells, Med function in sepsis-induced liver injury was verified via CCK-8 assay, Western blot, immunofluorescence, and ELISA. Also, Med mechanism in sepsis-induced liver injury was examined via Western blot, CCK-8 assay, and ELISA. Furthermore, Med role in sepsis-induced liver injury in vivo model was elucidated through animal experiments, hematoxylin-eosin staining, ELISA, Western blot, and immunofluorescence. Med possessed no obvious impact on THLE-2 cell viability when Med concentration was lower than 160 μM. Med alleviated repression of LPS-induced THLE-2 cell viability, and also reduced THLE-2 cell pyroptosis induced by LPS. From a mechanistic perspective, our data further illustrated that LPS induced a decrease in THLE-2 cell viability, and Med enhanced cell viability, while this impact was partially reversed after NLRP3 overexpression, indicating that Med partially counteracted the effects of NLRP3 overexpression on cell viability. Meanwhile, in vivo experimental data further demonstrated that Med alleviated liver injury and pyroptosis in sepsis mice, mainly through reduction of ALT and AST levels, a decrease in liver injury score of mice, a reduction in IL-1β and IL-18 levels, as well as a decrease in NLRP3, ASC, cleaved caspase-1, and GSDMD-N protein levels. In conclusion, Med exerted its protective effect on sepsis-induced liver injury through NLRP3/GSDMD/caspase-1-mediated cell pyroptosis.
Hepatotoxicity is one of the most crucial side effects of chemotherapy administration. Obeticholic acid (OCA) is a semisynthetic bile acid and farnesoid X receptor (FXR) agonist derived from chenodeoxycholic acid, with reported antioxidant and anti-inflammatory effects in liver disorders. This study investigated the hepatoprotective effect of OCA against commonly used chemotherapy cisplatin (CP)-induced hepatotoxicity in rats, as well as the modulatory effects of edaravone (EDA), a potent free radical scavenger, on its effects. Rats were divided into five groups: control (received vehicle), CP (7.5 mg/kg), EDA (30 mg/kg) + CP, OCA (30 mg/kg) + CP, and EDA + OCA + CP. The results of the present study demonstrated that both OCA and EDA significantly mitigated liver damage caused by CP, as evidenced by restoring liver enzymes and histological structure, reestablishment of oxidant/antioxidant status, suppression of inflammation, and attenuation of pro-death signaling. The study highlights the role of key molecular pathways, including Keap1/Nrf2/HO-1,HO-1, TNF-α/NF-κB, and AKT/GSK-3β, in the hepatoprotective mechanisms of OCA. Collectively, these findings suggest that OCA and EDA, particularly in combination, attenuate CP-induced hepatotoxicity and are associated with coordinated modulation of oxidative stress, inflammatory signaling, and AKT/GSK-3β-associated pro-survival/pro-death pathways.
Elucidating the molecular distinctions between invasive and non-invasive forms of lung adenocarcinoma is essential for the development of personalized therapeutic approaches and the optimization of patient outcomes. Transfer RNA-derived small RNAs (tsRNAs) have been recognized as pivotal regulators in oncogenic processes. In this study, we identified a specific tsRNA associated with non-invasive lung adenocarcinoma and demonstrated its suppressive effects on cellular proliferation and metastatic potential. At the molecular level, this tsRNA exerts its regulatory function by targeting and repressing G6PD mRNA, thereby perturbing the equilibrium between NADPH and reactive oxygen species (ROS) production. This discovery positions the identified tsRNA as a promising candidate for therapeutic intervention in non-invasive lung adenocarcinoma. Further investigation into the role of tsRNAs in lung adenocarcinoma is imperative to fully harness their potential in clinical management.
The widespread accumulation of polystyrene nanoplastic particles (NPPs) in the environment has raised significant human health concerns, specifically on immune function. However, the underlying molecular mechanisms behind NPP induced immune cell (monocyte/macrophage) toxicity and their relationship with existing pathological conditions remain largely unexplored. The current study explored the interaction of NPPs with serum proteins and composition of protein corona through proteomic analysis (LC-HRMS/MS). Further, we investigated the immunotoxic effects on THP-1 macrophage cells via various cellular and molecular assays, including cytotoxicity, oxidative stress, mitochondrial function, intracellular calcium homeostasis, apoptosis and stress signaling pathway assessment. This study further assessed the amplification potential of NPPs on inflammatory (TNF α and lipopolysaccharide), atherogenic (ox-LDL) and environmental (polycyclic aromatic hydrocarbons (PAH)) stress conditions. The findings demonstrated that NPPs triggered cytotoxicity in a dose dependent manner along with excessive ROS generation, mitochondrial impairment, elevated intracellular calcium levels and apoptosis by altering BAX and BCL2 expression. We also found that NPPs activated stress signaling pathways through modulating HSP27, SAPK/JNK, p38 MAPK and c-Jun phosphorylation. Interestingly, pathway inhibition studies confirmed the involvement of p38 MAPK, ERK and mitochondrial ROS signaling in NPP-associated toxicity. In addition, NPPs exposure augmented inflammatory response, promoted atherogenesis (foam cell formation), amplified PAH induced toxicity. Together, these observations revealed precise adverse outcome pathways (AoP) and stress amplification effects of NPPs on macrophages and consequent toxic effects on the immune system.
Mesenchymal stem cells (MSCs) hold considerable therapeutic promise in regenerative medicine, yet precise control over lineage fate remains critical for reproducible clinical outcomes. This review examines the Nrf2-Keap1 (nuclear factor erythroid 2-related factor 2-Kelch-like ECH-associated protein 1) signaling axis as a redox-sensitive rheostat governing the balance between osteogenic and adipogenic differentiation. Physiological reactive oxygen species (ROS) support osteogenesis in a dose- and context-dependent manner, whereas sustained oxidative stress drives cellular senescence and adipogenic bias; emerging evidence also indicates that Nrf2 hyperactivation can itself impair osteogenic differentiation, pointing to an optimal activation window rather than a simple "more is better" relationship. Conventional plant-derived Nrf2 activators are limited by poor bioavailability and metabolic stability. Phytofabricated metal oxide nanoparticles (MONPs), synthesized using plant extracts as capping and stabilizing agents, offer a biocompatible, "green" delivery alternative. We propose that these hybrid systems may act through a dual-stimulus mechanism - phytochemical-mediated electrophilic modification of Keap1 cysteines alongside metal-oxide-core-derived, sub-cytotoxic ROS consistent with mitohormetic signaling - that together could promote Nrf2 nuclear translocation, ARE-driven antioxidant gene expression, and downstream epigenetic reinforcement of osteogenic commitment. This dual-stimulus framework, however, remains a mechanistic hypothesis: no study to date has directly demonstrated Keap1 modification, Nrf2 activation, and resulting lineage bias by phytofabricated MONPs within MSCs. Preclinical studies support the efficacy of phytofabricated MONPs in bone defect models and suggest potential for restoring function in senescent MSCs, but direct mechanistic validation in MSC systems remains an essential next step before these insights can guide rational nanoplatform design for bone regeneration.
Uterine leiomyomas (UL) represent one of the most prevalent gynecological benign tumors with a considerable global medical and economic burden. Effective therapeutic strategies that preserve the uterus and maintain fertility are still lacking. Fisetin (FIS), a naturally occurring dietary flavonoid, exhibits antitumor, anti-inflammatory, and antifibrotic properties, yet its therapeutic potential in UL remains incompletely defined. This study investigated the therapeutic effects of fisetin on UL and the possible underlying mechanisms. UL was experimentally induced in adult female rats by monosodium glutamate (MSG), and animals were assigned to Control, FIS, UL, and UL + FIS groups. Blood and tissue samples were subjected to hormonal, biochemical, histopathological, and immunohistochemical assessments. Serum estradiol, progesterone, and S1P levels were measured using ELISA, while qPCR quantified the expression of key signaling molecules, including SphK1, TGF-β1, HMGB1, TNF-α, and IL-1β. Immunohistochemistry assessed TGF-β1 and α-SMA expressions, and histopathology (H&E and Masson's Trichrome stains) evaluated tissue architecture and fibrosis. The results showed that fisetin administration significantly reduced the elevated sex hormones, inhibited S1P signaling, and suppressed proinflammatory mediators (HMGB1, TNF-α, IL-1β) and profibrotic mediators (TGF-β1, α-SMA). Histopathological evaluation confirmed decreased fibrotic remodeling and improved tissue architecture. Overall, these results suggest that fisetin's effect may involve coordinated modulation of the S1P/SphK1, TGF-β, and HMGB1 axes, highlighting its potential as a novel uterus-preserving therapeutic agent for UL.
Duloxetine, a serotonin-norepinephrine reuptake inhibitor, commonly prescribed for depression and chronic pain, has been implicated in hepatotoxicity, primarily driven by oxidative stress and inflammatory responses. This has led to growing interest in naturally derived antioxidant compounds as potential protective strategies. The aim of this study was to evaluate the hepatoprotective potential of nanocurcumin against duloxetine-induced liver injury in a controlled animal model. Male Wistar rats were divided into eight experimental groups, control, Solvent, curcumin 200 mg/kg, nanocurcumin 200 mg/kg, duloxetine 60 mg/kg, and treatment groups that simultaneously received duloxetine and nanocurcumin. Biochemical assays measured liver function enzymes (ALT, AST), oxidative stress markers (MDA, TOS), and antioxidant enzymes (SOD, GPx). Proinflammatory cytokine levels, TNF-α were also measured and liver tissue was evaluated histologically. Duloxetine administration significantly increased liver enzymes, oxidative stress markers, and TNF-α levels, along with histopathological evidence of liver injury. Concomitant treatment with nanocurcumin reduced these effects and attenuated these changes, as evidenced by normalization of biochemical indices, enhanced antioxidant defense, suppression of proinflammatory cytokines, and preservation of liver structure. Nanocurcumin exhibits potent hepatoprotective effects against duloxetine-induced liver injury, likely through enhancement of antioxidant capacity and modulation of inflammatory responses. These findings support its potential as an adjunctive therapy to duloxetine to mitigate hepatic side effects.
BDNF and its receptor, TrkB, are essential regulators of energy homeostasis in both central and peripheral tissues, including the liver. This study aimed to investigate the effects of ANA-12, a selective TrkB antagonist, on hepatic mitochondrial enzyme activities and biochemical parameters in a mouse model of sucrose-induced insulin resistance. Male C57BL/6J mice were divided into four groups: Control, Sucrose (35% solution), Sucrose+DMSO, and Sucrose+ANA-12 (0.5 mg/kg). Following a 16-week dietary intervention, ANA-12 was administered via intraperitoneal injection during the final 21 days. Hepatic activities of cytochrome c oxidase, superoxide dismutase (SOD), and catalase (CAT), along with malondialdehyde (MDA) levels, were analyzed to evaluate mitochondrial enzyme activity and oxidative stress-related biochemical changes. Chronic sucrose consumption significantly increased body weight, serum insulin levels, HOMA-IR scores, and cholesterol levels. While ANA-12 treatment showed a numerical tendency to attenuate weight gain and HOMA-IR scores, these changes did not reach statistical significance. However, the ANA-12 group exhibited significantly lower serum leptin and ceramide levels compared to the Sucrose+DMSO group. In the liver, sucrose consumption triggered oxidative stress, as evidenced by increased MDA levels. Conversely, TrkB inhibition was associated with increased cytochrome c oxidase activity, altered SOD activity, and reduced hepatic MDA concentrations compared with the Sucrose+DMSO group, suggesting partial modulation of oxidative stress-related biochemical alterations. Furthermore, chronic sucrose consumption was associated with reduced hepatic BDNF levels. Overall, these findings indicate that TrkB inhibition may influence selected hepatic mitochondrial enzyme activities and redox-related markers under sucrose-induced metabolic stress. However, these results should be interpreted cautiously because systemic insulin resistance markers were not significantly improved, and additional control groups are required to clarify ANA-12-specific and vehicle-related effects.
Cisplatin is a widely used chemotherapeutic agent; however, its clinical utility is frequently limited by dose-dependent adverse effects, particularly hepatotoxicity and nephrotoxicity. Luteolin, a naturally occurring flavonoid abundant in various medicinal plants, exhibits well-documented antioxidant and anticancer activities. This study investigated whether Luteolin isolated from Pistacia terebinthus fruits could attenuate Cisplatin-induced organ toxicity while enhancing its antiproliferative effects. For the in vivo experiments, 48 male Swiss albino mice (9-10 weeks old, 26-30 g) were randomly assigned to eight groups (n = 6/group): control, Cisplatin, Luteolin (25, 50, or 100 mg/kg), and Cisplatin plus Luteolin (25, 50, or 100 mg/kg). Cisplatin (10 mg/kg/day, intraperitoneally) and/or Luteolin (25-100 mg/kg/day, orally) were administered for 14 consecutive days. Serum biochemical parameters, oxidative stress markers, antioxidant enzyme activities, and tissue injury-related biomarkers were evaluated in liver and kidney tissues. Cisplatin administration significantly increased serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, and glucose levels, whereas co-treatment with Luteolin markedly attenuated these alterations in a dose-dependent manner. In addition, Luteolin restored Cisplatin-induced disturbances in oxidative stress and antioxidant defense markers, including malondialdehyde (MDA), 8-hydroxy-2'-deoxyguanosine (8-OHdG), HSP70 levels, superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST) activities in liver and kidney tissues. To support the in vivo findings, in vitro antiproliferative assays were performed using Saos-2 osteosarcoma, HTB-9 bladder cancer, A549 lung cancer, and BEAS-2B normal epithelial cell lines. Cells were treated with Luteolin (5-200 µM) alone or in combination with Cisplatin IC50 concentrations for 24-48 h, and cell viability was assessed using the WST-1 assay. Co-treatment with Luteolin significantly enhanced Cisplatin-induced growth inhibition in cancer cells while exerting limited cytotoxicity in normal cells. Luteolin alone also demonstrated concentration-dependent antiproliferative activity. In conclusion, Luteolin exerted marked hepatorenal protective effects and enhanced the antiproliferative activity of Cisplatin in experimental models. These findings suggest that Luteolin may represent a promising adjunctive candidate for reducing Cisplatin-associated toxicity while improving therapeutic responsiveness.
Acne inversa (AI) or Hidradenitis suppurativa is a chronic, inflammatory disorder characterized by painful nodules, abscesses, and scarring, primarily affecting apocrine gland-rich areas. Despite advances in understanding its clinical features, the molecular mechanisms driving AI remain incompletely defined. Emerging evidence highlights the critical role of microRNAs (miRNAs), small non-coding RNAs that regulate gene expression post-transcriptionally, in modulating inflammatory and immune pathways implicated in AI pathogenesis. Specific miRNAs are dysregulated in AI lesional skin and systemic circulation, influencing cytokine production, immune cell recruitment, and keratinocyte activation. These findings position miRNAs as promising non-invasive biomarkers for early diagnosis and disease monitoring. Furthermore, targeting miRNA expression offers a novel therapeutic strategy to restore immune homeostasis and prevent disease progression. This review synthesizes current knowledge on miRNA involvement in AI, underscoring their potential to revolutionize personalized diagnosis and treatment approaches for this complex inflammatory disease.
Acute pancreatitis is an inflammatory disease of the pancreas that is often accompanied by various complications, which seriously impair patient health. Fibulin-1 (FBLN1) is an extracellular matrix (ECM) protein that plays a crucial role in several cellular processes. Previous quantitative analyses of plasma protein profiles of severe acute pancreatitis patients and healthy volunteers have shown that the expression of FBLN1 is decreased in severe acute pancreatitis patients. However, the regulatory roles of FBLN1 in patients with acute pancreatitis have not been fully investigated. In this study, FBLN1 was found to be expressed at lower levels in acute pancreatitis. Moreover, FBLN1 inhibited Caerulein-triggered inflammatory injury in AR42J cells. It was also found that FBLN1 suppressed Caerulein-triggered ferroptosis in AR42J cells, and that FBLN1 could activate the NRF2/HO-1 pathway. In conclusion, FBLN1 may relieve Caerulein-induced acute pancreatitis in AR42J cells by inhibiting ferroptosis, providing insights on the use of FBLN1 in acute pancreatitis treatment.
Myocardial infarction (MI) usually results in severe heart failure (HF), and patients suffer from the repercussions of MI, elucidation of the pathological basis of MI is crucial to optimize the prevention and treatment of MI. This study investigated the role of dysregulated circ_0001112 in MI. In our study, circ_0001112 was significantly upregulated in the MI model and by hypoxia treatment. Circ_0001112 overexpression inhibited hypoxia-induced cardiomyocyte apoptosis. Luciferase assay showed that circ_0001112 sponged miR-324-5p that regulated the expression of cyclin-dependent kinase 6 (CDK6). Ago2-miRNP immunoprecipitation further confirmed that the 3'-untranslated region (3'UTR) of CDK6 was targeted by miR-324-5p. Either miR-324-5p overexpression or CDK6 knockdown prevented cardiomyocyte apoptosis induced by circ_0001112 overexpression. In conclusion, this study demonstrated that the downregulation of circ_0001112 promoted cardiomyocyte apoptosis by sponging miR-324-5p and upregulating CDK6 expression in MI.
Bone marrow mesenchymal stem cells (BMSCs) prominent capacity for tissue repair and regenerative applications due to their multipotent differentiation potential and immunomodulatory activities. This study explores the function of BMSCs in tendon repair, with a particular focus on their regulatory effect on fat mass and obesity-associated protein (FTO)-mediated m6A methylation of Elongin B (ELOB). In this study, BMSCs were isolated from mice with experimental tendon injury and characterized by detecting the expression of surface biomarkers (CD90, CD44, CD29, CD34, CD45). The proliferation, migration, apoptosis, and intracellular reactive oxygen species (ROS) levels of tenocytes were assessed using CCK-8 assay, wound healing assay, and flow cytometry. RT-qPCR and Western blot analysis were applied to quantify gene and protein expression, respectively. Dual luciferase reporter assay and methylated RNA immunoprecipitation (MeRIP) were performed to verify the regulatory relationship between FTO and ELOB. Functional recovery of injured tendons in mice was evaluated via behavioral tests and hematoxylin and eosin (HE) histological staining. The results showed that BMSCs effectively facilitated tendon repair in the murine tendon injury model. The RNA demethylase FTO was markedly downregulated in injured tendon tissues, and its expression could be modulated by BMSCs. Mechanistically, BMSCs upregulated FTO to accelerate tenocyte proliferation and migration, and simultaneously suppress oxidative stress during tendon injury. FTO repressed ELOB expression through m6A modification. Functional assays further demonstrated that FTO improved tenocyte proliferation and migration, and alleviated oxidative stress by downregulating ELOB. In vivo assays validated that overexpression of FTO boosted tendon repair by reducing ELOB expression. In conclusion, BMSCs improve tendon repair by stimulating tenocyte proliferation and migration and alleviating oxidative stress. This protective mechanism is dependent on the regulation of FTO-mediated m6A methylation of ELOB.
2-Hydroxyethyl methacrylate (HEMA), a monomer commonly used in dental resin materials, has been reported to leach from polymerized resins. However, its potential impact on periodontal tissues remains insufficiently understood. This study investigates the effects of HEMA on human gingival fibroblasts (HGFs), which constitute a major component of periodontal connective tissue. Specifically, we examined whether HEMA induces the production of pro-inflammatory cytokines interleukin-6 (IL-6) and interleukin-8 (IL-8), as well as matrix metalloproteinases (MMP)-1 and MMP-3, which are involved in collagen degradation. Furthermore, we analyzed the intracellular signaling pathways activated by HEMA exposure to elucidate the molecular mechanisms underlying these responses. Cytokine and MMP production were analyzed using the ELISA. In addition, activation of intracellular signaling pathways were examined by Western blot analysis. HEMA treatment did not induce IL-6 or IL-8 production, but significantly increased MMP-1 and MMP-3 levels in a concentration-dependent manner. In contrast, tissue inhibitor of metalloproteinases (TIMP)-1 production was suppressed by HEMA. Moreover, compared with 24-h exposure, 48-h exposure induced increases in MMP production and decreases in TIMP-1 production at lower concentrations of HEMA. Furthermore, HEMA enhanced the phosphorylation of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK), and protein kinase B (Akt). Inhibitor studies revealed that the p38 MAPK, extracellular signal-regulated kinase (ERK), JNK, and Akt signaling pathways are involved in the regulation of MMP-1 and MMP-3 production. These findings suggest that HEMA leaching from resin materials may contribute to periodontal tissue destruction by promoting MMP production in HGFs through activation of specific signaling pathways. Given the potential impact of HEMA on periodontal tissues, it is important to ensure complete polymerization of light-curable resin materials and thorough removal of excess resin cement. The use of HEMA-free materials may also be worth considering to minimize biological risks.
This study explored the role of miR-138-5p in calcific aortic valve disease (CAVD), focusing on its regulation of osteogenic differentiation in human aortic valve interstitial cells (hAVICs) and underlying mechanisms. Aortic valve tissues from CAVD patients and controls were analyzed for miR-138-5p expression using qRT-PCR. hAVICs were cultured in osteogenic medium (OM) and transfected with miR-138-5p mimics or inhibitors. Calcification was evaluated via ALP and Alizarin Red staining. Western blot assessed osteogenic markers (RUNX2, ALP, OPN, BMP-2). Dual-luciferase assays confirmed miR-138-5p targeting of SLC39A14. The role of SLC39A14 and the Nrf2 pathway was examined through overexpression experiments and Western blot for Nrf2, HO-1, and NQO1. ML385 was used to inhibit Nrf2. miR-138-5p was significantly downregulated in CAVD tissues and OM-induced hAVICs. Overexpression of miR-138-5p inhibited calcification, reducing ALP activity, nodule formation, and osteogenic marker expression. Inhibition of miR-138-5p promoted calcification. SLC39A14, upregulated in CAVD, was validated as a direct target of miR-138-5p. Its overexpression promoted calcification, which was reversed by co-transfection with miR-138-5p. Mechanistically, miR-138-5p was associated with activation of the Nrf2 pathway, as evidenced by increased Nrf2 nuclear translocation and HO-1/NQO1 expression, whereas SLC39A14 overexpression showed the opposite effect. Nrf2 inhibition with ML385 diminished the anti-calcific effect of miR-138-5p. miR-138-5p inhibits hAVIC calcification, at least in part through regulation of SLC39A14 and the Nrf2 pathway, highlighting a potentially important regulatory mechanism in CAVD and a candidate therapeutic target for valve calcification.
Acute myocardial infarction (AMI) is closely associated with excessive oxidative stress, inflammation, and activation of the renin-angiotensin system (RAS). The active form of vitamin D [1,25(OH)2D3] and the Nrf2 activator dimethyl fumarate (DMF) exhibit antioxidant and anti-inflammatory effects. This study investigated the individual and combined protective roles of 1,25(OH)2D3 and DMF in an isoproterenol (ISO)-induced model of myocardial injury. Male Sprague-Dawley rats received ISO (85 mg/kg, two doses 24 h apart) and were pretreated with 1,25(OH)2D3 or DMF alone, a combination pretreatment, or a combination post-treatment. Myocardial injury was assessed using serum biomarkers, histopathology, oxidative stress markers (ROS, MDA, GSH), inflammatory markers (TNF-α), RAS components (Ang II, AT1R, ACE), and mRNA expression of Nrf2, NQO1, and NF-κB. ISO administration caused marked myocardial injury, oxidative imbalance, and increased inflammatory and RAS activation while suppressing Nrf2 and NQO1. Both 1,25(OH)2D3 and DMF significantly reduced oxidative stress, inflammation, and components of the RAS. Combination pretreatment and post-treatment further improved redox status and molecular markers. These groups showed elevated Nrf2 and NQO1 expression and reduced NF-κB, AT1R, and ACE expression compared with ISO controls. However, synergistic enhancement was not observed. 1,25(OH)2D3 and DMF provide both preventive and therapeutic protection against ISO-induced myocardial injury by modulating oxidative stress, inflammation, the Nrf2 and NF-κB signaling pathways, and RAS regulation.
Cyclophosphamide (CYP) is implicated in oxidative stress (OS)-related platelet destruction during chemotherapy, leading to thrombocytopenia. Antioxidants can potentially attenuate the side effects of drugs and improve therapeutic outcomes. Melatonin is highly permeable and has radical scavenging and antioxidant enzyme-stimulating activity. This study investigates the influence of melatonin on platelets during CYP-induced OS under ex vivo conditions. Platelets isolated from the whole blood of male Wistar rats (n = 5) were categorized and treated as follows: Controls (CON) and Experimentals: (i) CYP, (ii) melatonin (MEL), and (iii) CYP+melatonin (CYP + MEL). Antioxidant defenses, OS and platelet function markers were assessed. Catalase, total antioxidant capacity (TAC), and lactate dehydrogenase decreased; ATP secretion, activation and cell viability decreased; and glucose and aggregation increased in CYP compared to CON. Catalase, TAC and glucose were maintained in MEL compared to CON. However, catalase increased, thereby decreasing lipid peroxidation and enhancing activation and aggregation in CYP + MEL compared to CYP. Melatonin mitigates CYP-induced cellular damage by scavenging ROS and augmenting antioxidant defenses. Platelets possess a complex network of signaling cascades, and can serve as ex vivo models to study platelet dysfunction. They can provide insights into platelet behavior under simulated physiological and pathological conditions.
Prostate cancer (PCa) remains a leading cause of cancer-related mortality in aging male, with resistance to enzalutamide (Enza) representing a major therapeutic challenge. This study identifies SLC3A1 as a key driver of Enza resistance. Through integrative bioinformatics analysis of GEO datasets, we found SLC3A1 significantly upregulated in Enza-resistant PCa models. WGCNA and single-cell RNA sequencing confirmed its association with adverse prognosis and activation of key survival pathways. Functional experiments demonstrated that SLC3A1 knockdown reduced AKT phosphorylation, and inhibited colony formation in LNCaP cells. Conversely, SLC3A1 overexpression activated PI3K-AKT signaling, suppressed ferroptosis and conferred Enza resistance in vitro and in vivo. These results reveal SLC3A1 is a promising therapeutic target for advanced prostate cancer.
The accumulation of amyloid beta (Aβ) and tau tangles in the brain leads to Alzheimer's disease (AD). Fisetin, a natural flavonoid, is an antioxidant molecule, and its neuroprotective effects are not clearly understood. Therefore, attempts have been made to evaluate the neuroprotective effects of fisetin using in silico methods and an Aβ1-42-induced neurotoxicity model in human neuroblastoma SH-SY5Y cells. In silico studies demonstrated that fisetin binds strongly and with high stability to different proteins, such as ULK1 (autophagy marker), p21 (senescence/cell cycle marker), and synaptophysin (synaptic marker), which are responsible for maintaining brain health and are implicated in AD. Moreover, Aβ1-42 was also found to bind to these protein targets, indicating that Aβ1-42 and fisetin both target common binding sites. In vitro studies on SH-SY5Y cells further confirmed that fisetin promotes cell survival under the toxic effects of Aβ1-42. It reduced oxidative stress and restored the activities of ion channels, which were impaired by Aβ1-42 treatment. Fisetin increased antioxidant defense and restored the activity of molecules that control brain signals. Overall, fisetin acts on multiple targets to protect neurons by reducing oxidative damage, supporting ion channel activity, and inducing the autophagy process.
The present study focuses on the design, synthesis, characterization, and biological evaluation of two new series of derivatives 4-(4-amino-3-fluorophenoxy)-N-methylpicolinamide (3a-e) and 2-aminobenzothiazole (5a-e) as potential antioxidant and anticancer agents. The compounds were synthesized via a one-pot condensation reaction of 4-(4-amino-3-fluorophenoxy)-N-methylpicolinamide and 2-aminobenzothiazole with various phenyl isocyanates, isothiocyanates, and sulfonamides in the presence of triethylamine at 60°C. Reaction progress was monitored by TLC, and products were purified using column chromatography. Structural elucidation was achieved using FT-IR, 1H NMR, 13C NMR, HRMS, and CHN analyses. Antioxidant activity, evaluated by DPPH and ABTS assays, revealed that compounds 3d and 5b exhibited the highest radical scavenging potential, with IC50 values of 21.45 μg/mL and 20.05 μg/mL (DPPH) and 22.24 μg/mL and 21.98 μg/mL (ABTS), comparable to ascorbic acid. Cytotoxicity assessment by MTT assay demonstrated strong antiproliferative activity of 3d and 5b against MDA-MB-231 breast cancer cells (IC50 = 22.13 μg/mL and 20.11 μg/mL), with minimal toxicity toward normal 3T3-L1 cells. Fluorescence microscopy and flow cytometry confirmed apoptosis profile induction and G2/M phase arrest. The qRT-PCR analysis showed significant down-regulation of Bcl-2, CDK2, and ERK2 gene expression, correlating with apoptotic activation. Molecular docking and MM-GBSA analyses revealed stable and high-affinity binding of 3d and 5b with Bcl-2, CDK2, and ERK2 proteins, supported by multiple hydrogen bonding and π-π interactions. Molecular dynamics simulations (150 ns) confirmed the structural stability of these complexes. Overall, compounds 3d and 5b emerged as promising dual-function 'antioxidant and anticancer' candidates and in vivo evaluations for therapeutic development.
