Search Results (1,610)

The present investigation evaluated the therapeutic potential of ethanol-derived extracts from Kadsura coccinea root (KCR) against alcohol-induced liver injury (ALI) utilizing a murine experimental system. Male Balb/c mice were administered alcohol intragastrically in a stepwise manner over 8 weeks to establish the ALI model. Experimental outcomes demonstrated that KCR administration substantially improved hepatic functional status, evidenced by marked reductions in circulating hepatic enzymes, specifically aspartate aminotransferase (AST) and alanine aminotransferase (ALT). KCR also increased glutathione (GSH) activity, reduced malondialdehyde (MDA) levels in the liver, and exerted antioxidant effects by boosting the expression of enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase 4 (GPX4). Additionally, metabolomic and transcriptomic analyses identified metabolites and pathways closely linked to oxidative stress, including Glutathione metabolism and the MAPK signaling pathway. Further mechanistic studies revealed that KCR could decrease the phosphorylation of p38, JNK, and ERK, while increasing the expression of Nrf2, HO-1, and NQO1. In conclusion, KCR alleviates ALI by modulating the MAPK/Nrf2 pathway, restoring redox homeostasis, enhancing antioxidant defenses, and improving metabolic disorders. Full article

Background/Objectives: Skeletal muscle adaptation to metabolic stress involves a coordinated regulation of inflammatory, bioenergetic, and anabolic signalling pathways. This study aimed to investigate the potential role of whey protein isolate (WPI; commercial name: Volapure) as a modulator of cellular responses to stress in an in vitro model of exercise-mimetic stress over time. Methods: Murine C2C12-differentiated cells were exposed to an Exercise–Mimetic Mix (ExM) to reproduce key biochemical features of muscle stress. Cells were treated with WPI (1 mg/mL) using Pre-exposure (Pre-ExM) and Post-exposure (Post-ExM) protocols at 8 and 24 h. Multiple endpoints were assessed, including cell viability, reactive oxygen species (ROS) production, cytokine release (TNF-α, IL-6, IL-17), intracellular signalling pathways (p38 MAPK, ERK, AMPK, mTOR), bioenergetic markers (ATP, glycogen, lactate), protein synthesis (OPP incorporation), and Ca²⁺/Mg²⁺ fluxes. Results: ExM exposure induced a stress phenotype characterised by increased oxidative and inflammatory markers, impaired bioenergetic status, and reduced anabolic signalling. WPI was associated with modulation of these responses, reducing ROS and pro-inflammatory cytokines, restoring ATP and glycogen levels, and changes in ERK and mTOR-related signalling. The Post-ExM protocol showed greater modulation compared to the Pre-ExM approach, particularly at 24 h. WPI was also associated with the normalisation of ExM-altered Ca²⁺/Mg²⁺ fluxes. These findings should be interpreted as associative rather than causal. Conclusions: WPI was associated with modulation of key pathways involved in cellular adaptation to metabolic stress, supporting recovery of bioenergetic balance and anabolic signalling in C2C12 cells. These findings suggest a potential role for WPI in influencing cellular responses to metabolic stress, supporting recovery of bioenergetic balance and anabolic signalling in C2C12-differentiated-cells. However, further studies are required to confirm the translational relevance of these observations. Full article

Hydrogels constructed from peptide components often rely on β-sheet architectures for their assembly, yet the process of developing such materials in aqueous environments presents notable hurdles in the context of biological systems. To address this, a novel functional dodecapeptide has been developed, capable of self-assembling into supra-molecular hydrogels via zinc chelation interactions. Morphological observations revealed a compact meshwork structure in the hydrogel formed with 9 mM Zn²⁺, differing from the relatively sparse or excessively tangled fiber architectures seen at other zinc concentrations. Alkaline phosphatase activity, an early marker of osteoblast differentiation, was notably enhanced when MC3T3-E1 cells were cultivated for 72 h in the hydrogel extract containing 300 μg/mL of the peptide, 9 μg/mL ZnCl₂, and 18.93 μg/mL H₃BO₃. Furthermore, increased protein levels of p-p38/p38, p-ERK/ERK, and p-JNK1/2/3/JNK1/2/3 were observed in P-300-ZnB and P-300 B hydrogel-treated groups, suggesting an association with MAPK pathway activation. P-Zn-9 hydrogel also promoted MC3T3-E1 cell proliferation and demonstrated favorable biocompatibility in short-term in vitro and in vivo assays. Long-term toxicity and causal relationships via inhibitor studies remain to be investigated. These results offer a viable approach to endow zinc-chelating properties in the fabrication of assembled hydrogels, presenting an innovative and potential method for constructing injectable drug delivery systems and in situ bone repair through biomaterials in subsequent applications. Full article

In peripheral nervous system (PNS), immature Schwann cells differentiate into two functionally distinct types of mature Schwann cells: myelinating Schwann cells, which establish a one-to-one relationship with large-diameter axons and initiate the complex myelination program that enables the rapid saltatory conduction of nerve impulses, and non-myelinating Schwann cells, which envelop multiple small-diameter axons without forming myelin, but support axon survival and maintain microenvironment homeostasis. Here, we demonstrate that discoidin domain receptor 1 (DDR1) signaling plays a pivotal role in Schwann cell maturation and peripheral nerve myelination. Ddr1^−/− mice of both sexes exhibited fewer myelinating Schwann cells and profound hypomyelination, reduced nerve conduction velocity, and apparent motor dysfunction. Expression of myelin markers was markedly reduced in the mutants accompanied by the formation of abnormal myelin ultrastructure resembling “onion bulbs”. Mechanistically, Ddr1 deficiency impaired Schwann cell myelination through the marked hyperactivation of the MAPK/ERK signaling pathway. These findings establish DDR1 as a novel regulator of Schwann cell myelination and highlight its potential as a therapeutic target for demyelinating neuropathies. Full article

Diabetic nephropathy (DN) is a leading cause of end-stage renal disease with limited therapeutic options. Ferroptosis contributes to renal tubular injury in DN. This study investigates whether mesenchymal stem cells (MSCs) ameliorate DN by inhibiting ferroptosis and elucidates the underlying mechanism. In a rat model of type 2 DN, MSCs transplantation improved renal function and histopathology, while reducing mitochondrial dysfunction, iron overload, and ROS-driven ferroptosis. In vitro, MSCs reversed high glucose-induced ferroptosis hallmarks in tubular epithelial cells. Mechanistically, RNA sequencing identified the MAPK/ERK pathway as key. MSCs suppressed the p-ERK/ERK-GPX4/ACSL4 axis, preventing glutathione depletion and lipid peroxidation. Activation of ERK abolished MSCs’ protection, whereas ERK inhibition mimicked it. These findings reveal that targeting ERK-mediated ferroptosis in renal tubules offers a novel therapeutic strategy, with MSCs acting through this specific mechanism. Full article

Cucurbitacin B belongs to a group of tetracyclic triterpenoids and exerts a number of biological effects, including anti-inflammatory and anticancer activities. We previously demonstrated that cucurbitacin B down-regulated tumor necrosis factor (TNF) receptor 1 (TNF-R1) expression and prevented activation of the transcription factor nuclear factor κB in response to a TNF-α stimulation. The present study shows that cucurbitacin B promoted the ectodomain shedding of TNF-R1 by generating a soluble form that accumulated in the culture medium of human lung adenocarcinoma A549 cells. Of the eight tetracyclic and pentacyclic triterpenoids consisting of an α,β-unsaturated carbonyl group that were examined, only cucurbitacin B promoted TNF-R1 ectodomain shedding. Cucurbitacin B-induced TNF-R1 shedding was attenuated by TNF-α protease inhibitor 2 and the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor U0126, but not by the p38 MAPK inhibitor SB203580 or the c-Jun N-terminal kinase (JNK) inhibitor SP600125. Consistent with these results, cucurbitacin B promoted the rapid phosphorylation of rapidly accelerated fibrosarcoma 1 (RAF1) and ERK, but exerted minimal effects on the phosphorylation of p38 MAPK and JNK. Collectively, these results demonstrate that cucurbitacin B selectively activated the RAF1-MEK-ERK pathway, which was essential for TNF-R1 ectodomain shedding. Full article

Dental composite dust generated during finishing procedures or mastication may adversely affect gingival epithelia. However, the mechanistic distinction between particulate and chemical (eluate) exposures and their respective signaling consequences remains insufficiently defined. Dust particles and corresponding eluates from three restorative composites, Admira Fusion, Ceram.x Spectra ST, and Filtek Supreme XTE, were evaluated under standardized high-dose in vitro exposure conditions. Human gingival keratinocytes were assessed for proliferation, adhesion, differentiation, fibronectin (FN1) remodeling, and IL-8 secretion, alongside analysis of ERK, p38, and NF-κB signaling and phosphorylation of the stress-responsive regulator SIRT1 at Ser682 (SIRT1-S682). Particulate exposure elicited more pronounced impairment of cellular adhesion, proliferation, and differentiation than eluates. Dusts derived from Ceram.x Spectra ST and Filtek Supreme XTE suppressed ERK activity, reduced FN1 abundance, and decreased nuclear SIRT1-S682, consistent with a generalized stress response. In contrast, Admira Fusion dust preserved FN1, activated ERK signaling, reduced SIRT1-S682, and induced robust IL-8 secretion. Across all materials, particulate exposure reduced nuclear SIRT1-S682 without affecting total SIRT1 levels, indicating a shared permissive stress modification. Notably, only Admira Fusion coupled this permissive state with p38 activation and sustained NF-κB p65 Ser536 phosphorylation, resulting in transcriptionally active NF-κB and elevated IL-8 production, whereas Ceram.x Spectra ST and Filtek Supreme XTE failed to activate this ERK–FN1–p38–NF-κB axis, yielding either transcriptionally inactive NF-κB or no detectable enrichment. These findings support a material-associated in vitro response pattern in which a shared SIRT1-S682 reduction is accompanied by distinct ERK/FN1, p38, NF-κB, and IL-8 readouts. SIRT1-S682 reduction alone did not define the inflammatory phenotype, because it occurred across particulate exposures, whereas IL-8 secretion was observed only under conditions that also showed p38 activation and comparatively maintained NF-κB p65 Ser536 phosphorylation. This signature arises from the convergence of a permissive SIRT1-S682 background with ERK- and p38-dependent MAPK signaling to enable NF-κB-mediated IL-8 expression, highlighting that both composite composition and particulate properties critically determine inflammatory potential and underscoring the importance of incorporating particulate fractions into cytocompatibility testing strategies. Full article

Background/Objectives: PI3K/AKT/mTOR is a key pathway in cell proliferation, metabolism, and survival. Activating PIK3CA mutations are seen in up to 20% of colorectal cancers and are associated with increased cyclo-oxygenase-2 (COX-2) expression. Recent studies demonstrated a significant survival benefit from taking low-dose aspirin, a nonselective COX inhibitor, supporting further exploration of the synergistic effects of combined PI3Kα inhibitor (inavolisib) and COX-2 inhibitor (celecoxib) therapy. Methods: The effects of celecoxib–inavolisib combination treatment were tested on human colorectal cancer cell lines and patient-derived organoid models. Experiments included cell viability and colony formation assays, immunoblotting, and immunofluorescence. Results: We found that celecoxib and inavolisib demonstrated synergy in suppressing the growth of colorectal cancer cell lines, grown in both 2D and 3D cell culture, regardless of PIK3CA mutation status. In patient-derived organoid models, while synergy was seen in both organoids, growth of the PIK3CA mutated organoid was more potently suppressed. Immunoblotting of cells after combination treatment showed decreased expression of mitogenic signaling marker p-AKT across all 2D cell lines and in both cell lines grown as 3D spheroids, as well as increased expression of apoptotic marker cPARP in four out of five 2D cell lines and in both cell lines grown as 3D spheroids. Immunofluorescence staining of organoids after combination treatment, however, showed no significant increase in expression of apoptotic marker Cas-3 nor in mitogenic marker Ki-67 in either organoid. Furthermore, an apoptosis assay performed on two cell lines showed no significant increase in Annexin V or phosphatidylserine staining. Conclusions: Celecoxib and inavolisib demonstrated synergy in suppressing the growth of both colorectal cancer cell lines and patient-derived organoids, though PIK3CA mutation status did not appear to affect drug efficacy in cell lines as it did in patient-derived organoids. Potential compensatory or resistance mechanisms might include oncogene drivers in the MAPK/ERK pathway. When compared to monotherapy, combination therapy was the only drug condition to significantly increase the percentage of apoptotic cells based on Annexin V and phosphatidylserine staining, and this effect was only seen in the PIK3CA mutated cell line. Ultimately, our findings provide preliminary support for celecoxib–inavolisib combination treatment as a rational therapeutic avenue warranting further preclinical investigation. Full article

Background: Endometrial cancer is a common gynecological malignancy, and elucidating its molecular basis may provide new clues for targeted intervention. This study investigated the role of ERBB3 in endometrial cancer cells and explored whether Ras-ERK/MAPK signaling is involved in ERBB3-mediated regulation. Methods: Bioinformatic screening was performed using public databases to identify candidate genes associated with endometrial cancer. ERBB3 was selected for further analysis. ERBB3 expression was evaluated in public datasets and examined in endometrial cancer cells. Loss-of-function experiments, rescue assays, Western blotting, and co-immunoprecipitation were used to assess the biological function and potential mechanism of ERBB3. Results: ERBB3 knockdown significantly inhibited proliferation, migration, and invasion, and promoted apoptosis in Ishikawa and RL95-2 cells. These changes were accompanied by decreased Ras-ERK/MAPK signaling. Moreover, Ras overexpression partially reversed the effects induced by ERBB3 knockdown. Co-immunoprecipitation suggested a molecular association between ERBB3 and Ras within a protein complex, but did not demonstrate direct physical binding. Conclusions: ERBB3 appears to promote malignant behaviors in endometrial cancer cells, and the Ras-ERK/MAPK pathway may be one of the downstream mechanisms involved. Further validation in additional experimental models and clinical samples is needed. Full article

Background: The retinal pigment epithelium (RPE) plays a pivotal role in the visual process by maintaining the blood–retina barrier, protecting the retina from oxidative stress, and regulating immune responses. Consequently, dysfunction or degeneration of the RPE is implicated in a broad spectrum of retinal disorders that lead to progressive and irreversible vision loss. In this context, inflammation of the RPE has emerged as a critical factor in the pathogenesis of retinal degenerative diseases, underscoring its dual role as both a target and mediator of retinal inflammatory processes within the retina. Objectives: This study aims to preliminarily investigate, mainly by assessment of proinflammatory cytokine gene expression and immunoblotting, the molecular mechanisms underlying RPE inflammation induced by interactions between the RPE and microglia of the central nervous system. Methods/Results: Using in vitro models of human RPE cells, the ARPE 19 cell line was exposed to conditioned media from microglia (CHME-5 cell line) under basal and proinflammatory conditions. We observed increased activation of the MAPK signaling pathway, (evidenced by a 4-fold increase in the phosphorylation ratio of MEK and ERK) alongside elevated expression of proinflammatory cytokines, assessed by RT-PCR and immunoblotting, and a 2-fold increase in reactive oxygen species levels in RPE cells, evaluated by colorimetric assays, after exposure with conditioned media. Specifically, IL-1β and IL-8 levels increased more than 40-fold, while IL-6 expression showed a 4-fold increase compared to controls. Conclusions: These findings emphasize the central role of the RPE in retinal inflammation and suggest potential therapeutic targets to modulate immune responses and preserve retinal function. Full article

Computer-aided drug discovery increasingly depends on virtual-screening workflows that remain reliable under severe class imbalance, chemical redundancy and early-recognition constraints. In this study, we developed a leakage-aware prioritization workflow for two cancer-relevant targets, pyruvate kinase M2 (PKM2) and mitogen-activated protein kinase 1 (MAPK1/ERK2), using the LIT-PCBA benchmark. The workflow combines canonical-SMILES curation, duplicate and label-conflict auditing, scaffold-aware validation, a non-learning nearest-active Tanimoto baseline, imbalance-aware machine-learning models, repeated-seed robustness analysis, isotonic probability calibration, ensemble-disagreement estimation, absorption, distribution, metabolism, excretion and toxicity (ADMET)-aware triage, molecular docking, and residue-level contact analysis. Benchmark enrichment is interpreted alongside calibration, ADMET filtering, docking and residue-contact evidence, rather than as a standalone discovery claim. PKM2 emerged as the clearer machine-learning case, with scaffold-aware tree models improving early recognition beyond the nearest-active similarity baseline and yielding top-ranked candidates supported by calibrated activity scores, ADMET profiles, docking scores, and residue-contact fingerprints. MAPK1 provided a biologically relevant contrast target, where ligand-neighborhood similarity remained competitive and downstream structural triage became more decisive than ligand-based ranking alone. These results support a conservative drug-discovery workflow in which leakage-aware benchmarking, calibration, uncertainty, and molecular-level triage remain visible throughout candidate prioritization. Full article

Ischemic stroke is an acute cerebrovascular disease with high disability and morbidity. However, therapeutic approaches are restricted by a narrow time window for reperfusion. Astilbin has various pharmacological activities and good therapeutic potential against ischemic stroke and neurodegenerative diseases. Nevertheless, Astilbin’s mechanism of action remains unclear. Here, we used an integrated strategy that includes network pharmacology, omics validation, and functional verification. Potential targets of Astilbin were predicted using SwissTargetPrediction and PharmMapper, and cross-analyzed with IS-related genes from multiple databases. GO/KEGG enrichment analyses showed that Astilbin synergistically regulates stroke-associated pathways (e.g., MAPK, AGE-RAGE). Combined transcriptomic and metabolomic assays confirmed that Astilbin ameliorates OGD/R-induced oxidative stress and metabolic disorders by modulating the MAPK and ferroptosis pathways. Molecular docking and dynamics simulations revealed that Astilbin has high affinity for core targets (ERK1/2, CREB, p90RSK, MMP9) and binds stably to MMP9. Using an OGD/R-injured neuronal-like PC12 cell line, in vitro assays confirmed that Astilbin alleviates oxidative stress, calcium overload, lipid peroxidation, and intracellular iron levels, while also modulating apoptosis- and inflammation-related genes. Overall, this study has established a comprehensive pharmacological framework for the use of Astilbin against IS, clarified its multi-target, multi-pathway neuroprotective mechanisms of action, and provided evidence for its potential in the treatment of IS. Full article

Pancreatic tumors frequently exhibit calcification, suggesting potential osteogenic-related phenotypic plasticity. This study aimed to systematically evaluate whether pancreatic ductal adenocarcinoma (PDAC) cells acquire osteogenic-like features under induction conditions and to assess the associated phenotypic and molecular changes. PDAC cell lines and non-malignant pancreatic epithelial cells were subjected to osteogenic induction. Mineralization, alkaline phosphatase (ALP) activity, osteogenic marker expression, and malignant phenotypes were evaluated. RNA sequencing was performed at defined time points to characterize transcriptional changes. Pharmacological inhibition of MEK and siRNA-mediated knockdown of RUNX2 were applied to examine the involvement of MAPK–ERK signaling and downstream transcriptional regulation. Osteogenic induction led to calcium deposition and increased ALP activity in a subset of PDAC cell lines, accompanied by upregulation of osteogenic-associated markers, including RUNX2 and SPP1. Induced cells exhibited reduced migration, clonogenicity, invasion, and proliferation. Transcriptomic analysis revealed activation of osteogenesis-related and calcium-transport pathways, along with downregulation of cell cycle programs. MAPK–ERK signaling was activated during induction, and MEK inhibition attenuated RUNX2 and ALP expression as well as mineralization-associated changes. Furthermore, RUNX2 knockdown reduced ALP expression and mineralization levels, indicating its contribution to the osteogenic-like phenotype. PDAC cells can acquire osteogenic-like features under defined induction conditions, accompanied by coordinated transcriptional reprogramming and reduced malignant phenotypes. The observed mineralization-associated phenotypes may reflect a combination of active processes and passive calcium deposition. In addition, the MAPK–ERK–RUNX2 axis appears to be involved in this process, although it may reflect a broader adaptive or stress-associated reprogramming rather than lineage commitment. These findings provide insight into the potential relationship between tumor calcification and phenotypic plasticity in PDAC. Full article

Background/Objectives: Salvia miltiorrhiza is a medicinal plant rich in phenolic acids and tanshinones, compounds that have been linked to anti-inflammatory and neuroprotective activities. ‘Hongdan’ is a Korean cultivar characterized by relatively high levels of salvianolic acid B and tanshinone IIA, but its anti-inflammatory activity in microglial cells has not yet been examined. Methods: Nitrite production and the mRNA expression of inflammatory mediators (iNOS and COX-2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) were examined. In addition, activation of MAPK (ERK1/2, JNK, and p38) signaling pathway and expression of the NF-κB regulatory protein IκB-α were analyzed. Results: The Hongdan extract inhibited nitrite production and reduced the expression of iNOS and COX-2 in LPS-stimulated BV2 microglial cells. In addition, the expression of IL-1β and IL-6 was markedly reduced, whereas TNF-α was significantly suppressed only at the highest concentration tested. Furthermore, phosphorylation of ERK1/2, JNK, and p38 was significantly inhibited, while IκB-α degradation was not altered. Conclusions: These findings demonstrate that the Hongdan extract effectively suppresses LPS-induced inflammatory responses through inhibition of MAPK signaling pathways and may serve as a promising natural therapeutic candidate for neuroinflammatory disorders. Full article

Glycosylation is essential for hematopoietic cell homeostasis and malignant transformation. Dysregulated expression of glycosylation genes in leukemia cells accelerates disease progression and fosters drug resistance. Therefore, targeting these genes offers a promising avenue for anti-leukemic therapy. In this study, we explore the roles of N-glycans in acute promyelocytic leukemia (APL) differentiation using the ATRA-induced wild-type NB4 (WT/ATRA) or HL-60 cell model. We found that expression of N-acetylglucosaminyltransferase IVa (GnT-IVa, encoded by the MGAT4A gene) and its product (β1,4-GlcNAc-branched N-glycan) increased significantly during differentiation, as evaluated by lectin blot, real-time PCR, and flow cytometry. Interestingly, analysis of the Gene Expression Omnibus (GEO) public data showed that MGAT4A expression is significantly lower in APL patients, and higher MGAT4A expression was associated with favorable survival in AML cohorts. To address the role of GnT-IVa in differentiation, we established MGAT4A- and MGAT4B-knockout (KO) NB4 cell lines using CRISPR/Cas9. Compared to WT/ATRA cells, MGAT4A KO, but not MGAT4B KO, markedly suppressed ATRA-induced differentiation, as evidenced by reduced expression of CD11b and CD11c. We found that CD11b is a major glycoprotein carrying β1,4-GlcNAc-branched N-glycans. This modification enhanced CD11b stability, as CD11b expression declined more rapidly in MGAT4A KO cells in the presence of cycloheximide. In addition, MGAT4A KO suppressed ERK/MAPK signaling, which contributed to differentiation. Our study highlights the critical role of GnT-IVa in regulating APL differentiation, which may provide a basis for developing new differentiation therapies for APL. Full article