Molecular Biology Reports最新文献

Background: Oxidative stress, apoptosis, and inflammation are interconnected pathological processes. Bryophytes, such as Dicranum scoparium, have evolved unique secondary metabolic systems, including potent antioxidants, to survive extreme environmental stressors such as dedication and UV radiation. This study aimed to characterize the chemical composition of D. scoparium aqueous extract (DSAE) and evaluate its multi-targeted therapeutic potential using an integrated metabolomics and in vitro co-culture approach.

Methods: The chemical fingerprint of DSAE was established via GC-MS using a DB-5MS column (split ratio 1:10). Cytoprotective effects were evaluated in NIH/3T3 fibroblasts across a 100-fold serial concentration range (0.027 ng/mL, 2.7 ng/mL, 270 ng/mL, 2.7 µg/mL, and 27 µg/mL). A fibroblast-macrophage co-culture model was employed to monitor intercellular signaling and the regulation of the NOX2/SOD2 and NF-κB/inflammasome pathways under H₂O₂-induced stress.

Results: GC-MS identified 34 metabolites, including erythritol and GABA-related derivatives. DSAE (0.027 ng/mL) significantly restored cell viability (88.4 ± 4.1% vs. 50.2 ± 3.4% in H₂O₂) and reduced ROS mean fluorescence intensity by 44.6 ± 5.2%. DSAE significantly upregulated HO-1 (1.25 ± 0.14-fold, p < 0.01) and GPx1 (0.88 ± 0.09-fold, p < 0.05). In the co-culture system, DSAE accelerated wound closure speed (3.44 ± 0.26 μm/h) and suppressed the transcription of NLRP3 (1.84 ± 0.21-fold) and IL-6 (1.92 ± 0.25-fold) via NF-κB inhibition.

Conclusions: DSAE exerts multi-targeted cytoprotective effects by modulating the NOX2/SOD2 axis and suppressing intercellular inflammatory crosstalk. These findings suggest that D. scoparium is a valuable source of bioactive metabolites for treating chronic inflammatory and oxidative disorders.

Background: Hepatocellular carcinoma (HCC) remains a major global health burden, partly due to the lack of physiologically relevant in vitro models that accurately recapitulate early host-virus interactions and immune responses. Human Hepatocyte Line 5 (HHL-5) is an immortalized hepatocyte cell line that retains key liver-specific functions. This study aimed to characterize the phenotypic, genetic, and metabolic features of HHL-5 cells and evaluate their suitability as a non-cancerous hepatic model, in comparison with the HCC cell line HepG2.

Methods and results: Morphological and phenotypic assessment of cells showed smaller cell and nuclear areas and slower proliferation with markedly longer doubling time of HHL-5 cells than HepG2 cells. Genomic analyses using whole-exome sequencing revealed enrichment of immune-related pathways in HHL-5 cells, including antigen processing and presentation, whereas HepG2 cells showed predominance of DNA replication pathways. Metabolomic profiling of cells by nuclear magnetic resonance spectroscopy showed hepatocyte-like oxidative profiles of HHL-5 cells, in contrast to the glycolytic phenotypes of HepG2 cells. Moreover, Western blotting for selected proteins showed reduced expression of oncogenic and stress-response markers, including c-Myc, pSTAT3, pNrf2, and select cytochrome P450 enzymes.

Conclusion: Our findings support HHL-5 cells as a robust non-cancerous in vitro model for investigating liver diseases, viral infection, and early events in hepatocarcinogenesis.

Clinical trial registration: Not applicable.

Background: Biologic therapies targeting TNF-α in inflammatory and autoimmune diseases underscore its central role in pathological inflammation. Epigenetic mechanisms are increasingly recognized as critical modulators of disease-associated gene expression, and advancing epigenetic-based therapeutics requires deeper insight into the transcriptional networks governing inflammatory responses. Here, we investigate the epigenetic regulation of inflammation-induced TNF-α transcription and identify lead compounds that suppress TNF-α expression in human macrophages.

Methods and results: A small-molecule epigenetic inhibitor screen comprising 152 compounds was performed to assess their effects on LPS-induced TNF-α transcription in THP-1 monocyte-derived macrophages. TNF-α mRNA and protein levels were quantified using qRT-PCR and ELISA, respectively. The screen identified 20 potent inhibitors of TNF-α expression, representing six major classes of epigenetic regulators: HATs, BRDs, HDACs, HDMs, PRMT1, and PLK1. Protein-protein interaction network analysis revealed extensive crosstalk between these targets and key inflammatory signaling pathways, including NF-κB and MAPK. Follow-up validation of the top 10 candidates demonstrated dose-dependent suppression of TNF-α at both transcriptional and protein levels. These compounds also exhibited broad anti-inflammatory activity, suppressing IL-1β, IL-6, IL-8, IL-17A, and CCL2 expression, and showed cross-species efficacy in human (THP-1) and mouse (RAW264.7) macrophages. NF-κB reporter assays further indicated that their inhibitory effects are mediated, at least in part, through blockade of NF-κB activation.

Conclusions: This study identifies a regulatory network of epigenetic modifiers controlling TNF-α transcription and reveals conserved mechanisms governing inflammatory gene expression. The validated lead compounds possess potent anti-inflammatory properties and hold strong potential for therapeutic repurposing as modulators of TNF-α-driven inflammatory diseases.

Background: Thyroid hormones are mainly concerning with regulation of organs metabolism. Hypothyroidism is a pathological condition characterized by decrease of T3 and T4 level that negatively impact various organs especially liver and lung. Dapagliflozin (DAPA) is an anti diabetic drug posses antioxidant and anti apoptotic properties AIM OF THE STUDY: The aim of this work is to identify the effect of DAPA on disturbance of liver and lung functions result from hypothyroidism MATERIALS AND METHODS: Twenty four rats (6 rats/group) were divided into four groups: control group, DAPA group that received DAPA (1 mg/kg/day for five weeks), PTU group that received propylthiouracil (15 mg/kg/day for five weeks), PTU+ DAPA group that received PTU and DAPA as described in the other groups RESULTS: Hypothyroidism induced oxidative stress associated mitophagy disturbance in both lung and liver with development of apoptosis in liver. DAPA avoided this effect CONCLUSION: DAPA protects against liver and lung injury result from hypothyroidism.

Dimethyl fumarate (DMF) is an electrophilic compound used clinically for multiple sclerosis and psoriasis. We elaborate here that the pharmacological effects of DMF extend beyond the well-known activation of the NRF2 antioxidative pathway. Indeed, DMF directly modifies reactive cysteine residues on multiple proteins in immune and neural cells, leading to diverse anti-inflammatory, immunomodulatory, and neuroprotective actions. Recent studies revealed that DMF may affect proteins involved in inflammasome activation, glycolysis, and cell signaling pathways, including JAK-STAT and NF-kB. These effects may expand the potential clinical applications of DMF in diverse pathologies, including neurodegenerative, cardiovascular, and pulmonary diseases. We summarize current findings on chemical reactivity, target proteins, and emerging clinical applications of DMF, highlighting new opportunities for DMF repurposing.

Background: ADAMTS-2 is a key extracellular matrix (ECM) remodeling enzyme increasingly implicated in osteosarcoma biology. Although ADAMTS-2 is known to participate in collagen processing and ECM turnover, the upstream inflammatory cues and transcriptional mechanisms regulating its expression in osteosarcoma remain unclear. Tumor necrosis factor-α (TNF-α), a dominant pro-inflammatory cytokine within the osteosarcoma microenvironment, represents a strong candidate regulator due to its ability to activate several pathways. This study aimed to elucidate the TNF-α-ADAMTS-2 regulatory axis at mechanistic levels.

Methods and results: Gene expression profiling revealed that ADAMTS-2 is significantly upregulated in osteosarcoma tissues compared with normal bone and is associated with ECM disassembly and collagen fibril organization pathways. Functional assays in Saos-2 cells demonstrated that TNF-α stimulation markedly increased ADAMTS-2 mRNA (~ 17-fold) and protein levels (~ twofold). Promoter activity assays showed strong TNF-α-mediated activation, with the highest induction in the - 180/ + 112 region. Pharmacological inhibition experiments revealed that MEK, PI3K, JNK, and NF-κB pathways are all required for TNF-α-induced ADAMTS-2 transcription. In silico motif analysis identified STAT3 and NF-κB binding elements within the promoter, and electrophoretic mobility shift assays supported the interaction of these transcription factors with specific promoter regions.

Conclusions: This study provides the first mechanistic evidence that TNF-α regulates ADAMTS-2 expression through the coordinated activation of multiple signaling pathways and the engagement of STAT3 and NF-κB transcription factors at the promoter. These findings uncover a previously uncharacterized inflammatory regulatory axis linking TNF-α signaling to ECM remodeling and highlight ADAMTS-2 as a potential mediator of osteosarcoma progression.