Cellular Physiology and Biochemistry最新文献

Background/aims: To explore the feasibility and effectiveness of Tumor Destructive Mechanical Impulse (TMI) treatment of solid tumors, biomechanical preconditions for subsequent computational simulation of focused shock wave propagation within cells and tissue are investigated. This innovative "soft" approach is different from the FDA-approved high intensity focused ultrasound (HIFU)-based histotripsy, and from electrical Tumor Treating Fields (TTFs).

Methods: Atomic force microscopy investigation for cell mechanics, multiple parametric computational simulations for focused shock wave propagation, technical TMI generator and applicator design, light- and electron-microscopic evaluation of treatment effects on tumor cells and tissue.

Results: Individual tumor cell evaluation of physical properties as basis for multiple parametric simulations determine the optimal treatment parameters (total energy required, energy flux density, shock wave frequency) and applicator positions; design flexibility of applicator devices for extra- and intracorporeal treatment.

Conclusion: The fundamental feasibility, effectiveness and reliability of TMI treatment of solid tumors were proven, providing a reliable theoretical basis for the broadly applicable translation into clinical practice.

Background/aims: Myocardial infarction remains a leading cause of cardiovascular morbidity and mortality. Although reperfusion therapy restores myocardial blood flow, it can induce myocardial ischemia-reperfusion injury (MI/RI). Ferroptosis, an iron-dependent form of regulated cell death driven by excessive lipid reactive oxygen species, contributes to MI/RI and is characterized by downregulation of GPX4 and upregulation of ACSL4. LncRNA ANRIL is aberrantly expressed in acute myocardial infarction and may provide myocardial protection, but its role in MI/RI-induced ferroptosis is unclear.

Methods: A mouse MI/R model was established by ligating the left anterior descending coronary artery in C57BL/6 mice. HL-1 and H9C2 cardiomyocytes underwent hypoxia-reoxygenation to simulate MI/RI in vitro. Lnc-ANRIL was overexpressed using pEGFP-lnc-ANRIL or silenced using siANRIL. Ferroptosis indicators (ROS, malondialdehyde, Fe2+, GPX4, ACSL4) were assessed. Candidate miRNAs targeting lnc-ANRIL and GPX4 were predicted (miRDB) and validated by dual-luciferase assays.

Results: Ferroptosis was activated in MI/R tissues and hypoxia-reoxygenation-treated cardiomyocytes, with decreased GPX4, increased ACSL4, and elevated ROS, malondialdehyde, and Fe2+. Lnc-ANRIL expression was reduced. Overexpression of lnc-ANRIL attenuated ferroptosis markers and increased GPX4, whereas lnc-ANRIL silencing exacerbated ferroptosis. Mechanistically, lnc-ANRIL acted as a sponge for miR-7238-3p, which targets the 3'-UTR of GPX4 to suppress expression. Overexpression of miR-7238-3p enhanced ferroptosis and cardiomyocyte damage.

Conclusion: MI/RI downregulates lnc-ANRIL, relieving inhibition of miR-7238-3p and suppressing GPX4, thereby triggering ferroptosis in cardiomyocytes. Lnc-ANRIL protects against MI/RI-induced ferroptosis via the miR-7238-3p/GPX4 axis, suggesting a potential therapeutic target.

Background/aims: Acute myeloid leukemia (AML) is a devastating hematological malignancy without a definitive cure. 6-Gingerol, a bioactive compound, has shown promise in treating various cancers, yet its impact on AML remains elusive.

Methods: Cell growth and clonogenic capacity were assessed using CCK-8 testing and colony formation assays. Flow cytometry was employed to analyze cell cycle progression and apoptosis. The invasive capability of AML cells was evaluated through the Transwell migration assay. Fluorescent probe staining was used to determine intracellular reactive oxygen species (ROS) concentration, while Western blot was utilized to assess the expression levels of key proteins including Bcl-2, caspase3, MAPK, and p-MAPK in AML cells. Potential targets of 6-gingerol in AML were identified through bioinformatics databases (STP, SEA, STICH, OMIM GeneMap, GeneCards). GO and KEGG enrichment analysis was performed using clusterProfiler (v4.16.0).

Results: 6-Gingerol inhibited proliferation, colony formation, and invasive capacity of AML cells and induced G1 cell-cycle arrest. 6-Gingerol increased ROS and elevated caspase 3, MAPK, and p-MAPK levels. Sixty-seven overlapping targets between 6-gingerol and AML were identified and enriched in MAPK signaling and ROS-related pathways. NFKB1 emerged as a pivotal hub gene.

Conclusion: 6-Gingerol may represent a promising Traditional Chinese Medicine-derived agent for AML treatment.

Bronchial and vascular architecture in the rat lung forms an interdependent scaffold that balances ventilation with perfusion and adapts to metabolic demand. Development proceeds through coordinated branching programs that couple epithelial growth with vascular patterning while matrix remodeling and epithelial-mesenchymal crosstalk shape airway caliber and capillary alignment. Quantification has moved from classical design-based stereology to organ-scale µCT, optical clearing, and multiscale computational reconstructions that link structure to function. Across disease models, Chronic Obstructive Pulmonary Disease (COPD) and emphysema show distal airspace enlargement with vascular rarefaction, pulmonary hypertension (PH) features medial thickening and arteriolar muscularization, asthma combines epithelial remodeling with angiogenesis, and fibrosis exhibits collagen deposition with capillary regression. Convergent signaling networks integrate these changes, including VEGF and HIF pathways that govern angiogenesis, Notch and Wnt programs that regulate morphogenesis, and oxidative stress with cytokine and microRNA axes that drive vascular remodeling. Translational alignment is strengthened by single-cell and imaging biomarkers that map rat phenotypes to human pathology, while bioengineered platforms and in silico models provide controllable test beds for hypothesis testing. Predictive frameworks for remodeling across development and disease could be provided by standardized pipelines that combine morphometry, mechanics, and molecular profiles.

Background/aims: Essential oils (EOs) derived from conifers of the Pinaceae family are complex bioactive mixtures known for their antioxidant and antimicrobial properties. However, their impact on human redox homeostasis, particularly in blood, remains poorly understood. This study aimed to compare the redox-modulating and membrane-stabilising effects of essential oils from Scots pine (PEO), European spruce (SEO), and European silver fir (FEO) using an in vitro human blood model.

Methods: The chemical composition of each EO was characterised using gas chromatography-mass spectrometry (GC-MS), proton-transfer-reaction mass spectrometry (PTR-MS), and Fourier-transform infrared spectroscopy (FTIR). Human blood samples were incubated with different EO concentrations, and oxidative stress biomarkers, antioxidant enzyme activities, and erythrocyte membrane stability were evaluated.

Results: All EOs exhibited terpene-rich profiles dominated by α-pinene, β-pinene, borneol, and bornyl acetate, with distinct species-specific differences. The oils displayed concentration-dependent biphasic redox effects. At moderate concentrations, PEO and SEO enhanced total antioxidant capacity and increased catalase and ceruloplasmin activities by 15-25% (p < 0.05). In contrast, higher doses-particularly of FEO-induced lipid peroxidation and protein oxidation by 40-60% (p < 0.05), indicating pro-oxidant behaviour. Erythrocyte haemolysis assays revealed that SEO exerted the strongest membrane-stabilising effect (haemolysis reduced by 18%), whereas FEO increased membrane fragility (haemolysis increased by 27%).

Conclusion: Pinaceae-derived essential oils exhibit dual antioxidant and pro-oxidant activity dependent on concentration and species. Among them, PEO showed the most balanced redox profile. These findings highlight both the therapeutic potential and the importance of controlled dosing when considering such oils for biomedical applications.

Background/aims: Emerging evidence suggests that circular RNAs (circRNAs) play a crucial role in kidney disease regulation. However, their functional significance in chronic kidney disease (CKD) remains poorly understood.

Methods: In this study, circRNAs were identified by RNA sequencing in two CKD mouse models, including unilateral ureteral obstruction (UUO) and anti-glomerular basement membrane (anti-GBM) glomerulonephritis. RNase-R treatment and Sanger sequencing were used to confirm circular structure. Circ-RNF216-shRNA was used to establish stable knockdown mTEC cell lines. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect circ-RNF216 expression. In situ hybridization was used to assess the expression and localization of circ-RNF216 in kidney. Transwell assays were performed to assess cell migration. RNA sequencing was performed to characterize mRNA expression profiles and identify pathways affected by circ-RNF216 knockdown in mTECs.

Results: We identified 1,589 circRNAs in two CKD mouse models. Circ-RNF216 expression was up-regulated in UUO models. Functional analyses revealed that circ-RNF216 regulates renal fibrosis through modulation of the TGF-β1/Smad3 pathway. Knocking down circ-RNF216 in mouse tubular epithelial cells led to significant suppression of migratory capacity and fibrosis. RNA sequencing showed that circ-RNF216 knockdown altered mRNA expression, with differentially expressed genes mainly enriched in the transforming growth factor-β receptor superfamily signaling pathway.

Conclusion: Our findings highlight circ-RNF216 as a novel regulatory factor in CKD-related renal fibrosis, broadening our understanding of circRNA involvement in kidney disease pathogenesis and suggesting circ-RNF216 as a potential therapeutic target for preserving renal function.

Background/aims: Succinylation, a recently characterized post-translational modification (PTM), is a ubiquitously occurring protein modification implicated in diverse biological processes via regulation of protein function and gene expression. CTBP1 encodes C-terminal binding proteins and generates multiple splice variants. However, the functional significance of CTBP1 succinylation in hepatocellular carcinoma (HCC) remains unexplored.

Methods: Protein succinylation levels were quantified using immunoprecipitation coupled with Western blotting and mass spectrometry. Site-directed mutagenesis identified critical lysine residues targeted by succinylation. Functional impacts of CTBP1 succinylation on HCC cell behaviors were evaluated through CCK8-based cell viability, wound healing, transwell migration, and invasion assays. Molecular mechanisms were elucidated via qRT-PCR and Western blot analyses.

Results: Succinylation levels of CTBP1 were significantly elevated in HCC tumor tissues and cell lines relative to non-tumorous controls. Mass spectrometry and mutagenesis pinpointed K46 and K280 as the primary succinylation sites on CTBP1, with SIRT5 identified as the desuccinylase. Functionally, CTBP1 succinylation enhanced HCC cell proliferation, migration, and invasive potential. Mechanistically, this modification promoted tumor progression by suppressing MAT1A expression-a key regulator of hepatic differentiation and tumorigenesis.

Conclusion: Our study reveals that SIRT5-mediated CTBP1 succinylation drives HCC progression through MAT1A suppression, establishing a novel regulatory axis with therapeutic potential for HCC treatment.

Background/aims: Adipogenesis involves preadipocyte differentiation and lipid droplet accumulation and is closely linked to obesity. 3-Acetyl-11-keto-beta-boswellic acid (AKBA), a frankincense-derived triterpene, has anti-inflammatory and anticancer properties, but its role in adipocyte differentiation remains unclear.

Methods: 3T3-L1 preadipocytes were induced to differentiate with or without varying concentrations of AKBA. Cell viability was assessed by MTT assay, lipid accumulation by Oil Red O staining, and apoptosis by annexin V-FITC assay. Expression of adipogenic transcription factors, lipid-associated proteins, apoptotic markers, autophagy-related proteins, and AMPK phosphorylation was analysed by Western blotting. Molecular docking was used to evaluate AKBA interactions with target proteins.

Results: AKBA inhibited adipocyte differentiation by suppressing C/EBPβ, C/EBPα, and PPARγ expression, reduced lipid accumulation, increased apoptosis via an elevated Bax/Bcl2 ratio, suppressed autophagy markers ATG5 and LC3b, and enhanced AMPK phosphorylation. Docking studies suggested AKBA binding to PPARγ and ATG5.

Conclusion: AKBA suppresses adipogenesis by inhibiting adipogenic transcription factors, autophagy, and activating AMPK, supporting its potential as an anti-obesity therapeutic candidate.

Background/aims: This study aimed to clarify the optimal amount of vitamin E required in the late stage of egg laying to explore the aging mechanism of the ovaries in this stage and the regulatory role of vitamin E in female reproductive aging using multiple experimental methods and multiomics joint analysis.

Methods: The development of follicles was analyzed using Hematoxylin-eosin staining and terminal deoxynucleotidyl transferase deoxyuridine phosphate nick-end labeling staining. The content of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and Estradiol (E2) was determined by Enzyme-linked Immunoassay Kit. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) in the serum using enzyme-linked immunosorbent assay kits. RNA-seq and Untargeted metabolomics analyses were used to investigate the molecular mechanisms of vitamin E during ovarian aging.

Results: The findings revealed that quail reproductive organs rapidly aged at the 11th egg-laying month. Adding vitamin E to the diet significantly improved egg production performance. Vitamin E promoted the development of small yellow follicles by enhancing antioxidant capacity and inhibiting cell apoptosis. Furthermore, RNA-seq and LC-MS/MS identified genes and metabolites that may serve as biomarkers for vitamin E effects.

Conclusion: Dietary supplementation with 250 mg/kg vitamin E enhanced energy metabolism, regulated SOD and MDA levels, reduced apoptosis, stimulated E2 secretion, promoted follicular development, and improved egg production performance.

Background/aims: The feasibility and effectiveness of Tumor Destructive Mechanical Impulse (TMI) treatment of solid tumors for standard clinical application is investigated.

Methods: Different solid tumors in a preparatory animal experiment (VX2 head and neck squamous tumor) and in patients (malignant cutaneous melanoma and prostate carcinoma) are treated by TMI focused shock waves using patient-specific treatment parameters (total energy, energy flux density, shock wave frequency, total number and sequence of shock waves, the optimal placement of the treatment device) determined by multiple parametric simulations.

Results: In animal experiments and in the different treated tumor entities in several patients, the treated tumor or treated metastases regressed, and disguised tumor-associated antigens consistently initiated an immunological abscopal effect achieving that not only the directly treated primary tumor or a treated metastasis regressed but also untreated distant metastases.

Conclusion: TMI treatment could have significant implications for the development of new effective and targeted regimens of cancer therapy.