Cellular Physiology and Biochemistry最新文献

Background/aims: CUL5 acts as the scaffold protein of the E3 ligase complex in ubiquitin-dependent protein degradation pathways. Overexpression of CUL5 inhibits cellular proliferation, whereas inhibition of CUL5 expression induces proliferation and may contribute to clinical disorders including cancer. The effects of CUL5 depend on its post-translational modification by NEDD8 (neddylation), a process that represents a potential therapeutic target. This study explores the structure-function relationship between CUL5 and its neddylation status in vitro.

Methods: CUL5 was mutated at the putative neddylation site Lys (K) 724 to Arg (K724RCUL5) and at three potential neddylation sites K724, K727, and K728 (K724R/K727R/K728RCUL5). Because mutation of the PKA phosphorylation site Ser (S) 730 (S730ACUL5) was previously shown to induce neddylation, a K724R/S730ACUL5 mutant was also generated. Mutant and wild-type constructs were expressed in rat endothelial cells (RAMEC), T47D cancer cells, and COS-1 cells. Cellular proliferation, MAPK phosphorylation, ERα expression, and CUL5 neddylation status were analyzed, including treatment with the neddylation inhibitor MLN4924.

Results: Expression of wild-type CUL5 attenuated cell growth in RAMEC, T47D, and COS-1 cells. In contrast, expression of K724RCUL5 and K724R/S730ACUL5 mutants induced cellular growth, whereas the K724R/K727R/K728RCUL5 mutant had no significant effect on proliferation. In T47D cells, MAPK phosphorylation and estrogen receptor (ERα) expression were directly correlated with the neddylation status of CUL5. Western blot analysis of COS-1 cells treated with MLN4924 demonstrated that CUL5 remained neddylated in all mutant cell lines.

Conclusion: These findings suggest that modification of CUL5 by NEDD8 may occur at multiple lysine residues and that multi-site neddylation may contribute to the diverse regulatory effects of CUL5 on cellular signaling and proliferation.

Background/aims: Nanobiotechnology offers sustainable strategies to enhance plant resistance by activating innate immune responses. This study evaluates the effect of chitosan nanoparticles (CNPs) on transcriptional activation of defense-associated genes in Arabidopsis thaliana.

Methods: CNPs were produced via ionic gelation and sprayed on leaves of 4-week-old plants. Relative expression levels of PR1, PDF1.2, and WRKY70 were measured using qRT-PCR and the 2^-ΔΔCt method with normalization and statistical analysis.

Results: CNP treatment significantly increased PR1, PDF1.2, and WRKY70 expression compared to controls, indicating activation of salicylic acid and jasmonic acid/ethylene signaling pathways. WRKY70 showed the highest induction, suggesting a major regulatory role in coordinating defense signaling.

Conclusion: Chitosan nanoparticles act as effective inducers of transcriptional activation of plant defense markers in Arabidopsis thaliana. Mechanisms including ROS signaling, calcium influx, MAPK cascades, and PTI/ETI responses warrant further investigation.

A strong yet appropriate immune response is essential for the timely control and clearing of respiratory viral infections. However, as physical condition declines with age, so does immune function. Immunosenescence is an age-related immune dysfunction marked by dysregulation of innate immune pathways, thymic involution, decreased T and B cell numbers and function, altered metabolism, and epigenetic changes. Additionally, older individuals often develop inflammaging, a condition characterized by elevated levels of proinflammatory markers. This deterioration of immune function in older adults significantly impairs viral control and increases disease severity. Studies consistently show a progressive rise in hospitalization and mortality rates with age. This review discusses the complex relationship between aging, immunosenescence, and inflammaging during respiratory viral infections, as well as the underlying mechanisms that increase susceptibility, disease severity, and hospitalization among older adults.

Background/aims: To assess the relationship between left ventricular remodeling as evaluated by strain echocardiography and conventional echocardiography before and after cardiac resynchronization therapy (CRT) in patients with severe heart failure.

Methods: A cross-sectional descriptive study with follow-up comparison before and after CRT. The study included 33 heart failure patients indicated for CRT at the Vietnam National Heart Institute, Bach Mai Hospital from late 2015 to December 2021.

Results: Left ventricular end-systolic volume and end-diastolic volume significantly decreased after three months of follow-up (p < 0.001) and showed a strong inverse correlation with QRS recovery (r = 0.65). A positive correlation was observed between changes in left atrial volume index (LAVI) and 4B strain (r = 0.96, p < 0.05). After three months of CRT implantation, positive correlations were observed between isovolumic relaxation time (IVRT) and left atrial volume index (LAVI), and between left atrial volume and E/e' obtained from tissue Doppler imaging (r = 0.53, p < 0.05). No correlation was found between E/e' and total EF, between 2B strain and total EF, or between 2B strain and LAVI.

Conclusion: Strain echocardiography represents an important tool for diagnosing, assessing risk factors, and monitoring heart failure. After three months of CRT implantation, significant improvement in left ventricular volume and ejection fraction was observed, indicating improved treatment effectiveness.

Background/aims: Systemic sclerosis is a rare autoimmune and fibrotic disease, often manifesting in the skin. The aryl hydrocarbon receptor is critical for skin homeostasis; however, its role in fibrosis is not well understood. We investigated the crosstalk between TGFβ, AHR, and Wnt/β-catenin signaling.

Methods: Human dermal fibroblasts and HaCaT keratinocytes, both wild-type and AHR-deficient, were cultured in mono- and co-cultures. Cells were treated with TGFβ and the AHR agonist FICZ. Collagen type I and MMP1 were quantified, Wnt/β-catenin genes analyzed, migration assessed, and proteome profiling performed.

Results: AHR deletion reduced Wnt/β-catenin gene expression and abrogated TGFβ-induced collagen production. Co-cultures showed AHR-dependent regulation of immune-related genes, and scratch closure was also AHR-dependent.

Conclusion: The study demonstrates a context-dependent role of AHR in skin fibrosis and its interaction with TGFβ and Wnt/β-catenin signaling.

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: Infection with Helicobacter pylori is a major cause of chronic gastritis and persistent gastric inflammation. miRNA-155 and inflammatory cytokines play key roles in immune regulation; however, their systemic expression profiles in H. pylori-associated chronic gastritis are not fully characterized.

Methods: Twenty-five patients with endoscopically and histologically confirmed H. pylori-associated chronic gastritis and 25 age-matched healthy controls were included. Peripheral miRNA-155 expression was measured by quantitative real-time PCR, and serum IL-6, IFN-γ, and IL-10 levels were quantified by ELISA.

Results: Patients exhibited significantly elevated peripheral miRNA-155 expression compared with controls (p < 0.001). Serum IL-6, IFN-γ, and IL-10 concentrations were also significantly higher in patients (all p < 0.001). Logistic regression analyses indicated significant associations between biomarker levels and H. pylori infection status.

Conclusion: This exploratory case-control study demonstrates systemic alterations in miRNA-155 expression and inflammatory cytokines in H. pylori-associated chronic gastritis. These findings suggest immune-regulatory changes detectable in peripheral blood, warranting validation in larger independent cohorts.

Sickle cell anemia (SCA) is a progressive, systemic disorder that can lead to multi-organ dysfunction. While it has traditionally been most prevalent in regions where malaria is endemic, recent epidemiological studies have shown an increasing disease prevalence in non-endemic areas, primarily attributed to global human migration patterns. The severity of SCA typically worsens with age. In early childhood, affected individuals may present with renal hyperfiltration, neurocognitive delays, cardiac remodeling, and skeletal fragility. The presence of these early manifestations often predicts the development of chronic complications later in life, including splenic atrophy, neurodegeneration, and impaired cerebral perfusion. Adequate management of SCA begins with universal newborn screening programs, enabling early detection and the initiation of appropriate interventions. Therapeutic advancements, ranging from disease-modifying agents such as hydroxyurea to curative options including gene therapy and stem cell transplantation, have significantly improved clinical outcomes; however, long-term morbidity remains a significant challenge. This review aimed to explore the effect of aging on pathophysiological changes and the onset of organ-specific complications in SCA patients. It highlights the importance of age-tailored monitoring and a multidisciplinary approach to detect early signs of organ damage, prevent irreversible complications, and consequently improve overall quality of life.

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.