Molecular and Cellular Biochemistry最新文献

Venous thromboembolism (VTE) is a prevalent and potentially fatal condition, typically initiating deep vein thrombosis (DVT). Subsequent embolization can lead to pulmonary embolism (PE), a significant threat to cardiovascular function. Approximately 5% of PE cases result in sudden cardiac arrest due to acute right ventricular failure, arrhythmias, and circulatory shock. Given the considerable influence of PE on right ventricular function, this study aimed to investigate alterations in gene expression within the right ventricle associated with PE in a porcine model. The study was performed on 12 male pigs, divided into two groups of six animals each. In the experimental group, a thrombus was induced in the right femoral vein of each animal and subsequently released to induce pulmonary embolism. The remaining six animals served as a control group. Right ventricular myocardial samples were collected for subsequent transcriptomic and immunohistochemical analyses. In the experimental group, tissue samples were taken 24 h post-thrombus release to characterize changes typical of acute PE. Multistep bioinformatics identified 1992 significantly differentiating molecules: 1436 differentially expressed genes (DEGs), 308 differentially expressed lncRNAs (DELs), and 248 other RNA species. Among these, 417 DEGs, 101 DELs, and 58 other RNAs were downregulated, while 1019 DEGs, 207 DELs, and 190 other RNAs were upregulated. Gene Ontology (GO) analysis assigned molecules to 259 processes (184 biological, 37 cellular, 33 molecular), indicating active inflammatory and immune signaling in cardiac tissue, suggesting roles in ventricular remodeling and stress response. KEGG pathway analysis highlighted altered Oxytocin signaling and Complement and coagulation cascades in PE hearts. Splicing analysis revealed 55,950 alternative splicing events in Sus scrofa right ventricles (PE vs. CTR). Additionally, 3142 single nucleotide variants (SNVs) were identified. This study presents the first comprehensive analysis of right ventricular gene expression profiles in response to PE. The findings indicate that alterations in the oxytocin signaling pathway, the complement and coagulation cascades, and disturbed inflammatory and immune signaling are critically involved in the pathogenesis of right ventricular dysfunction during PE.

Tumor metastasis is the primary cause of high mortality rates among cancer patients. Invasive tumor cells disseminate through the bloodstream as circulating tumor cells (CTCs) and subsequently colonize secondary organs. In addition to genetic mutations that govern biological processes, epigenetic alterations also regulate tumor cell gene expression and chromosome stability. Dysregulation of these patterns is pivotal to carcinogenesis, tumor progression, and metastasis. This narrative review initiates with a comprehensive overview of the biology of CTCs and the processes that contribute to epithelial-mesenchymal transition (EMT), immune surveillance, and colonization. Subsequently, the impact of epigenetic modifications in CTCs on these metastatic processes during CTC migration, survival, and dissemination will be examined. In addition, the paper presents the implications of targeting epigenetics in CTCs for cancer patients and introduces CTCs as new biomarkers for early detection, prognosis, and targeted therapies to improve patient outcomes.

Pharmacological interventions to inhibit the progression of aortic aneurysm (AA) have not yet been established. We previously reported that mesenchymal stem cells (MSCs) provide a potential foundation for less invasive treatment of AA. In this study, we investigated the secretory proteins from MSC supernatants to clarify the therapeutic effects of MSCs. Furthermore, we treated thoracoabdominal aortic aneurysm (TAAA) mice with two anti-inflammatory proteins from among these secretory proteins to confirm their therapeutic effects. Protein profiles of MSC-secreted factors were analyzed using protein microarrays, and two anti-inflammatory proteins, namely progranulin (PGRN) and secretory leukocyte protease inhibitor (SLPI), were identified. Apolipoprotein E-deficient mice were continuously infused with angiotensin II via an osmotic pump for 4 weeks to induce TAAA formation, and then recombinant rPGRN and/or rSLPI were administered intraperitoneally. Mice were sacrificed at 8 weeks, and aortas were analyzed for protein expression and also stained with Elastica van Gieson and immunofluorescence to detect inflammatory cells. Intraperitoneal administration of rSLPI inhibited TAAA growth more than rPGRN alone or the combination of rPGRN and rSLPI, by inducing the following effects: downregulation of inflammatory cytokines and chemokines, specifically IL-1β, IL-6, TNF-α, and MCP-1; reduced NO production; decreased phosphorylated NF-κB levels; and decreased elastin destruction and infiltration of inflammatory cells. We identified anti-inflammatory proteins, including PGRN and SLPI, in the MSC supernatants and showed that the administration of rSLPI inhibited TAAA progression in mice. These promising preliminary data present a new approach for the treatment of less invasive TAAA.

Diabetic retinopathy (DR) is an irreversible microvascular complication in individuals with diabetes. Kaempferol, a flavonoid with anti-inflammatory, antioxidant, and hypoglycemic activities, has exhibited therapeutic potential in previous investigations for treating DR. However, its accurate molecular mechanisms remain elusive. This study aimed to elucidate similarity underlying the progression of DR from early to late stages, along with exploring the key targets of kaempferol for DR therapy. Combined with weighted gene co-expression network analysis (WGCNA) and single-cell RNA sequencing (scRNA-seq) analysis, we elucidated hub regulatory genes and cell subpopulations. Molecular docking was conducted to analyze molecular interactions. Evans Blue (EB) leakage assay, Hematoxylin & Eosin (H&E) and Periodic Acid-Schiff (PAS) staining was utilized to assess retinal structural and vascular damage. Additionally, TUNEL staining was applied to evaluate retinal apoptosis. Comprehensive analyses, including enzyme-linked immunosorbent assays (ELISA), immunofluorescence, Western blotting, and real-time PCR were employed to monitor cytokine levels and protein expression. Our findings preliminarily unveiled that kaempferol could modulate the P21/Thioredoxin pathway, and exerted protective effects on DR by regulating metabolism disorder and cellular dysregulation. Moreover, a novel mechanistic connection was established between fibroblasts activity and DR fibrosis progression, underscoring the pivotal role of the VCAM signaling pathway in vascular cell regulation and its contribution to disease pathogenesis. This study provides new perspectives on the therapeutic potential of kaempferol in DR, particularly regulating vascular injury and cellular senescence via the P21/Thioredoxin axis, which expand the horizon of natural compounds in addressing the vision-threatening complications associated with diabetes.

Gestational diabetes mellitus (GDM) is a prevalent metabolic disturbance in pregnancy. This study analyzed the mechanism of maternal GDM inducing myocardial injury in male offspring through growth differentiation factor-15 (GDF-15). Pregnant rats were randomly assigned to the GDM-mother (streptozotocin [STZ] induction) and the Control-mother (normal saline injection) groups. Here, 32 male offspring from the Control-mother group and 92 from the GDM-mother group were used for experiments. The myocardial ischemia model was established by left anterior descending (LAD) coronary artery ligation in 6-week-old male offspring. Male offspring in the GDM-mother group were treated with sh-Gdf15, pyrroloquinoline quinone, or rotenone. Cardiac function, oxidative stress-associated indicators, myocardial infarct size and necrosis, inflammatory infiltration, cardiomyocyte apoptosis, mitochondrial damage, and Gdf15 mRNA and protein expression were examined using echocardiography, kits, TTC/H&E/TUNEL staining, flow cytometry, RT-qPCR, and western blot. GDM maternal rats had elevated blood glucose and a reduced body weight, representing successful modeling. Prenatal STZ exposure did not affect blood glucose but decreased the body weight in male offspring. The baseline cardiac function was not affected by prenatal STZ exposure, whereas LAD ligation-induced ischemia caused severe cardiac dysfunction in GDM male offspring versus controls. GDF-15 was upregulated in GDM rat male offspring, and its knockdown alleviated myocardial injury. Adult male offspring of GDM rats exhibited pronounced mitochondrial damage, and mitochondrial homeostasis restoration improved ischemia-caused cardiac dysfunction. Suppressing mitochondrial function partly abrogated cardioprotective effects of Gdf15 knockdown. Maternal GDM promoted myocardial injury in male offspring by upregulating GDF-15 to aggravate mitochondrial damage.

Rhynchophylline (Rhy), a bioactive alkaloid extracted from Uncaria species (Uncaria rhynchophylla and Uncaria tomentosa), has demonstrated therapeutic potential in neurodegenerative and cardiovascular diseases due to its diverse pharmacological effects. However, its role in the development or treatment of atherosclerosis has not yet been studied. In this study, we evaluated the anti-atherosclerotic effects of Rhy using both in vivo and in vitro models. In high-fat diet-fed New Zealand White rabbits, Rhy treatment significantly reduced aortic plaque progression, improved vascular histology, and decreased serum cholesterol levels. In THP-1 macrophages, Rhy inhibited ox-LDL uptake and subsequent foam cell formation by lowering scavenger receptor expression. In endothelial cells, it decreased the expression of adhesion molecules, thereby reducing monocyte adhesion and transendothelial migration. Mechanistically, Rhy suppressed the activation of MAPK and NF-κB signaling pathways, contributing to its anti-inflammatory and antioxidant effects. Overall, these results demonstrate that Rhy offers multi-targeted protective effects against atherosclerosis and could be a promising candidate for its prevention and treatment.

Cardiac fibrosis results from cardiovascular diseases (CVDs) and is characterized by excessive extracellular matrix (ECM) accumulation, particularly collagen, leading to cardiac dysfunction. Despite the availability of treatments for CVDs, no targeted therapies are available to prevent disease progression to irreversible stages. Further research is needed to explore the pathways and signaling molecules involved in this progression. The renin-angiotensin system (RAS) plays a significant role, with pharmacological agents targeting its harmful axis, i.e., Angiotensin-converting enzyme (ACE)/Angiotensin II (Ang II)/Angiotensin 1 receptor (AT1R) axis, while an antagonistic axis, ACE2/Angiotensin 1-7 (Ang 1-7)/mitochondrial assembly receptor (MasR) axis offers cardioprotective effects. Reactive oxygen species (ROS) also contribute to CVDs, with NADPH oxidases (NOXes) being key inducers of ROS. NOX1, NOX2, NOX4, and NOX5 are upregulated in pathological conditions, exacerbating the disease. This review focuses on the mechanisms by which the ACE/Ang II/AT1R and ACE2/Ang 1-7/MasR axes regulate NOX activity, aiming to enhance our understanding of future targeted therapies.

Atherosclerosis (AS) is a major cardiovascular disorder, with challenges in early diagnosis and a lack of individualized treatment that require urgent attention. This study employed bioinformatics approaches to identify critical genetic markers linked to AS pathogenesis and explored their underlying molecular mechanisms to facilitate advancements in diagnostic accuracy and therapeutic interventions. We successfully identified genes exhibiting significant differential expression in AS, i.e., oxidative stress and glycolysis-related differentially expressed genes (OSGRDEGs). Through weighted gene co-expression network analysis, three modules (MEturquoise, MEred, and MEgreen) significantly associated with AS were screened, and 72 module genes were found to be identical to OSGRDEGs. A protein-protein interaction network was designed through comprehensive integration of data from the STRING database, followed by visualization and topological analysis employing Cytoscape software. Candidate genes were further evaluated using five distinct algorithms within the CytoHubba plugin, resulting in 12 high-confidence hub genes associated with AS pathogenesis. The 12 hub genes screened by machine algorithm were further screened by modeling to obtain 7 key genes. Finally, statistical analysis revealed marked variations in the infiltration levels of eight immune cell populations across the comparative groups. Monocytes and M0 macrophages showed significant negative correlations in subtypes A and B. Notably, APOE and CXCL1 demonstrated strong positive associations with M0 macrophages and monocytes, respectively, as evidenced by our correlation analysis. This study highlights the use of a bioinformatics approach to identify molecular markers of AS, with future work focused on validating their potential clinical applications.

Immune checkpoint inhibitors (ICIs), especially anti-PD-1 immunotherapy, offer a new treatment option for tumor patients. However, its efficacy is limited in the most of patients with immunologically "cold" tumors. An important histone demethylase, histone lysine-specific demethylase 1 (LSD1/KDM1A), plays a significant role in T cell regulation. Combining LSD1 inhibitors with anti-PD-1 mAb has shown improved anti-tumor effects in various solid tumors. Specifically, in gastric cancer (GC), LSD1 knockdown boosts T cell-mediated anti-tumor immunity. Nevertheless, currently, this effect is only related to PD-L1 in exosomes. Therefore, further research on the molecular mechanisms of LSD1 in regulating T cells in GC is needed. Using TIMER 2.0 and GEPIA 2 databases, we analyzed LSD1 expression in GC and its gene correlations. Lentiviral transfection was utilized to construct a control cell line (shControl) and an LSD1 knockdown cell line (shLSD1). The mRNA and protein levels of LSD1 and immune-related cytokines were measured through real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and Western blotting. We examined the role of LSD1 knockdown in regulating T cell anti-tumor immunity via transcriptome sequencing (RNA-seq). Mouse subcutaneous graft tumor models and in vitro conditioned culture models were established, and the altered functional phenotypes of T cells in mice and in vitro were assessed by RT-qPCR and flow cytometry. Genetic inhibition of LSD1 in GC cells increased T cell proliferation, CD8⁺ activation, and chemotaxis in vitro. Pharmacological inhibition of LSD1 curbed tumor growth in vivo. Remarkably, the combination LSD1 inhibitors with PD-1/PD-L1 blockers led to greater efficacy. At the molecular level, LSD1 knockdown induced the transcription of tumor necrosis factor ligand superfamily member 14 (TNFSF14). As a result, T cell-mediated anti-tumor immunity was improved. Inhibiting LSD1 in GC upregulates TNFSF14 expression, which in turn promotes T cell proliferation, CD8⁺ activation, and chemotaxis. This enhancement of T cell-mediated anti-tumor immunity is further amplified when LSD1 inhibitors are used alongside PD-(L)1 blockers, facilitating the activation of CD8⁺ T cells in the spleen and improving leukocyte infiltration in the tumor.