Molecular and Cellular Biochemistry最新文献

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.

MiR-150-5p is a microRNA that plays an important role in the heart diseases. However, its specific role and molecular mechanism in heart failure (HF) remain unclear. In this study, we found that miR-150-5p was downregulated in patients with HF, while the expression of MMP14 was elevated. In vitro experiments have shown that miR-150-5p directly targets MMP14 and inhibits its expression in human aortic smooth muscle cells (HASMCs). Functionally, miR-150-5p promotes the proliferation, migration and apoptosis of HASMC, which is a key process in the progression of HF. In the HF mouse model induced by transverse aortic contraction (TAC), overexpression of miR-150-5p can improve cardiac function, reduce hypertrophy and fibrosis, while its inhibition aggravates these effects. These findings determined that miR-150-5p is a protective regulator in HF, functioning by inhibiting MMP14, and indicated its potential as a therapeutic target for HF treatment.

Sorafenib targets various tyrosine kinase receptors, inhibiting cell growth and proliferation, angiogenesis and metastasis in tumour cells. It is used to treat certain types of cancers including renal, thyroid and liver (hepatocellular carcinoma) cancers. Although Sorafenib is approved for advanced hepatocellular carcinoma, it only extends patient´s lives by a few months, highlighting the urgent need to better understand how it works and to develop more effective treatments. Sorafenib specifically inhibits translation initiation in hepatocellular carcinoma cells. Herein, we revealed that this inhibition results, at least, from the activation of PERK, triggering a stress response that leads to eIF2α phosphorylation, the inhibition of MNK1a-signalling-dependent eIF4E phosphorylation, and the aberrant assembly of the canonical eIF4F complex. Sorafenib also inhibits the ERK1/2 MAPK signalling in HepG2 cells. However, the mTORC1 pathway does appear to play a pivotal role in Sorafenib-dependent translation inhibition, as revealed by the phosphorylation levels of RPS6 and 4EBP1 proteins and the effects on translation of gene silencing 4EBP1/2 in Sorafenib-treated cells. Translation inhibition correlates with reduced production of cancer-promoting proteins like Cyclin D1 and c-Myc. Overexpression of the phosphomimetic eIF4E-S209D variant, which constitutively activates eIF4E, shows that inhibition of eIF4E phosphorylation directly causes Cyclin D1 down-regulation and cell-cycle delay in Sorafenib-treated cells. Taken together, our results confirm that Sorafenib induces translation reprogramming, whose understanding is crucial for improving its efficacy as a cancer therapy.

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.