Journal of Bioenergetics and Biomembranes最新文献

Colorectal adenocarcinoma (COAD) poses a serious threat to the life of the patient. Notably, Uroplakin 1 A (UPK1A) is a prognostic biomarker for a variety of tumors. However, the role played by UPK1A in the occurrence and development of COAD and its associated molecular mechanisms still lacks a clear and in-depth understanding. The relationship between UPK1A expression and clinicopathological features, as well as patient prognosis, was examined through the use of online databases. Differences in UPK1A expression in COAD tissues and adjacent normal tissues were assessed in clinical samples. The effects of knocking down UPK1A under Escherichia coli (E. coli) co-culture/non-co-culture conditions on COAD cell proliferation, cell invasion, and apoptosis were investigated. In vivo subcutaneous tumor xenograft model, we knocked down the UPK1A gene in a tumor mouse model and assessed tumor growth. The effects of UPK1A and E. coli on glycolysis were investigated by detecting mRNA expression of glucose consumption, lactate production, HIF-1α, and glycolytic enzymes (GLUT1, LDHA, and PDK1). UPK1A was highly expressed in COAD tissues and showed a positive association with unfavorable outcomes in colorectal cancer patients. By knocking down UPK1A, co-culture conditions with E. coli inhibited COAD cell proliferation and invasion, promoted apoptosis, and reduced tumor growth. Knockdown of UPK1A inhibited COAD cell glycolysis by modulating HIF-1α signaling under E. coli co-culture conditions. It is suggested that UPK1A and E. coli synergistically promoted COAD cell proliferation, invasion, and tumor growth and inhibited apoptosis. By regulating HIF-1α signaling, UPK1A and E. coli were able to promote glycolysis in COAD cells. UPK1A and E. coli synergistically interfered with junctional COAD processes.

Metabolic associated fatty liver disease (MAFLD) is a highly prevalent global chronic liver disease. While abnormal expression of RNA-binding proteins (RBPs) has been implicated in MAFLD, their functional roles-particularly in regulating alternative splicing (AS)-remain poorly characterized. This study aimed to investigate the abnormal expression and regulatory mechanism of RBPs in MAFLD. The source data were obtained from the GSE130970 dataset of the Gene Expression Omnibus (GEO) database. Then, we utilized differential expression analysis to acquire the differentially expressed genes (DEGs) between different stages of MAFLD patients and normal patients. Alternative splicing analysis was performed via the ABLas pipeline to explore the alternative splicing events that may enhance and regulate the development of MAFLD. The Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were utilized via KOBAS2.0. Finally, quantitative real-time Polymerase Chain Reaction (RT-qPCR) and Western blotting analysis were performed to confirm the expression of significant RBPs. We observed a significant increase in the number of DEGs as the stages of MAFLD progressed. Furthermore, co-expression analysis suggested that abnormally expressed RBPs such as S100A4, CYCS, and JUN might participate in MAFLD development by potentially influencing the AS of downstream metabolism-related genes such as FAT1, SLCO2B1, and C4BPB. Moreover, we confirmed that the expression of the three RBPs (S100A4, CYCS and JUN) is significantly up-regulated in the liver through validation experiments. The abnormal up-regulated expression of RBPs (S100A4, CYCS and JUN) might contribute to the progression of MAFLD and hence they can be further regarded as potential therapeutic targets for MAFLD.

Liver diseases poses a significant global health burden. This study investigated the hepatoprotective effects of fisetinidin chloride (FC), a natural flavonoid, against carbon tetrachloride (CCl₄)-induced hepatotoxicity in HepaRG cells, a model mimicking human hepatocyte responses. Co-treatment with FC restored cell viability and reduced cellular steatosis, and minimized lactate dehydrogenase leakage, demonstrating membrane stabilization. FC mitigated oxidative stress by reducing mitochondrial reactive oxygen species (ROS) and lipid peroxidation, while enhancing antioxidant defenses through upregulating mitochondrial superoxide dismutase and glutathione. FC preserved mitochondrial function, as evidenced by restored mitochondrial membrane potential (ΔΨm), and modulated apoptosis by upregulating anti-apoptotic BCL2 mRNA and downregulating pro-apoptotic BAX and caspase-3. Flow cytometry analysis confirmed FC's anti-apoptotic effects, reducing apoptotic cell populations. Additionally, FC attenuated CCl₄-induced elevations in aspartate aminotransferase and alanine aminotransferase, markers of hepatocellular injury. Treatment with FC significantly upregulated choline, citric acid, cis-aconitic acid, L-carnitine, L-tryptophan, and gamma-Linolenic acid in CCl₄-induced cells. Conversely, it significantly downregulated glutamate, xanthine, indole acetic acid, succinic acid, hypotaurine, and other metabolites. Pathway enrichment and network analysis of the metabolome demonstrated that FC's protective effects were mediated through the modulation of mitochondrial energy metabolism. Collectively, these findings highlight FC's multifaceted hepatoprotective effects, including attenuation of cellular steatosis, ROS scavenging, mitochondrial stabilization, and apoptosis inhibition. This study underscores FC's potential as a therapeutic candidate worthy of further mechanistic studies, bridging in vitro efficacy with clinical relevance. Further in vivo studies are warranted to validate its pharmacokinetics and translational potential.

Renal ischemia/reperfusion injury (RIRI), a common complication of renal transplantation, partial nephrectomy, and transient hypoperfusion, is a major etiological factor of acute kidney injury (AKI) with limited treatment options. Total flavonoids from Desmodium styracifolium (TFDS), a traditional Chinese medicinal herb used in urinary disorders, have shown promising renoprotective properties. This study aimed to investigate the efficacy of TFDS against RIRI and elucidate its underlying mechanisms, with a particular focus on oxidative stress and ferroptosis. A RIRI model was established in C57BL/6J mice, and the effects of TFDS were evaluated in both in vivo and in vitro hypoxia/reoxygenation (H/R) models. Evaluation of renal function was performed by measuring serum blood urea nitrogen (BUN) and creatinine levels. Histopathological and ultrastructural alterations were examined using hematoxylin-eosin (H&E) staining and transmission electron microscopy (TEM). Oxidative stress and ferroptosis were evaluated by determining glutathione (GSH) levels, malondialdehyde (MDA) content, reactive oxygen species (ROS) levels, and iron accumulation. Potential therapeutic targets and pathways were predicted by network pharmacology and further validated through Western blot (WB) and immunofluorescence analyses. In vivo, TFDS administration markedly improved renal function in RIRI mice, as evidenced by significant reductions in serum BUN and creatinine levels, and attenuated histopathological damage, including tubular epithelial cell loss and mitochondrial structural disruption. TFDS also decreased tissue iron and malondialdehyde (MDA) levels while restoring GSH content, thereby alleviating oxidative stress and ferroptosis. In vitro, TFDS enhanced HK-2 cell viability after hypoxia/reoxygenation injury, reduced intracellular ROS, iron, and MDA accumulation, and preserved mitochondrial morphology. Network pharmacology and molecular docking identified TP53 as a central target, with vicenin-2, schaftoside, and isovitexin exhibiting strong binding affinity to P53. Mechanistically, TFDS downregulated P53 expression and upregulated SLC7A11 and GPX4 both in vivo and in vitro, effects that were abolished by the P53 agonist Kevetrin, confirming the involvement of the P53/SLC7A11/GPX4 axis in TFDS-mediated ferroptosis suppression. TFDS alleviates kidney injury following RIRI by attenuating oxidative stress and suppressing ferroptosis, effects mediated at least in part through modulation of the P53/SLC7A11/GPX4 signaling axis. These findings identify TFDS as a promising therapeutic candidate for ischemic kidney injury and provide mechanistic insight supporting its potential clinical application.

This study aimed to investigate the therapeutic effects of Sini Decoction on a murine model of peripheral arterial disease (PAD) and to explore its potential mechanisms of action related to mitochondrial autophagy and M1 macrophage polarization. A total of 36 specific-pathogen-free Kunming mice were used to establish a PAD model and were randomly assigned into four groups: the experimental group (EG, administered Sini Decoction via gavage), the control group (CG, administered rapamycin via gavage), the model group (MG, administered 0.9% sodium chloride solution via gavage), and the normal group (NG, administered 0.9% sodium chloride solution via gavage). Serum inflammatory cytokines, mitochondrial autophagy-related proteins (LC3bII and p62), M1 macrophage markers (iNOS and COX2), key proteins in the mitochondrial autophagy pathway (PINK1 and Parkin), relative mitochondrial DNA (mtDNA) content, and mitochondrial function indicators [oxygen consumption rate (OCR) and extracellular acidification rate (ECAR)] were measured and analyzed. The serum levels of IL-6, IL-1β, TNF-α, IL-10, and MCP-1 were significantly decreased in both the EG and CG compared to the MG (P < 0.05), with the EG showing considerably greater reductions than the CG (P < 0.05). Compared with the CG, the EG exhibited significantly increased protein and mRNA expression levels of LC3bII, p62, iNOS, and COX2 (P < 0.05), considerably elevated mitochondrial OCR, and considerably reduced ECAR (P < 0.05). Additionally, the relative mtDNA content and the percentage of atherosclerotic lesion area were markedly lower in the EG than in the CG (P < 0.05). Moreover, the expression level of PINK1 and Parkin proteins were significantly increased in both the EG and CG compared to the MG (P < 0.05). Sini Decoction demonstrated superior efficacy in ameliorating PAD compared to the autophagy inducer rapamycin. Its therapeutic effects may be associated with the promotion of mitochondrial autophagy and the induction of M1 macrophage polarization.

Myocardial fibrosis (MF) is a key pathological process driving heart failure, characterized by excessive extracellular matrix (ECM) deposition and impaired cardiac function. Although myocyte-specific enhancer factor 2 A (MEF2A) is implicated in cardiac fibroblast activation, its role in MF remains unclear. We manipulated MEF2A expression in cardiac fibroblasts (CFs) through knockdown and overexpression, and assessed fibrosis markers, migration, and RhoA signaling. Binding of MEF2A to the Snail1 promoter was predicted using JASPAR and validated by chromatin immunoprecipitation (ChIP) and luciferase reporter assays. Rescue experiments with Snail1 overexpression and RhoA inhibition were performed. An angiotensin II (Ang II)-induced MF mouse model was used to evaluate cardiac function by echocardiography and to assess collagen deposition through picrosirius red (PSR) staining. MEF2A was significantly upregulated in Ang II-induced fibrotic hearts and CFs. MEF2A knockdown reduced α-SMA and Col1a1 expression, inhibited CF migration, and suppressed activation of the Snail1/RhoA/α-SMA pathway. ChIP and luciferase assays confirmed the direct binding of MEF2A to the Snail1 promoter. Inhibition of RhoA signaling reversed MEF2A-induced myofibroblast activation and migration. Rescue experiments showed that Snail1 overexpression restored the fibrotic phenotype suppressed by MEF2A knockdown. In vivo, MEF2A knockdown improved left ventricular function, reduced collagen deposition (PSR staining), and lowered heart weight/tibia length ratios. MEF2A promotes myocardial fibrosis by directly activating Snail1 and engages the RhoA/α-SMA pathway. Targeting MEF2A offers a promising therapeutic strategy to attenuate MF and improve heart function.

Clinical evidence points to the Traditional Chinese Medicine Fuzheng Huayu recipe (FZHYR) as an anti-fibrosis drug. Our previous studies have shown that FZHYR regulates macrophage polarization and the expression of NADH dehydrogenase (ubiquinone) 1 alpha subcomplex subunit 2 (NDUFA2) to inhibit pulmonary fibrosis. This study aims to explore the mechanism of FZHYR regulates macrophage polarization and NDUFA2 expression in the treatment of pulmonary fibrosis. NR8383 alveolar macrophages polarizing to M1 or M2 polarization by stimulation with LPS/IFN-γ or IL-14/IL-13 and received FZHYR treatment. Macrophage polarization was verified by detecting the levels of transmembrane protein that specific expression using flow cytometry and levels of inflammatory factors. Oxidative phosphorylation change was reflected by mitochondrial ROS and oxygen consumption rate. The effect of FZHYR on m6A of Ndufa2 mRNA and the involvement of m6A modification enzymes (METTL3 and IGF2BP1) was investigated. FZHYR promoted macrophage M1 polarization and inhibited macrophage M2 polarization. FZHYR inhibited oxidative phosphorylation and NDUFA2 expression in M2 macrophages. Ndufa2 silencing inhibited macrophage M2 polarization and oxidative phosphorylation. M2 macrophage polarization and oxidative phosphorylation induced by Ndufa2 overexpression were reversed by FZHYR. Mechanistically, METTL3 induced Ndufa2 m6A methylation in an IGF2BP1-dependent manner in FZHYR-treated M2 macrophage. Moreover, the inhibition of METTL3 suppressed macrophage M2 polarization and oxidative phosphorylation. FZHYR inhibits M2 macrophage polarization through the inhibition of METTL3-mediated m6A modification and downregulation of NDUFA2 and oxidative phosphorylation.

Heart failure represents the culmination of various cardiovascular diseases, distinguished by a spectrum of complex symptoms. Astragaloside IV (AST-IV) has shown significant cardiac protection in heart failure rats, though the mechanisms are not fully understood. This study aimed to investigate the effects of AST-IV using hypoxia-reoxygenation injury in cardiomyocytes and heart failure in rats to explore the effects of AST-IV. Experimental groups were treated with AST-IV, HIF-2α siRNA, or Y-27,632 (a ROCK inhibitor). Cell proliferation was assessed using CCK-8 and EdU assays, while mitochondrial membrane potential and apoptosis were evaluated using JC-1 fluorescent probes and TUNEL staining, respectively. Additionally, flow cytometry measured reactive oxygen species and apoptosis rates, with protein expressions of HIF-2α, RhoB, and ROCK determined via western blotting. Cardiac troponin I and caspase-3 levels were quantified using ELISA, and myocardial injury was examined through H&E and Masson staining. Results demonstrated that AST-IV notably increased HIF-2α and Rho/ROCK pathway protein expressions, enhancing cell proliferation, reducing apoptosis and ROS levels, but effects were partially reversible by Y-27,632 in vitro. Our findings suggest that AST-IV mitigates hypoxia-induced cardiomyocyte damage by modulating the HIF/Rho/ROCK pathway, indicating its potential as a therapeutic agent for heart failure.

This study investigates the interactions of primary and secondary bile acids (cholic acid (CA), chenodeoxycholic acid (CDCA), ursodeoxycholic acid (UDCA), and lithocholic acid (LCA)) with isolated rat liver mitochondria, focusing on their uncoupling activity, detergent effects, and antioxidant properties. Using a recently developed methodological approach based on quantifying the effective distribution coefficient ([Formula: see text]), we precisely assessed the partitioning of bile acids between the mitochondrial and aqueous phases. Our results demonstrate that the uncoupling potency rank order was LCA > CDCA > CA, which strongly correlated with their lipophilicity. In contrast, UDCA, which possesses hydroxyl groups on the hydrophobic β-surface, exhibited significantly lower uncoupling activity. At concentrations inducing mild uncoupling (stimulating state 4 respiration by 70-75%), all bile acids significantly reduced the ADP/O ratio and respiratory control ratio without inhibiting the electron transport chain, confirming their protonophoric mechanism. Furthermore, we quantitatively showed that bile acids, in contrast to palmitic acid, exert a mild detergent effect, as evidenced by a increase in NADH-stimulated respiration, with UDCA and CA having the most pronounced effect. Crucially, at these uncoupling concentrations, all bile acids consistently suppressed mitochondrial H₂O₂ generation by 30-40%, revealing their antioxidant potential. These findings provide quantitative insights into the structure-dependent dual roles of bile acids in modulating mitochondrial function.

According to reports, Rab3A plays a kay role in various diseases. The regulatory role of Rab3A in hair cell damage and age-related hearing loss has not been explored. HEI-OC-1 cells were treated with hydrogen peroxide (H₂O₂) to construct a damage model. The cell viability and apoptosis were detected by CCK-8 assay and flow cytometry. Immunofluorescence and flow cytometry were used to measure mitochondrial membrane potential. The contents of oxidative stress-related indicators and mitochondrial function-related indicators were detected by kits. Dual luciferase assay was used to determine the regulatory relationship between Rab3A and ITGA3. The results showed that H₂O₂ treatment reduced the level of Rab3A in HEI-OC-1 cells in a time-dependent manner. Rab3A increased the cell viability of H₂O₂-induced inner ear cells and inhibited cell apoptosis. Additionally, Rab3A inhibited H₂O₂-induced oxidative stress and alleviated mitochondrial dysfunction in HEI-OC-1 cells. Rab3A also directly targeted and positively regulated ITGA3 expression. Further studies found that silencing of ITGA3 reversed the inhibitory effects of Rab3A on inner ear cell damage and mitochondrial dysfunction. In conclusion, Rab3A regulates H₂O₂-induced inner ear hair cell damage and mitochondrial dysfunction by stabilizing the expression of ITGA3.