Journal of Bioenergetics and Biomembranes最新文献

As an important transcription factor, activin receptor-like kinase 5 (ALK5) plays a crucial role in the development of various diseases. However, there have been no reports on whether ALK5 is involved in the pathogenesis of asthma, and further exploration is needed. An in vitro asthma model was constructed using house dust mite (HDM). Quantitative real-time polymerase chain reaction and western blot were used to detect the expression of ALK5. Enzyme-linked immunosorbent assays were used to measure the levels of inflammatory factors. Oxidative stress-related factors and glycolysis-related indicators were analysed using commercial kits. Co-immunoprecipitation was used to assess the binding activity of ALK5 and Krüppel-like factor 4 (KLF4). Finaly, an HDM-induced asthmatic mouse model was established. H&E and PAS staining were used to evaluate the pathological status of mouse lungs, immunohistochemistry (IHC) was used to detect the expression of ALK5 and KLF4 in mouse lung tissue, and Masson staining was used to detect collagen deposition. The results showed that ALK5 was up-regulated in HDM-induced BEAS-2B cells. Silencing ALK5 suppressed inflammation and apoptosis in BEAS-2B cells. Furthermore, knockdown of ALK5 inhibited oxidative stress and promoted glycolysis in BEAS-2B cells. ALK5 specifically bound to KLF4 and promoted its protein degradation. Mechanistically, KLF4-mediated glycolysis was involved in the regulation of ALK5 in BEAS-2B cells. In vivo, ALK5 knockdown attenuated airway inflammation, reduced inflammatory cell infiltration, decreased collagen deposition, and improved lung histopathological damage. In conclusion, inhibiting ALK5 suppresses inflammation, apoptosis, and oxidative stress by regulating KLF4-mediated glycolysis in BEAS-2B cells, thus inhibiting the further development of asthma. Therefore, ALK5 may be a potential target for the clinical treatment of asthma.

Ischemia/reperfusion-induced myocardial dysfunction remains a clinical problem and is associated to poor outcomes in patients with cardiovascular disorders, such as myocardial infarction. Shionone is a triterpenoid extracted from the herbal medicine Radix Asteris which has health benefits. This study aimed to explore the roles and functional mechanism of Shionone in regulating myocardial ischemia/reperfusion injury. The network pharmacology was performed to analyze the potential pathway interacted with Shionone in myocardial ischemia/reperfusion injury. The oxygen-glucose deprivation and reperfusion (OGD/R)-treated H9c2 cardiomyocytes and ischemia/reperfusion-induced murine models were regarded as in vitro and in vivo models. Lactate dehydrogenase (LDH), reactive oxygen species (ROS), glutathione (GSH) and iron levels were analyzed using specific kits. Related protein levels were detected by immunofluorescence staining and western blotting assays. Heart infarct in mice was investigated via TTC staining. Network pharmacology predicted LCN2 and PI3K/Akt signaling might be required by Shionone to involve in myocardial ischemia/reperfusion injury. Shionone mitigated OGD/R-induced ferroptosis through decreasing LDH release, ROS generation, iron and ACSL4 levels, and enhancing GSH, SLC7A11 an GPX4 levels in cardiomyocytes. Shionone attenuated OGD/R-induced endoplasmic reticulum stress through reducing CHOP, GRP-78, and phosphorylation levels of PERK and eIF2α. Endoplasmic reticulum stress inducer reversed the effects of Shionone on cardiomyocyte ferroptosis. Shionone decreased LCN2 expression and enhanced activation of PI3K/Akt signaling. Overexpressed LCN2 reversed the effects of Shionone on cardiomyocyte ferroptosis and endoplasmic reticulum stress in OGD/R-treated cardiomyocytes, and this function was mitigated via PI3K/Akt signaling activation. Shionone mitigated ischemia/reperfusion damage in murine heart by reducing heart infarct. Shionone attenuated endoplasmic reticulum stress-associated ferroptosis in cardiomyocytes through decreasing LCN2 and activating PI3K/Akt signaling, offering a basis for understanding the potentially cardioprotective potential of Shionone post ischemia/reperfusion injury.

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.