Relationship between β-arrestin1-GAPDH interaction and the DADLE-mediated protection of brain microvascular endothelial cells from hypoxia-ischemic/reperfusion injury.

Curcumol induces autophagy-dependent hepatic stellate cell death through methionine metabolism disruption.

Lead (Pb), a persistent and highly toxic environmental contaminant, induces severe multi-organ toxicity, with the liver as a primary target. Naringenin (Nar), a natural flavonoid with established antioxidant properties, shows potential in mitigating chemical-induced liver injury; however, its protective mechanism against Pb hepatotoxicity remains unclear. This study investigated the hepatoprotective effects and underlying mechanisms of Nar in Pb-exposed rats. Thirty-six five-week-old male Sprague-Dawley rats were randomly assigned to six groups (n = 6): control, three Pb-treated groups (15, 30, and 60mg/kg), co-treatment group (60mg/kg Pb + 50mg/kg Nar), and Nar-alone group (50mg/kg). After 8 weeks, Pb exposure induced significant liver injury, evidenced by histopathological lesions (e.g., central venous congestion and inflammation), elevated serum levels of ALT, AST, and LDH, and oxidative stress (increased MDA; decreased CAT, T-SOD, and GSH). At the molecular level, Pb exposure suppressed the Nrf2/NQO1 pathway, upregulated Keap1, and impaired autophagic flux, as indicated by a decreased LC3-II/LC3-I ratio, accumulated p62, and a disrupted PINK1/Parkin pathway. Corroborating the impaired flux, transmission electron microscopy (TEM) revealed an accumulation of autophagic vacuoles, indicative of blocked autophagic degradation. Crucially, Nar co-treatment effectively mitigated these Pb-induced disturbances by activating the Nrf2/NQO1 pathway to counteract oxidative stress and restored autophagic flux, which was supported by the reversal of p62 accumulation and normalization of the LC3-II/LC3-I ratio. In conclusion, Nar protects against Pb-induced hepatotoxicity by concurrently alleviating oxidative stress and restoring autophagic function, supporting its potential as a therapeutic agent.

A multimodal ROS logic-gated therapeutic platform disrupts the vicious cycle of senescence to promote aged bone defect repair.

ABSTRACT Lung cancer is a common cause of cancer-related mortality worldwide. Luteolin, a naturally occurring flavonoid compound, was reported to exert anticancer effects through various mechanisms: A549 cell non-small cell lung cancer (NSCLC) serves as a model system. The objective of this study was to investigate the effects of luteolin on apoptosis and autophagy using A549 cells. A549 cells were treated with varying concentrations of luteolin for 12, 24, or 48 hr, and cell viability was assessed using the CCK-8 assay to determine optimal concentrations and exposure times. After 24 hr incubation with 10, 40, or 160 μM luteolin, the influence of apoptosis and autophagy was determined using Hoechst 33,258 and MDC staining, respectively. Protein expression levels of Bcl-2, Bax, caspase-3, PI3K, AKT, and mTOR were measured by Western blot. Results demonstrated that luteolin significantly inhibited A549 cell proliferation in a concentration- and time-dependent manner. At all concentrations, luteolin significantly (1) promoted apoptosis and autophagy, (2) increased expression levels of Bax and cleaved caspase-3, (3) suppressed Bcl-2 expression, (4) enhanced autophagic vacuole formation, and (5) inhibited phosphorylation of PI3K, AKT, and mTOR pathways. Data suggest that luteolin inhibits proliferation of A549 cells by regulating apoptosis and autophagy.

Breast cancer (BRCA) remains a major health burden. Ferritinophagy, a selective form of autophagy regulating iron homeostasis, has been linked to tumor progression. This study aims to explore ferritinophagy-associated biomarkers and develop a prognostic model in BRCA through integrative multi-omics analysis. 30 differentially expressed ferritinophagy-related differentially expressed genes (FeRDEGs) were detected in BRCA, followed by functional enrichment analyses to investigate their biological significance. The prognostic risk model was formulated by integrating Cox and LASSO regression analyses. Patients were stratified based on calculated risk scores, and model performance was validated in an independent cohort. Additionally, we assessed mutation profiles, drug-gene interactions, and immune infiltration characteristics. Functional annotation and clustering revealed links to cellular stress responses, autophagy regulation, and metabolic remodeling. Through univariate Cox and LASSO regression analyses, ATG5, JUN, and TFRC were identified as significantly associated with patient survival. A multigene prognostic model was constructed based on their expression, which facilitated risk stratification of patients with statistically significant differences in survival outcomes. The two risk groups also exhibited different mutational landscapes, druggable gene profiles, and drug sensitivity, suggesting implications for personalized therapy. Immune infiltration analysis offered additional contextual support for the findings. The multi-gene prognostic model based on ATG5, JUN, and TFRC demonstrated stable risk stratification and potential clinical utility in breast cancer. Future validation with larger, multi-center cohorts and additional multi-omics features could further enhance its predictive performance. We explored the biological relevance of FeRDEGs and established a prognostic model based on ATG5, JUN, and TFRC. The model was validated across multiple dimensions, supporting its potential value in risk stratification and clinical application. These findings might contribute to offering a foundation for exploring therapeutic strategies targeting iron metabolism and autophagy-related pathways.

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal lung disease for which effective therapies are lacking. While cellular senescence and autophagy have been implicated in IPF pathogenesis, the intersecting mechanisms that drive fibrosis progression remain unclear. Our patient-derived transcriptomic analysis revealed a strong association between enhanced extracellular matrix remodeling, cellular senescence, and autophagic dysfunction in IPF. Among the potential driver genes associated with cellular senescence, the elevated expression of p16INK4A (p16) is correlated with impaired autophagic flux in naturally aging mice, bleomycin (BLM)- and radiation-induced pulmonary fibrosis (PF) mouse models, and lung tissues from IPF patients. Genetic depletion of p16 reduced PF and preserved autophagic flux in response to fibrotic challenge, independent of p21Cip1. Applying this knowledge, we screened antifibrotic compounds from extracts of the medicinal fruit of Melia azedarach and identified toosendanin. This compound suppressed p16 promoter activity, effectively restored autophagic flux, and attenuated BLM-induced PF in vivo. We further showed that the p16 promoter inhibitor abyssinone II alleviated both autophagy dysfunction and fibrosis progression, whereas the promoter activator neorautenol exacerbated both phenotypes, demonstrating that p16 acts as a key regulatory node in the regulation of the crosstalk between senescence and autophagy dysfunction. Collectively, our results identify p16 as a mechanistically defined regulator of fibrosis progression and suggest that modulating this axis may offer new opportunities for therapeutic intervention in IPF.

Dysregulation of podocyte autophagy, triggering serious impairment of podocyte integrity, is a possible determinant of IgAN. To decipher the significant roles of Beclin-1 and LC3B in IgAN in regulating cellular autophagy, we investigated the role of PI3K/AKT signaling in regulating podocyte autophagy of IgAN. To carry out this study, we obtained tissue samples from 80 patients with primary IgAN (IgAN group) and 20 patients with kidney stones (control group). The IgAN group was further classified according to the Haas pathological grading system. The 100 paraffin-embedded tissue samples underwent hematoxylin-eosin staining, immunofluorescence, special staining techniques, immunohistochemical analysis (AKT, FoxO1, Beclin-1, LC3B, nephrin and podocin), and transmission electron microscopy. The number of autophagosomes was generally lower in the IgAN group than in the control group, however, in the IgAN group, the number of autophagosomes showed an increasing trend, with higher Haas grade. Beclin-1 and LC3B expression levels were highly correlated, and they were positively correlated with Haas grading, with LC3B showing a "U-shaped" trend: initial low expression in early grades (I-II), progressive increase to peak at grade III, followed by a potential decline in advanced grades (IV-V). Meanwhile, AKT expression was markedly reduced in the early stages (grades I-II) but gradually increased in the late stages (grades III-V). The number of autophagosomes demonstrated a strong positive correlation with Beclin-1 and LC3B expression. Autophagy is a key player in the occurrence and progression of IgAN. Dysregulation of the autophagy pathway represents a core mechanism in the pathogenesis of IgAN, a finding supported by the strong positive correlation between Haas grading and the number of autophagosomes, as well as Beclin-1/LC3B expression.

Fabry disease (FD) is a lysosomal storage disorder caused by a deficiency of α-galactosidase A. Although the accumulation of glycosphingolipids occurs very early, clinical manifestations are often delayed. Little is known about the cellular changes that occur in the presymptomatic phase and may progressively cause damage. To this aim, we evaluated inflammatory markers, cellular stress, and autophagy in presymptomatic pediatric patients with FD identified through newborn screening. We conducted a cross-sectional study of 12 male presymptomatic FD patients with a mean age of 3.11years. We measured plasma cytokine levels (IL1β, TNFα, and IL6) by ELISA. In six patients we also analyzed cellular stress (p-p38) and autophagy markers (LC3, Beclin, Atg3, Atg5, Atg7, Atg12, and Atg16L) by Western blot in peripheral blood mononuclear cells. We observed elevated levels of IL1β and TNFα in 10/12 and 11/12 patients, respectively. P-p38 activation increased by an average of 1.31-fold (SD 1) compared to controls. Autophagy markers showed significant reductions, with LC3-II levels decreasing by an average of 53% (SD 54%). Upstream autophagy mediators (Beclin, Atg3, Atg5, Atg7, Atg12, and Atg16L) were also reduced. This study is the first to demonstrate the presence of inflammation and autophagy inhibition in pediatric presymptomatic FD patients. These findings provide new insights into the early pathogenesis of FD and suggest potential biomarkers for disease progression.

Nuclear receptors in age-related diseases: From mechanisms to drug discovery.

DangGui NianTong Decoction attenuates intervertebral disc degeneration by mitigating nucleus pulposus cell senescence and apoptosis through autophagy induction.

CEACAM1 regulates autophagy in oral squamous cell carcinoma through the PPAR signaling pathway.

Functional characterization of EcPSMB10: Suppression of RGNNV replication via inhibiting autophagy in vitro.

Building the autophagosome: Molecular logic and membrane dynamics of autophagy.

SUMMARYNearly all cellular processes are pH dependent. The acidic pH inside the lysosome (vacuole in yeast) is essential for cellular content degradation, signaling, and autophagy. Defects in lysosome/vacuole acidification are a conserved hallmark of aging and age-related diseases. Traditionally, the lysosome/vacuole is thought to import free protons (H+) from the surrounding neutral cytosol. Here, we uncovered a conserved lysosome/vacuole acidification mechanism from yeast to human involving lysosomal/vacuolar uptake of H+ pumped out by mitochondrial electron transport chain through mitochondria-lysosomes/vacuoles membrane contacts. Aging/senescence-associated disruption of mitochondria-lysosome/vacuole contacts causes lysosomal/vacuolar de-acidification, which can be reversed by either expressing an engineered linker to connect these two organelles or through an asymmetry-dependent rejuvenation process in daughter cells. Preserving lysosomal acidification in senescent human cells prevents the induction of major senescence-associated secretory phenotype factors and restores autophagic flux. These findings reshape our current understanding of the mechanisms underlying lysosomal/vacuolar (de-)acidification in both young and aged/senescent cells.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies, with rising incidence and mortality rates, in which autophagy plays a pivotal role in promoting tumor survival. Although SEC61G has been reported to be associated with poor outcomes in patients with PDAC, its underlying mechanism remains largely unclear. In the present study, we found that SEC61G was overexpressed in PDAC and correlated with an unfavorable patient prognosis. Further experiments demonstrated the pro-proliferation efficacy of SEC61G in PDAC progression both in vitro and in vivo. Mechanistically, SEC61G enhanced cellular autophagy in PDAC in a Ca2+ leakage-dependent manner, which could be reversed by thapsigargin treatment. Interestingly, we unveiled that the elevated intracellular Ca²⁺ concentration mediated by SEC61G could activate CREB, phosphorylated CREB bound to the SEC61G promoter and consequently enhanced SEC61G expression. This positive feedback loop may account for the persistent activation of autophagy in PDAC. Our findings highlight the potential of SEC61G as a prognostic biomarker and a therapeutic target candidate for combating PDAC.

ObjectivesTo characterise the myocardial transcriptomic landscape of patients with systemic sclerosis (SSc) with primary heart involvement (pHI) and identify molecular pathways underlying its pathogenesis.DesignA single-centre study exploring the molecular pathogenesis of SSc-pHI through transcriptomic analysis of endomyocardial biopsy specimens.SettingBasic research.ParticipantThis study enrolled seven patients with SSc-pHI and eight patients with dilated cardiomyopathy (DCM) who had undergone endomyocardial biopsy for clinical practice purposes.InterventionsNo intervention.Main outcome measuresNot applicable.MethodsEndomyocardial biopsy specimens from patients with SSc-pHI and those with DCM underwent whole RNA sequencing. To enable indirect comparison with non-failing (NF) myocardium, public RNA sequencing data of NF and DCM samples were integrated using DCM as a shared reference. Differential gene expression, pathway enrichment (Ingenuity Pathway Analysis and Gene Set Enrichment Analysis) and immune and stromal cell deconvolution were performed. Histopathological evaluation included LC3 immunostaining and transmission electron microscopy (TEM).ResultsA total of 700 genes were differentially expressed between SSc and DCM myocardium. Mitochondrial energy metabolism pathways, including oxidative phosphorylation, fatty acid β-oxidation and the tricarboxylic acid cycle, were markedly suppressed in SSc. Indirect comparison with NF myocardium confirmed reciprocal regulation of mitochondrial metabolism and suggested enhanced autophagy. Cell deconvolution revealed enrichment of M1-like macrophages in SSc myocardium. LC3 immunostaining and TEM revealed increased autophagic vacuoles, lipid droplet accumulation and ischaemia-like ultrastructural alterations.ConclusionsSSc myocardium exhibits metabolic reprogramming characterised by mitochondrial dysfunction and enhanced autophagy, accompanied by macrophage activation.

ABSTRACTBackgroundAutophagy is an evolutionarily highly conserved process and plays an important role in cellular homeostasis and metabolism. In obesity and obesity‐associated insulin resistance, both enhanced and suppressed autophagy have been observed in adipose tissues (AT).Objective and MethodsWe report on a panel of 7 autophagy markers critically involved in mammalian cellular autophagy in a cohort of patients with obesity (n = 40; mean BMI 54.8 ± 6.7 kg/m2) versus lean individuals (n = 10; mean BMI = 22.7 ± 1.7 kg/m2), during 3T3‐L1 adipocyte differentiation in vitro and in mature 3T3‐L1 adipocytes in response to insulin stimulation under physiologic versus high‐glucose conditions in vitro.ResultsAutophagy markers exhibited a differential expression pattern in subcutaneous AT in patients with obesity as compared to patients with normal weight: LAMP2, ATG5, MAP1LC3B and SIRT1 were reduced in patients with obesity, while LAMP1 and SIRT6 were increased. SIRT1 was reduced in visceral versus subcutaneous AT and in patients with obesity with type 2 diabetes mellitus. ATG5 expression in subcutaneous AT correlated positively with systemic HDL cholesterol levels, and LAMP2 and MAP1LC3B expression in subcutaneous AT correlated positively with systemic pro‐inflammatory Meteorin‐like protein levels (METRNL). LAMP1, LAMP2, BECLIN1, ATG5 and MAP1LC3B increased during 3T3 adipocyte differentiation in vitro, while SIRT1 expression decreased. LAMP2 and MAP3LC3B increased in response to insulin stimulation in mature adipocytes under physiological glucose conditions, while LAMP2 expression increased under high‐glucose conditions. Insulin stimulation reduced SIRT1 expression in mature adipocytes under high glucose conditions. Metformin treatment increased BECLIN1 and ATG5 expression in mature adipocytes.ConclusionsThese observations indicate that adipocyte differentiation, glucose and insulin stimulation, pharmacological interventions and local and systemic inflammation associated with obesity differentially impact on individual autophagy markers both on a cellular and systemic level, potentially explaining the inconsistent effects in patients with obesity reported in the literature.

HighlightsWhat are the main findings?•High expression of GDF15 is closely associated with an obesity-resistant phenotype and contributes to alleviating obesity-induced renal ectopic lipid deposition and kidney injury.•Knockdown of GDF15 receptor GFRAL significantly exacerbated the obese phenotype in mice, accompanied by a reduction in autophagy levels, thereby leading to increased renal lipid deposition and aggravated kidney injury.What are the implications of the main findings?•GDF15 possesses the dual effects of reducing body weight and protecting the kidneys, holding promise as a potential therapeutic target for obesity and its associated nephropathy and providing a new strategy for its clinical prevention and treatment.Objectives: Obesity precipitates excessive lipid accumulation within the kidney, culminating in ectopic lipid deposition that compromises target organ function through lipotoxicity. Given the pivotal role of GDF15 in lipid metabolism, this study aims to determine whether GDF15 can ameliorate ectopic lipid deposition and mitigate the resulting renal injury. Methods: C57BL/6J mice were used to establish a high-fat diet-induced obesity model. Based on Lee’s index, the mice were categorized into a diet-induced obesity group and an obesity-resistant group. Subsequently, the diet-induced obesity group received an injection of AAV-shGFRAL to knock down the GFRAL receptor. Results: In obesity resistant mice, ectopic lipid deposition in the kidneys was markedly reduced, accompanied by decreased expression of the renal injury marker KIM-1 and significantly elevated levels of GDF15. Modulation of the GDF15-GFRAL axis demonstrated that reduced autophagy levels led to increased lipid accumulation and exacerbated renal injury. Conversely, GDF15 activates the AMPK/SIRT1 signaling pathway to promote cellular autophagy, thereby mitigating renal damage induced by ectopic lipid deposition. Consistent with this mechanism, the suppression of autophagy results in the aggravation of renal injury caused by ectopic lipid accumulation. Conclusions: GDF15 ameliorates renal injury induced by ectopic lipid deposition in the kidney primarily through activation of autophagy via the AMPK/SIRT1 signaling pathway.

Endometriosis, a common chronic gynecological disorder, involves cellular autophagy and inflammatory processes in its pathogenesis. However, the specific regulatory mechanisms of autophagy and inflammation in endometriosis remain unknown. In this research, the molecular mechanisms driving the progression of endometriosis are investigated. Through a combination of in vivo and in vitro experiments, this study examines the levels of autophagy and inflammation, as well as the regulatory relationship between SIRT1/FOXO1 and these biological processes. In the in vivo experiments, we successfully established a rat model of endometriosis. The experimental subjects were then divided into a sham-operated group and a model group. Eutopic endometria from the sham group and both eutopic endometria and ectopic lesions from the model group were collected for analysis. The levels of SIRT1, FOXO1, TLR4, NF-κB, and autophagy were assayed through Western blot, PCR, and immunofluorescence experiments. In in vitro experiments, human endometriotic 12Z cells were subjected to FOXO1 inhibition, FOXO1 activation, and SIRT1 suppression to explore the regulatory effect of SIRT1/FOXO1 on cell autophagy and its relationship with endometriosis pathogenesis. The levels of SIRT1, FOXO1, TLR4, NF-κB, and autophagy were assayed through Western blot, PCR, and immunofluorescence experiments. Migration assay, CCK-8 and Transwell assay were used to detect the migration, proliferation, and invasion ability of 12Z. The findings demonstrated that autophagy was markedly upregulated in endometriosis in in vivo experiments. Further analysis indicated that this might be due to the downregulation of SIRT1 levels and the activation of FOXO1. Furthermore, the TLR4-mediated inflammation was significantly enhanced. In vitro experiments further confirmed that autophagy and inflammatory levels in endometriosis cells can be robustly upregulated by activating FOXO1, thus improving their migration, proliferation, and invasion abilities. After SIRT1 suppression, we observed that low SIRT1 levels could increase autophagy by upregulating FOXO1, while activating the TLR4/NF-κB-mediated inflammation, significantly enhancing the implantation, proliferation, and invasion of endometrial cells at the ectopic sites. Based on the in vivo and in vitro results, we found that lower SIRT1 levels could promote the migration, proliferation, and invasion of endometriosis cells by modulating FOXO1 and activating cellular autophagy and the TLR4/NF-κB-mediated inflammation, ultimately accelerating disease progression. By focusing on the SIRT1/FOXO1 axis, this study provides new insights into the pathogenesis of endometriosis and identifies promising treatment targets for future treatment.