The FASEB Journal最新文献

Essential meiotic structure-specific endonuclease 1 (EME1) is integral to the maintenance of genomic stability in various cancers. However, its biological role and expression profile of this molecule in nasopharyngeal carcinoma (NPC) remain to be explored. In this study, we found that EME1 was overexpressed in NPC specimens compared to adjacent noncancerous tissues and was correlated with poorer overall survival outcomes. Furthermore, EME1 knockdown significantly inhibited the proliferation and migration of NPC cells in vitro and in vivo, with a corresponding sensitization to either camptothecin (CPT) or olaparib, evidenced by a further suppression of proliferation upon drug treatment. Notably, silencing EME1 significantly increased the sensitivity of NPC cell lines to CPT by enhancing ATM-CHEK2 phosphorylation and inducing nuclear abnormalities. Collectively, our findings suggest that combining EME1 modulation with agents such as CPT or olaparib could be an effective treatment strategy for NPC patients.

Radiotherapy activates both the PI3K/AKT pathway and autophagy in cervical cancer, contributing to radioresistance. To address this, EM2, a dual AKT/autophagy inhibitor, was investigated for its potential to enhance radiosensitivity. RNA-Seq, Western blot, qRT-PCR, and transmission electron microscopy were employed to analyze PI3K/AKT and autophagy pathways following irradiation, while CCK8, clone formation, and flow cytometry assays evaluated proliferation, apoptosis, and cell cycle effects. KEGG and GSEA analyses confirmed irradiation-induced activation of the PI3K/AKT pathway. Both PI3K and autophagy inhibitors significantly improved efficacy, whereas EM2 suppressed AKT pathway activation and autophagy, synergistically inducing G2/M phase arrest, and increasing apoptosis. In vivo experiments using a nude mouse xenograft model demonstrated that EM2 combined with irradiation effectively suppressed tumor growth, PI3K/AKT activation, and autophagy without significant toxicity. These results underscore EM2 as a promising therapeutic agent to overcome radioresistance by simultaneously targeting the PI3K/AKT pathway and autophagy.

Exposure to small independent space (SIS) causes stress-related behavioral and neural abnormalities, but the time-dependent mechanisms and gut microbiota-hippocampus interactions remain unclear. To investigate the time-dependent effects of acute and chronic restraint stress (CRS) on mouse behavior, neuroendocrine function, gut microbiota, and hippocampal activity. Behavioral assessments were conducted using the open field test and tail suspension test. Meanwhile, enzyme-linked immunosorbent assay (ELISA) was applied to measure neurotransmitters, hypothalamic–pituitary–adrenal (HPA) axis markers, and inflammatory factors; 16S rRNA sequencing was used for gut microbiota analysis; liquid chromatography–tandem mass spectrometry (LC–MS/MS) for metabolite identification; and immunofluorescence staining for hippocampal structure observation. Acute SIS induced depression-like behaviors. Chronic SIS showed peak despair at 4 days (d), followed by persistent depression at 8 d. Norepinephrine (NE) increased while 5-hydroxytryptamine (5-HT) decreased; NE depletion at 4 d coincided with reduced hippocampal glucocorticoid receptor (GR) expression. Both acute (2 h) and chronic (4 d) SIS reduced gut microbiota diversity and disrupted tyrosine metabolism. Hippocampal tests showed neuronal compensation at 6 h post-acute stress and neuronal apoptosis with glial dysfunction after 4 d of chronic stress. SIS damages via the “gut microbiota-tyrosine metabolism-hippocampus axis,” with 4 d of chronic exposure as a critical decompensation point, offering a time-specific intervention target.

Mechanical force induces a series of biological responses such as inflammation in force-loaded tissues and cells. The periodontal ligament (PDL) fibroblasts act as vital sensors and transducers in response to mechanical loading within periodontium. Studies have shown that PDL fibroblasts also participate in mediating periodontal inflammatory responses under physiological or pathological conditions. Mitophagy is a selective form of autophagy that eliminates damaged or dysfunctional mitochondria to maintain cellular health. It plays a vital role in inflammation alleviation, cell survival, and tissue homeostasis. However, whether mitophagy is involved in mechanical force-related inflammation and the precise mechanisms remain unclear. In addition, the elucidation of the interplay between mitophagy and periodontal inflammation during mechanical loading is of great significance for maintaining periodontal homeostasis under systemic conditions. In our study, we first focused on validating the crosstalk between mitophagy and inflammation in PDL fibroblasts under mechanical loading and aimed to elucidate the upstream regulatory role of adenosine monophosphate-activated protein kinase (AMPK). Moreover, based on both in vivo and in vitro experiments, we found that high glucose conditions exacerbated inflammation by suppressing mitophagy. Additionally, targeted activation of AMPK enhanced mitochondrial turnover through mitophagy, thereby disrupting proinflammatory cascades and offering a promising strategy for inflammation resolution in periodontal diseases, especially those combined with diabetic conditions.

Diabetic kidney disease (DKD) is the primary cause of end-stage renal disease globally, yet reliable biomarkers for its diagnosis and progression assessment are lacking. This study employed artificial intelligence techniques, including weighted gene co-expression network analysis (WGCNA) and machine learning, to identify crucial genes associated with DKD. Validation was conducted using online databases such as Nephroseq and KIT, alongside biological samples from human serum, urine, peripheral blood mononuclear cell (PBMC) mRNA, kidney tissues, DKD rat models, and high glucose-treated HK-2 cells. Statistical analyses evaluated the correlations. The study revealed that C-X-C motif chemokine ligand 3 (CXCL3) was markedly upregulated in the serum and urine of DKD patients compared to healthy controls and those with type 2 diabetes mellitus, primary glomerulonephritis (e.g., IgA nephropathy, membranous nephropathy, minimal change disease). Immunohistochemistry showed significantly higher CXCL3 in both the glomeruli and tubulointerstitium of DKD patient kidneys than in those from controls. Elevated CXCL3 mRNA levels were also noted in PBMCs from DKD patients, STZ-induced DKD rat kidneys, and high glucose-treated HK-2 cells. Furthermore, urinary CXCL3 protein levels positively correlated with the pathological grade, serum blood urea nitrogen (BUN), serum creatinine, and HbA1c percentage, while inversely correlating with estimated glomerular filtration rate (eGFR) in DKD patients. Mechanically, high glucose stimulation significantly upregulates the expression of inflammatory factors (including IL-6 and IL-1β) and fibrosis markers (α-SMA and CTGF) in HK-2 cells overexpressing CXCL3. Conversely, CXCL3 knockout in HK-2 cells led to substantial downregulation of these inflammatory and fibrotic markers in the same high glucose conditions. Elevated CXCL3 levels in the serum, urine, and kidney tissues, alongside increased mRNA in PBMCs, suggest its potential as a biomarker for diagnosing and monitoring DKD. Correlations of urinary CXCL3 with disease severity indicators further support its diagnostic and prognostic utility. Mechanically, CXCL3 promotes inflammation and fibrosis in DKD.

Silicosis, an occupational pulmonary fibrosis caused by silica dust exposure, lacks effective treatments. This study investigates the therapeutic potential and mechanism of methyl gallate (MG), a natural polyphenol, in silicosis fibrosis. A silica-induced silicosis mouse model and TGF-β1-stimulated human lung fibroblasts were employed. MG administration significantly ameliorated lung fibrosis in mice, reducing collagen deposition, α-SMA, fibronectin, and TGF-β1 levels. Transcriptomic analysis revealed that MG inhibited fibroblast activation by suppressing cell cycle progression via CDK6-mediated G0/G1 arrest. Mechanistically, MG downregulated MDM4 protein levels, disrupted MDM4-P53 interaction, and activated the P53-P21 pathway, promoting fibroblast apoptosis and cell cycle arrest. Further, Drug Affinity Responsive Target Stability (DARTS) and Cellular Thermal Shift Assay (CETSA) identified hnRNPA2/B1 as MG's direct target. MG inhibited hnRNPA2/B1-mediated MDM4 mRNA translation, thereby reducing MDM4 protein synthesis. Overexpression of MDM4 or knockdown of hnRNPA2/B1 reversed MG's anti-fibrotic effects. These findings highlight MG's novel role in alleviating silicosis fibrosis by targeting the hnRNPA2/B1-MDM4-P53 axis, offering a promising therapeutic strategy for silicosis.

Lonicerae Japonicae Flos (LJF), a traditional Chinese medicine with strong anti-inflammatory and antioxidant effects, was previously shown to suppress ferroptosis in acute lung injury (ALI). This study further aimed to identify its anti-ferroptotic quality markers (Q-markers) and elucidate their mechanisms for ALI therapy. Chemical fingerprints of 22 LJF batches were established, and absorbed constituents were identified through serum pharmacochemistry. Principal component analysis (PCA) and gray correlation analysis (GCA) were applied to link chemical composition with pharmacological effects. Ferroptosis-related indicators were measured in an LPS-induced ALI mouse model after treatment with different extracts. The anti-ferroptotic activity of individual constituents was validated in BEAS-2B cells, leading to the identification of Q-markers. To confirm target engagement, transcriptomic sequencing, protein–protein interaction (PPI) network construction, ferroptosis-target enrichment, real-time quantitative polymerase chain reaction (RT-qPCR), molecular docking, and molecular dynamics (MD) simulations were conducted. 17 candidate constituents with potential anti-ferroptotic and anti-inflammatory activities were identified, of which 9 showed strong correlations with efficacy. Four compounds—protocatechuic acid, loganin, cynaroside, and isochlorogenic acid C—demonstrated significant ferroptosis-inhibitory activity and were designated as Q-markers. Transcriptomic and PPI analyses indicated that these constituents target key ferroptosis-related proteins, including IL-6, Stat3, and Mapk3. Their regulatory effects and binding stability were further validated by RT-qPCR, molecular docking, and MD simulations. LJF exerts anti-inflammatory protection through the synergistic inhibition of ferroptosis by multiple constituents. The identification of protocatechuic acid, loganin, cynaroside, and isochlorogenic acid C as Q-markers provides scientific evidence supporting the therapeutic potential of LJF for ALI.

Non-alcoholic fatty liver disease (NAFLD) is a global health burden characterized by hepatic steatosis and progressive fibrosis, necessitating novel therapeutic strategies. This study investigates the molecular mechanism by which cyclic RGD peptide (cRGD)-modified adipose-derived mesenchymal stem cell (ADMSC)-derived extracellular vesicles (EVs) deliver Angiotensin-(1–7) to attenuate NAFLD-associated liver fibrosis. EVs were isolated from murine ADMSCs via ultracentrifugation, surface-modified with cRGD using EDC/NHS crosslinkers, and loaded with Angiotensin-(1–7) via an ultrasound-assisted method. The therapeutic effects were evaluated in vitro using hepatic stellate cells (LX-2) and in vivo using a high-fat diet (HFD)-induced NAFLD mouse model. Multi-omics analyses (transcriptomics, proteomics, metabolomics) were performed on liver tissues to elucidate underlying pathways. Results demonstrated that cRGD-modified EVs loaded with Angiotensin-(1–7) exhibited excellent biocompatibility and targeted liver accumulation, significantly reducing hepatic lipid accumulation, fibrosis, and serum markers of liver damage (ALT, AST). Mechanistically, Angiotensin-(1–7) activated the Mas receptor, enhancing Akt-Foxo1-dependent autophagy and fatty acid metabolism reprogramming, as confirmed by upregulation of autophagy-related proteins (LC3-II, p62) and downregulation of fibrosis markers (TGF-β1, α-SMA, Collagen I). Multi-omics data revealed enrichment in fatty acid degradation and autophagy pathways, while Mas receptor inhibition abolished these effects. This study establishes that cRGD-modified EVs deliver Angiotensin-(1–7) as a potent strategy to mitigate NAFLD fibrosis through Mas/Akt/Foxo1 signaling, offering a promising therapeutic avenue for metabolic liver diseases.