关键词： Trigeminal neuropathy   RAGE antagonists   Macrophage   Microglia   Sp5C   ATF3   Activating transcription factor 3   ANOVA   Analysis of variance   CFA   Complete Freund’s adjuvant   DAMP   Damage-associated molecular pattern   dNTPs   Deoxynucleotides   dIoN-CCI   Distal infraorbital nerve chronic constriction injury   GAPDH   Glyceraldehyde-3-phosphate dehydrogenase   HMGB1   High mobility group box-1   IHC   Immunohistochemistry   IoN   Infraorbital nerve   IL-6   Interleukin-6   IL-1β   Interleukin-1β   LPS   Lipopolysaccharide   Me5   Mesencephalic trigeminal nucleus   PTTN   Post-traumatic trigeminal neuropathy   Pr5   Principal sensory trigeminal nucleus   qPCR   Quantitative Real-Time PCR   RAGE   Receptor for advanced glycation end-products   Sp5C   Spinal trigeminal nucleus caudalis   SD   Standard deviation   TNF-α   Tumor necrosis factor-α   TLR4   Toll-like receptor 4   TG   Trigeminal ganglion  

摘要： Post-traumatic trigeminal neuropathy (PTTN) is a debilitating sensory disorder resulting from trigeminal nerve damage for which effective therapies are lacking.   Pretreatment with neutralizing antibodies against high mobility group box-1 (HMGB1), a damage-associated molecular pattern (DAMP), has shown promise in preventing PTTN development; however, the specific receptor mediating HMGB1-driven neuropathic pain in this context remains undefined. This study investigates the role of the receptor for advanced glycation end-products (RAGE), a key HMGB1 receptor, in PTTN pathogenesis using a distal infraorbital nerve chronic constriction injury (dIoN-CCI) model in both male and female mice.   Following dIoN-CCI surgery, mice received two doses of a RAGE antagonist. Pain-like behavior was assessed by measuring facial grooming time, responses to cold stimulation, and mechanical sensitivity. The accumulation of macrophages, as well as the number and volume of microglia, were analyzed by immunohistochemical staining of the injured nerves and the spinal trigeminal nucleus caudalis (Sp5C). dIoN-CCI significantly increased pain-like behaviors in both sexes. Critically, prophylactic treatment with RAGE antagonists significantly attenuated all measured pain responses, regardless of sex. Furthermore, RAGE antagonism markedly reduced macrophage infiltration around the injured nerve and inhibited microglial activation in the Sp5C. These findings provide the first direct evidence that RAGE is crucial in PTTN pathogenesis driven by HMGB1 and demonstrate that RAGE antagonism effectively prevents PTTN development in both sexes. Consequently, early prophylactic treatment targeting RAGE presents a novel and promising strategy for patients at high risk of trigeminal nerve injury.

关键词： Hepatocellular carcinoma   Ferroptosis   Lipid ROS   ATF3   ALOXE3  

摘要： Background Ferroptosis, a novel type of regulated cell death driven by iron-dependent lipid peroxide accumulation, represents a promising therapeutic strategy for aggressive cancers. However, the molecular mechanism of ferroptosis in hepatocellular carcinoma (HCC) remains elusive. Methods RNA sequencing (RNA-seq) identified Activating transcription factor 3 (ATF3) as a key regulator of ferroptosis susceptibility. CCK8 assays, flow cytometry, cell migration assays, colony formation assays, and nanotransmission electron microscopy were performed to investigate the effects of ATF3 in vitro, while subcutaneous xenograft models in nude mice were established to evaluate the biological roles of ATF3 in vivo. Chromatin immunoprecipitation and dual-luciferase assays were conducted to reveal the underlying mechanism of ATF3. Results RSL3-treated HCC cells exhibited characteristic ferroptotic features including elevated lipid peroxidation, mitochondrial shrinkage, and membrane condensation. Clinically, ATF3 expression was significantly higher in adjacent non-tumor tissues compared to HCC tissues and correlated with favorable prognosis, serving as an independent prognostic biomarker. Mechanistically, RSL3-induced ROS accumulation activates ERK/JNK signaling, which upregulates ATF3 expression. As a transcription factor, ATF3 directly binds the ALOXE3 promoter to drive its transcription and subsequent PUFA biosynthesis, thereby sensitizing cells to ferroptosis. Notably, ATF3-overexpressing tumor xenografts showed enhanced sensitivity to RSL3 and significantly enhanced sorafenib's antitumor efficacy. Conclusion RSL3-induced ROS accumulation activated ERK/JNK signaling, upregulating downstream ATF3 expression. ATF3 transcriptionally activated ALOXE3, promoting PUFA synthesis to enhance ferroptosis susceptibility in HCC and overcome sorafenib resistance.

关键词： ATF3   CCDC86   ERK/MAPK signaling   Glioma   Glycolysis  

摘要： Glioma, an aggressively malignant brain tumor with a poor prognosis, comprises nearly 50% of all primary malignant brain tumors. Despite its significance in other cancers, the role of coiled-coil domain containing 86(CCDC86) in glioma remains largely unexplored. Our study revealed a significant up-regulation of CCDC86 expression in glioma tissues, correlating notably with patient age, tumor recurrence, and pathological grade. Moreover, elevated CCDC86 level was associated with a worsened prognosis among glioma patients. Functional assays demonstrated that CCDC86 knockdown attenuated glioma cell proliferation and migration while inducing apoptosis and cell cycle arrest in vitro and inhibited tumorigenesis in vivo . Furthermore, ATF3 emerged as a downstream target gene of CCDC86, as its knockdown could counteract the oncogenic effects induced by CCDC86 overexpression in glioma cells. Mechanistically, CCDC86 promoted the transcriptional regulation of ATF3 by BHLHE40 through interaction with it, stabilizing the expression of ATF3. Additionally, our investigation unveiled a potential mechanism whereby CCDC86 activated the ERK signaling pathway through ATF3, thus influencing glycolysis to drive tumor progression. In conclusion, our study highlights the pivotal role of CCDC86 in glioma progression, suggesting its potential as a therapeutic target for the development of novel glioma treatments.

关键词： ATF3   Calycosin-7-O-β-D-glucoside   Cerebral ischemia/reperfusion injury   Cuproptosis   GSK-3β   SLC31A1  

摘要： Cuproptosis is a type of copper-dependent cell death, yet whether it participates in neuronal death during cerebral ischemia/reperfusion injury (CI/RI) has not been thoroughly investigated. As one of the characteristic components of Astragali radix, calycosin-7-O- β -D-glucoside (CG) exhibits potential pharmacological benefits on CI/RI, but its specific regulatory mechanisms are still insufficiently understood. This study was intended to illustrate the incidence of cuproptosis in CI/RI and to explore the mechanism of CG in mitigating CI/RI. CI/RI was simulated in vitro and in vivo by establishing the oxygen-glucose deprivation/reoxygenation (OGD/R) model of SH-SY5Y cells and the middle cerebral artery occlusion/reperfusion (MCAO/R) model of Sprague-Dawley rats. The results demonstrated that CI/RI induced the onset of cuproptosis accompanied by the expression changes of cuproptosis-associated proteins (FDX1, DLAT, LIAS, and ACO2), intracellular copper enrichment, and depletion of the endogenous copper chelator GSH. The cuproptosis inhibitor ATTM ameliorated cuproptosis, and similarly, CG inhibited cuproptosis, as evidenced that CG significantly boosted OGD/R cell survival, promoted neuronal cell growth, attenuated nerve damage in MCAO/R rats, and reversed the abnormal changes in cuproptosis proteins. Further studies showed that the inhibition of cuproptosis by CG was achieved through inhibition of the GSK-3β/ATF3/SLC31A1 pathway. Either CG or GSK-3β inhibitor LiCl upregulated p-GSK-3β protein expression, inducing a decrease in ATF3 protein expression, which in turn led to a downregulation of SLC31A1 protein expression, a decrease in copper accumulation, and an inhibition of the occurrence of cuproptosis. Investigation of cuproptosis offered therapeutic strategies for CI/RI and other diseases associated with copper disorders.

关键词： Neuroprotection   Cerebral ischemic stroke   Ferroptosis   ATF3   Idebenone analog  

摘要： Ischemic stroke is a leading cause of morbidity and mortality worldwide. Despite its widespread impact, effective treatments remain limited. Idebenone (IDE) has shown promise in promoting recovery following stroke;however, its efficacy during the acute phase of ischemic stroke remains suboptimal. Therefore, the development of novel IDE analogs is of significant importance for improving outcomes in ischemic stroke treatment. Herein, we designed and synthesized a series of novel IDE analogs (1a-1g). An oxygen-glucose deprivation/reoxygenation (O/R) cell model and middle cerebral artery occlusion (MCAO) animal model were employed to investigate the effects of these drugs in vitro and in vivo, respectively. 1c was identified as the most promising candidate owing to its potent neuroprotective effects. Experimental analyses demonstrated that treatment with 1c reduced infarct size following ischemic injury and enhanced neuronal survival. 1c further alleviated neurodegeneration following ischemia/reperfusion (I/R) injury by inhibiting ferroptosis. Furthermore, our findings revealed that the expression of activating transcription factor 3 (ATF3) was upregulated after I/R injury, but markedly suppressed upon treatment with 1c. ATF3 overexpression downregulates the expression of SLC7A11 and GPX4, which are critical regulators of ferroptosis, thereby facilitating ferroptosis following I/R injury. Overall, our findings highlight the therapeutic potential of 1c in ischemic stroke and provide insight into the underlying mechanisms of its neuroprotective effects.

关键词： Sarcopenia   Ferroptosis   ATF3   PI3K/ Akt pathway  

摘要： Purpose: Sarcopenia, characterized by progressive skeletal muscle loss and weakness, has unclear pathogenesis and lacks targeted therapies. Emerging evidence implicates ferroptosis in sarcopenia progression, though its regulatory mechanisms remain undefined. This study investigates the ferroptosis-sarcopenia interplay, identifies core regulators and elucidates their molecular basis. Methods: The experimental design combined in vivo and in vitro approaches using SAMP8 mice and C2C12 myoblast. Sarcopenia phenotypes were systematically characterized through functional assessments of mice, histomorphological analysis of gastrocnemius muscle, and quantification of iron deposition. Bioinformatics cross-analysis was performed by intersecting the sarcopenia-related gene expression dataset (GSE175495) with ferroptosis-associated genes from the FerrDb database, identifying ATF3 as a hub gene. Validation was conducted through Western blot (WB) and quantitative real-time PCR (qPCR). For mechanistic exploration, ferroptosis was induced in C2C12 cells using ferric ammonium citrate (FAC, 500 mu M, 48 h), followed by lentivirus-mediated ATF3 overexpression. The regulatory role of ATF3 in ferroptosis was assessed via reactive oxygen species (ROS) assay, malondialdehyde (MDA) quantification, and FerroOrange fluorescent probe for intracellular iron detection. Transcriptome sequencing of ATF3-dysregulated cell lines was performed, and GO/ KEGG enrichment analyses were applied to identify critical signaling pathways. Functional validation was further conducted using pathway-specific inhibitors. Results: Aged SAMP8 mice exhibited hallmark sarcopenia characteristics including 31 % reduction in grip strength, 42 % decrease in muscle fiber cross-sectional area, and 2.1-fold elevation in intramuscular iron content. Molecular analysis revealed age-dependent downregulation of ATF3 expression (57 % protein decrease, 63 % mRNA reduction). ATF3 overexpression in C2C12 cells significantly attenuat

关键词： HCC therapeutics   DMC   Oxidative stress   Fe-S cluster   Bioenergetics metabolism   ATF4/ATF3/CHOP axis  

摘要： Demethoxycurcumin (DMC) has demonstrated remarkable anti-cancer properties across various tumor types. However, its efficacy in hepatocellular carcinoma (HCC) has not yet been established. Interestingly, the type of cell death induced by DMC was distinct from traditional pathways such as apoptosis, ferroptosis, necroptosis, pyroptosis, cuproptosis, and autophagy, indicating a need for further detailed exploration of its mechanisms. Our investigations revealed that DMC treatment led to a marked increase in oxidative stress, as evidenced by the accumulation of reactive oxygen species (ROS). This increase in ROS disrupted the iron-sulfur (Fe-S) cluster, impairing mitochondrial respiration. Additionally, ROS activation markedly reduced the rate of aerobic glycolysis by interfering with key glycolytic enzymes. The resultant inhibition of these bioenergetic pathways led to a significant depletion of ATP. Moreover, DMC activated the ATF4/ATF3/CHOP signaling axis, and the reduction of CHOP levels mitigated the cytotoxic effects of DMC in HCC cells. In conclusion, DMC initiated oxidative stress, which disrupted bioenergetic metabolism and activated the ATF4/ATF3/CHOP signaling pathway, ultimately leading to cell death in HCC cells. These findings suggest that DMC might have a potential role in the treatment of HCC. However, further validation through in vivo models and clinical trials is required to confirm the therapeutic potential of DMC against HCC. Moreover, more work is still needed to further explore the specific form of cell death induced by DMC.

关键词： 转录激活因子3   敲除   过表达   小鼠精母细胞   炎性因子  

摘要： 旨在探究转录激活因子3(activating transcription factor3,ATF3)表达变化是否与小鼠精母细胞(GC-2spd,GC2)炎性反应相关联,为雄性生殖损伤类疾病发生机制提供见解。将ATF3干扰载体及过表达载体分别转染入GC2,用CCK-8试剂盒检测GC2活力,用实时荧光定量PCR(RT-qPCR)检测GC2的ATF3、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、核因子-κB(nuclear factor kappa-B,NF-κB)、C-C基序趋化因子配体2(C-C motif chemokine ligand 2,CCL2)、C-C基序趋化因子受体2(C-C motif chemokine receptor 2,CCR2)的mRNA表达量;用Western blot检测GC2细胞的ATF3、NF-κB、白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子受体2(tumor necrosis factor receptor 2,TNF-R2)的蛋白表达量。结果发现,过表达ATF3的GC2细胞活力降低,CCL2、CCR2的mRNA表达量显著升高,NF-κB的mRNA和蛋白表达量升高,TNF-R2的蛋白表达量升高。由于ATF3在GC2细胞含量很低,所以GC2细胞敲除ATF3后CCL2、CCR2的mRNA表达量,TNF-α、NF-κB的mRNA和蛋白表达量及TNF-R2的蛋白表达量无显著改变。ATF3通过调控CCL2/CCR2/NF-κB信号通路促进炎性因子表达从而损伤GC2细胞。

关键词： Atherosclerosis   Epidemiology   High-density lipoprotein   Inflammation   Oxidation   Reverse cholesterol transport   Thrombosis   ABCA1   ATP-binding cassette transporter A1   ABCG1   ATP-binding cassette G1   ApoA1   apolipoprotein A1   ApoA2   apolipoprotein A2   ApoCIII   apolipoprotein CIII   ApoE   apolipoprotein E   ApoM   apolipoprotein M   ASCVD   atherosclerotic cardiovascular disease   ATF3   activating transcription factor-3   CAD   coronary artery disease   CE   cholesterol ester   CEC   cholesterol efflux capacity   CHD   coronary heart disease   cIMT   carotid intima media thickness   CKD   chronic kidney disease   CETP   cholesteryl ester transfer protein   CAC   coronary artery calcium   EL   endothelial lipase   eNOS   endothelial nitric oxide synthase   ESRD   end stage renal disease   HDL   high-density lipoprotein   HL   hepatic lipase   IL-1   interleukin-1   IL-6   interleukin-6   IL-10   interleukin-10   IL-12   interleukin-12   iNOS   inducible nitric oxide synthase   LCAT   lecithin cholesterol acyltransferase   LDL   low-density lipoprotein   LDL-R   low-density lipoprotein receptor   LOX-1   lectin-like oxidized low-density lipoprotein receptor 1   LPL   lipoprotein lipase   Lp-PLA2   lipoprotein-associated phospholipase A2   LPS   lipopolysaccharide   LRP   LDL receptor–related protein   LTA   lipoteichoic acid   microRNA   micro ribonucleic acid   MCP-1   monocyte chemoattractant protein-1   MPO   myeloperoxidase   NET   neutrophil extracellular trap   NO   nitric oxide   NOX   NADPH oxidase   NF-KB   nuclear transcription factor kappa B   PON-1   paraoxonase-1   PAF-AH   platelet activating factor acetylhydrolase   PAI-1   plasminogen activator inhibitor-1   PDGFRβ   platelet-derived growth factor receptor β   PGI2   prostaglandin I   2   (aka prostacyclin)   PL   phospholipid   PLTP   phospholipid transfer protein   PTX3   pentraxin-related protein 3   RA   rheumatoid arthritis   ROS   reactive oxygen species   RNS   reactive nitrogen species   RCT   reverse cholesterol transport   S1P   sphingosine-1-phosphate   S1PR3   sphingosine-1-phosphate receptor 3   SAA1   serum amyloid A   SDA   symmetric dimethylarginine   SLE   systemic lupus erythematosus   SR-B1   scavenger receptor class B type 1   STAT3   signal Transducer and Activator of T  

摘要： In contrast to low-density lipoproteins which are atherogenic, high-density lipoproteins (HDL) have been conceptualized as beneficial modulators of adverse pathophysiological phenomena along arterial walls. The HDLs are characterized by highly complex and varied molecular cargoes that include apoproteins, enzymes, microRNAs, bioactive lipids and phospholipids, components of complement, and immune factors, among others. These cargo components determine its functionality. Despite the findings of Mendelian inheritance studies which suggest that HDL is not causal in the pathway for atherogenesis, experiments with HDLs show that it can drive reverse cholesterol transport and antagonize inflammation, oxidation, thrombosis, platelet aggregation, endothelial progenitor cell mobilization, potentiate immunity, foster communication between different cell and tissue types, and function as a crucial apoprotein donor amongst the various lipoproteins. These functions are understandably viewed as beneficial and antagonize pathophysiology. Secondary to the complexity of its proteome and lipidome, HDL functionality is profoundly responsive to the metabolic and genetic backgrounds of individuals. Even its size and lipidation status can influence its functionality. As part of the acute phase response, critical antioxidative moieties can be replaced by such acute phase reactants as serum amyloid A and pro-oxidative enzymes. The functionality of HDL is influenced by chronic kidney disease, coronary artery disease, acute myocardial infarction, obesity, insulin resistance, metabolic syndrome, diabetes mellitus, and cancer. Herein we describe many of the alterations in HDL constitution and the resulting changes in functional capacity that can be observed. A unifying theme characterizing these disease states is that they all heighten systemic inflammatory tone and potentiate a pro-oxidative state. These changes clearly associate with profound changes in the functionality and behavior of HDL

关键词： DHA   EPA   PUFA-PLs   Ferroptosis   Hypoxia   SLC7A11   Gastrointestinal mucosa   4-HNE   4-hydroxynonenal   ALOX5   5-lipoxygenase   ANOVA   one-way analysis of variance   ATF3   activating transcription factor 3   cAMP   cyclic adenosine monophosphate   CCK-8   cell counting kit-8   DHA   docosahexaenoic acid   DMEM   Dulbecco's modified Eagle’s medium   DMSO   dimethyl sulfoxide   EPA   eicosapentaenoic acid   FBS   fetal bovine serum   Fer-1   ferrostatin-1   GSH   glutathione   GPX4   glutathione peroxidase 4   H&E   hematoxylin and eosin   HIEC   normal human small intestinal epithelial cell line   HIF   hypoxia-inducible factor   IL-6   interleukin-6   Lip-1   liproxstatin-1   LPO   lipid hydroperoxide   MDA   malondialdehyde   n-3   Omega-3   NGEC   Normal human gastric epithelial cell line   NOX4   NADPH oxidase 4   PCs   phosphatidylcholines   PEs   phosphatidylethanolamines   PGs   phosphatidylglycerols   PIs   phosphatidylinositols   PKA   protein kinase A   PLs   phospholipids   PSs   phosphatidylserines   PUFAs   polyunsaturated fatty acids   ROS   reactive oxygen species   SEM   standard error of the mean   TEM   transmission electron microscopy  

摘要： Hypoxia, resulting from environmental factors or diseases, can disrupt the gastrointestinal mucosal barrier. Our previous study demonstrated that hypoxia induced ferroptosis in the gastric and small intestinal mucosa by upregulating ALOX5, NOX4, and polyunsaturated fatty acid-containing phospholipids (PUFA-PLs). The impact of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) on ferroptosis is currently a subject of debate. While DHA and EPA upregulate SLC7A11 expression, mitigating lipid peroxidation, they also elevate PUFA-PL levels, exacerbating it. This study investigated the effects and underlying mechanisms of DHA and EPA supplementation on gastric and small intestinal mucosal ferroptosis under normoxic and hypoxic conditions in vitro and in vivo. Under normoxia, DHA and EPA upregulated SLC7A11 expression through the cAMP/PKA/ATF3 pathway, thereby enhancing cellular resistance to lipid peroxidation associated with increased PUFA-PL levels and preventing ferroptosis. In contrast, under hypoxia, DHA and EPA exacerbated ferroptosis by further increasing PUFA-PL levels, which, in combination with hypoxia-induced ALOX5 and NOX4 expression, resulted in excessive lipid peroxidation that overwhelmed the protective mechanisms mediated by SLC7A11 upregulation. These findings indicate that DHA and EPA exacerbate hypoxia-induced ferroptosis in gastric and small intestinal mucosa. Therefore, individuals at risk of hypoxia should carefully consider the potential risks associated with DHA and EPA intake.