关键词： adipogenesis   ATF3   USP53   RhoA/ROCK pathway   transcriptional regulation  

摘要： Obesity is a widespread nutritional disorder, leading to a strong predisposition toward adverse health consequences. Activating transcription factor 3 (ATF3), a stress-induced transcription factor, has been documented as a therapeutic target for obesity. The intent of this project was to characterize the detailed role of ATF3 in adipogenesis in the context of obesity and its obscure downstream mechanism. After adipogenic differentiation, RT-qPCR and western blot examined ATF3 and ubiquitin-specific peptidase 53 (USP53) mRNA levels and protein levels. Adipogenesis was identified by Oil Red O staining, triglyceride (TG) levels, and western blot analysis. JASPAR database, ChIP and luciferase reporter assays predicted and validated the transcriptional regulation of USP53 by ATF3. Western blot also examined the protein levels of Ras homolog family member A (RhoA)/Rho-associated coiled-coil kinase (ROCK) pathway-involved proteins. ATF3 mRNA and protein levels were depleted in the differentiated 3T3-L1 adipocytes, and ATF3 elevation hindered the adipogenesis of 3T3-L1 adipocytes. ATF3 suppressed the transcription of USP53 as a transcription factor and lowered USP53 expression. Eventually, USP53 upregulation partially blunted the inhibitory role of ATF3 overexpression in adipogenesis and the RhoA/ROCK pathway. Consequently, ATF3 might transcriptionally inactivate USP53 to repress adipocyte adipogenesis and downregulate the RhoA/ROCK pathway.

关键词： Gasoline exhaust particles   Activating transcription factor 3   Matrix metalloproteinase 1   Nicotinamide adenine dinucleotide phosphate oxidase 4   human umbilical vein endothelial cells   reactive oxygen species  

摘要： Gasoline exhaust particles (GEP) are risk factors for cardiovascular disease. Activating transcription factor 3 (ATF3) is a transcription factor known to form a heterodimer with AP-1 transcription factors for its target gene expression. However, the involvement of ATF3 in GEP-induced gene expression in human umbilical vein endothelial cells (HUVECs) has not been investigated. In this study, we found that GEP, at IC50 value of 59 mu g/ml, induced the expression of ATF3, which led to the expression of matrix metalloproteinase 1 (MMP1) in HUVECs. GEP induce an interaction between c-Jun and ATF3, and c-Jun depletion attenuates GEP-induced MMP1 expression. Depletion of NADPH oxidase 4 (Nox4) suppressed GEP-induced reactive oxygen species (ROS) generation and the subsequent upregulation of ATF3 and MMP1, suggesting that Nox4-derived ROS play a role as upstream regulators of GEP-induced ATF3 expression and MMP1 upregulation. Furthermore, Nox4 depletion attenuated the interaction between ATF3 and c-Jun and their binding to the AP-1 binding site of the MMP1 promoter. Taken together, these findings demonstrate that GEP induce the expression of MMP1 by generating Nox4-dependent ROS, which subsequently increase ATF3 expression and its interaction with c-Jun. This leads to their binding to the promoter region of MMP1 and its transcription. These findings suggest that Nox4-derived ROS and ATF3 are critical for GEP-induced MMP1 expression.

关键词： Myocardial ischemia-reperfusion injury   Melatonin   Ferroptosis   ATF3   GPX4  

摘要： Melatonin (MEL), functioning as a circulating hormone, is important for the regulation of ferroptosis in different health scenarios and acts as a crucial antioxidant in cardiovascular diseases. However, its specific function in ferroptosis related to myocardial ischemia-reperfusion injury (MIRI) remains to be fully elucidated. In our research, we utilized a rat model of MIRI induced by coronary artery ligation, along with a cell model subjected to hypoxia/reoxygenation (H/R). We evaluated relevant genes and proteins by real-time fluorescent quantitative PCR and Western blot analysis. To evaluate myocardial tissue damage and cell injury, we employed cell counting kit-8 assays, flow cytometry, hematoxylin-eosin staining, and 2,3,5-triphenyltetrazolium chloride staining techniques. Our results show that administering MEL notably reduces the concentrations of cTnT, CK-MB, and lactate dehydrogenase in the serum of MIRI rats, mitigates the extent of myocardial infarction, improves the recovery of pathological conditions in myocardial tissues, and reduces the concentrations of Fe2+, malondialdehyde (MDA), and reactive oxygen species (ROS) in the myocardial tissue, while also promoting increased glutathione levels. Moreover, MEL can also restore the reduced viability of H9C2 cells caused by H/R or ferroptosis inducers (RSL3), reduce the cellular content of Fe2+, MDA, and ROS, and inhibit ferroptosis. Mechanistically, MEL promotes the expression of GPX4 by downregulating the expression of ATF3, thereby inhibiting ferroptosis in cardiomyocytes and ultimately alleviating the process of MIRI. Our study demonstrates that MEL ameliorates MIRI by inhibiting ferroptosis.

关键词： Lupus nephritis   Ferroptosis   ATF3   Machine learning  

摘要： Background: The role of ferroptosis in lupus nephritis (LN) is unclear. This study aimed to explore the effects of ferroptosis-related genes in LN through bioinformatics prediction and experimental validation. Methods: Sample data were collected from the GEO dataset and divided into glomeruli and tubulointerstitium. We collected 382 ferroptosis-related genes. The intersection of ferroptosis-related genes with glomeruli and tubulointerstitium data, respectively, was performed. Machine learning methods (including unsupervised cluster typing and random forests) were operated to identify ferroptosis subtyping and ferroptosis important genes in LN. Immune infiltration and functional analysis were performed. The expression of ferroptosis important gene ATF3 was validated in vivo and in vitro. Results: 6 ferroptosis important genes common to glomeruli and tubulointerstitium were screened, including ATF3, CD44, CYBB, JUN, NCF2, and NNMT. ATF3 decreased in the LN group compared to the Control. Silencing ATF3 mitigated LPS/erastin-induced ferroptosis. Functional analysis showed that ATF3 was markedly enriched in the interferon-gamma-mediated signaling pathway, ECM-receptor interaction, and cell adhesion. In glomeruli, T cells regulatory (Tregs) infiltration decreased and Macrophages M1 levels increased with elevated ATF3 expression. Levels of immune cell infiltration were altered in different ferroptosis subtypes of LN glomeruli and tubulointerstitium. Conclusions: Ferroptosis-related ATF3 levels decreased in LN. Inhibition of ATF3 might alleviate LN development by affecting the macrophage M1 and Treg cell infiltration. These implied that ATF3 might be a potential target for developing LN therapeutic strategies.

关键词： Triple-negative breast cancer   Aurovertin B   Dual-specificity phosphatase 1   Activating transcription factor 3   Lung metastasis  

摘要： Triple-negative breast cancer (TNBC) is characterized by high mortality rates, primarily due to its propensity for metastasis. Addressing this challenge necessitates the development of effective antimetastatic therapies. This study aimed to identify natural compounds with potential antimetastatic properties mainly based on the high-throughput phenotypic screening system. This system, utilizing luciferase reporter gene assays combined with scratch wound assays, evaluates compounds based on their influence on the epithelialmesenchymal transition (EMT) marker E-cadherin. Through this approach, aurovertin B (AVB) was revealed to have significant antimetastatic capability. Notably, AVB exhibited substantial metastasis suppression in many TNBC cell lines, including MDA-MB-231, HCC1937, and 4T1. Also, its remarkable antimetastatic activity was demonstrated in vivo via the orthotopic breast cancer mouse model. Further exploration revealed a pronounced association between AVB-induced upregulation of dual-specificity phosphatase 1 (DUSP1) and its inhibitory effect on TNBC metastasis. Additionally, microarray analysis conducted to elucidate the underlying mechanism of the AVB-DUSP1 interaction identified activating transcription factor 3 (ATF3) as a critical transcription factor instrumental in DUSP1 transcriptional activation. This discovery, coupled with observations of enhanced ATF3-DUSP1 expression and consequent reduction in TNBC metastatic foci in response to AVB, provides novel insights into the metastatic mechanisms of TNBC. Significance Statement: This study constructs a high-throughput phenotypic screening system utilizing epithelial-mesenchymal transition marker E-cadherin promoter luciferase reporter gene combined with scratch wound assays. Aurovertin B was revealed to possess significant antimetastatic activity through this approach, which was further demonstrated via in vivo and in vitro experiments. The discovery of the regulatory role of the ATF3-DUSP1 pathway

关键词： aortic aneurysm and dissection   cGAS-STING pathway   DNA stability   micronuclei formation   P21  

摘要： BackgroundSporadic aortic aneurysm and dissection (AAD) is a critical condition characterised by the progressive loss of vascular smooth muscle cells (VSMCs) and the breakdown of the extracellular matrix. However, the molecular mechanisms responsible for the phenotypic switch and loss of VSMCs in AAD are not fully *** and resultsIn this study, we employed a discovery-driven, unbiased approach. This approach encourages us to explore the unknown functions of activating transcription factor 3 (ATF3) rather than merely confirming existing hypotheses, while no assumptions were made about ATF3 prior to the experiments. We ensured the unbiased nature of our assessment by conducting morphological evaluations with two independent observers in a blinded manner. We identified elevated expression of ATF3 in both human sporadic AAD tissues and mouse AAD models. VSMC-specific ATF3 conditional knockout (Atf3 cKO) mice showed notable enlargement, dissection and rupture in both thoracic and abdominal aortic regions after exposure to Ang II. Interestingly, older Atf3 cKO mice exhibited spontaneous aortic dissections and senescence of the aortic wall. Mechanistically, ATF3 deficiency led to the degradation of P21 through ubiquitination. Impaired DNA repair in VSMCs resulted in micronuclei formation in the cytoplasm, activating the cyclicGMP-AMP synthase- stimulator of interferon genes (cGAS-STING) pathway and inducing VSMC phenotypic switching and apoptosis. Finally, both pharmacological complementation of P21 function and knockdown of STING expression alleviated ATF3 deficiency-induced *** study indicates that ATF3 is essential for genomic DNA stability in VSMCs through the P21-cGAS-STING pathway, suggesting that enhancing ATF3 expression in VSMCs could help prevent sporadic *** points ATF3 deficiency led to degradation of P21 through ubiquitination, which abolished the G1 phase arrest. VSMCs had no time window to repair the damaged DNA, leading to g

关键词： breast cancer   H3K27ac   IFI30   tumor-associated macrophages  

摘要： Immunotherapy has transformed cancer treatment, but its effectiveness in breast cancer remains suboptimal. Tumor-associated macrophages (TAMs), a key component of the tumor microenvironment (TME), contribute significantly to immune evasion. In this study, we identified gamma-interferon-inducible lysosomal thiol reductase (IFI30) as a critical regulator of TAM function in breast cancer. IFI30 expression is upregulated in breast cancer via enhanced Histone 3 lysine 27 acetylation (H3K27ac) modification and promotes tumor progression and metastasis in an immune-dependent manner. Mechanistically, IFI30 in breast cancer cells recruits TAMs by activating the ATF3-CCL5 axis. Within macrophages, it promotes M2-like polarization and PD-L1 upregulation, fostering an immunosuppressive TME. Our findings established IFI30 as a promising therapeutic target for disrupting TAM-mediated immune suppression and enhancing breast cancer immunotherapy. © 2025 The Author(s)

关键词： 高血压肾病   细胞衰老   差异表达基因   生物信息学   作用机制  

摘要： 目的 借助生物信息学技术研究细胞衰老在高血压肾病中可能发生的机制及相关信号通路。方法 查找基因表达数据库下载微阵列数据GSE37455,用R软件合并数据并去除批次效应，获取差异表达基因。从细胞衰老基因CellAge数据库获取细胞衰老基因，通过R包“ggVennDiagram”获取与细胞衰老相关差异基因。导入基因相互作用网络String数据库获取蛋白质互作(PPI)网络图，用Cytoscape 3.8.0软件挖掘核心差异基因。用基因本体(GO)、京都基因与基因组百科全书(KEGG)、基因集富集分析(GSEA)对核心差异基因进行功能和通路分析。结果 共获得41个差异基因，包括32个上调基因和9个下调基因。取交集后获得4个与细胞衰老相关差异基因。通过PPI分析和Cytoscape处理获得核心差异基因激活转录因子3(ATF3)。GSEA分析显示，细胞衰老可能通过线粒体解偶联蛋白1产热、Ⅰ型干扰素反应(TYPE-Ⅰ-IFN)的信号传导等途径在高血压肾病中发挥作用。结论 细胞衰老在高血压肾病中可能通过线粒体解偶联蛋白1产热、TYPE-Ⅰ-IFN等途径发生作用，为高血压肾病的研究提供治疗靶点。

关键词： Sjogren's syndrome   Apigenin   Salivary gland epithelial cells   Ferroptosis  

摘要： Background: In Sjogren's ' s syndrome (SS)-an - an autoimmune disease characterized by dry mouth and eyes-salivary - salivary gland epithelial cells (SGECs) undergo ferroptosis, which disrupts their integrity and impairs saliva secretion. Apigenin, a phytoestrogen, is known to activate estrogen signalling and alleviate xerostomia in ovariectomized mice;however, its effect on SGEC survival and function in SS remains unclear. We hypothesized that apigenin alleviates SS symptoms and progression by inhibiting ferroptosis in SGECs and aimed to elucidate the underlying mechanism. Methods: Apigenin (50 mg/kg) was orally gavaged to non-obese diabetic (NOD)/LtJ female mice (SS model);changes in SS functional indicators were analyzed using mRNA sequencing and bioinformatic analyses of sub- mandibular glands. Interferon-gamma (IFN-gamma)-stimulated gamma )-stimulated SGECs were used to model SS in vitro;SGEC activity and aquaporin 5 (AQP5) expression were analyzed. Immunohistochemical staining, transmission electron microscopy, RT-qPCR, western blotting and other methods were used to verify the mechanisms. Results: Apigenin significantly increased salivary secretion and AQP5 expression while inhibiting ferroptosis and immune infiltration in NOD mouse submandibular glands. The oxidative stress gene ATF3 was upregulated and GPX4 was downregulated in NOD mice compared to that in control group (ICR mice);however, apigenin reversed this effect. IFN-gamma gamma treatment downregulated AQP5, SLC7A11, and GPX4 expression while promoting ATF3 expression and ferroptosis, which was mitigated by apigenin. ATF3 knockdown increased SLC7A11 and GPX4 expression, inhibiting SS and ferroptosis. Furthermore, apigenin inhibited ferroptosis in SGECs through ESR1 binding to ATF3. Conclusion: Apigenin alleviates SS by regulating SGEC ferroptosis via the ER alpha-regulated ATF3/SLC7A11 axis, highlighting its therapeutic potential in SS.

关键词： Neuronal oxidative stress   FTO   M(6)A modification   Cognitive dysfunction  

摘要： Excessive exposure to metals in daily life has been proposed as an environmental risk factor for neurological disorders. Oxidative stress is an inevitable stage involved in the neurotoxic effects induced by metals, nevertheless, the underlying mechanisms are still unclear. In this study, we used arsenic as a representative environmental heavy metal to induce neuronal oxidative stress and demonstrated that both in vitro and in vivo exposure to arsenic significantly increased the level of N6-methyladenosine (m6A) by down-regulating its demethylase FTO. Importantly, the results obtained from FTO transgenic mice and FTO overexpressed/knockout cells indicated that FTO likely regulated neuronal oxidative stress by modulating activating transcription factor 3 (ATF3) in a m(6)A-dependent manner. We also identified the specific m6A reader protein, YTHDC1, which interacted with ATF3 and thereby affecting its regulatory effects on oxidative stress. To further explore potential intervention strategies, cerebral metabolomics was conducted and we newly identified myo-inositol as a metabolite that exhibited potential in protecting against arsenic-induced oxidative stress and cognitive dysfunction. Overall, these findings provide new insights into the importance of the FTO-ATF3 signaling axis in neuronal oxidative stress from an m(6)A perspective, and highlight a beneficial metabolite that can counteract the oxidative stress induced by arsenic.