关键词： ATF3   chromatin dynamics   development   enhancers   hepatocyte   liver   NRF2   regeneration   transcription factors  

摘要： The mammalian liver exhibits remarkable regenerative capabilities after injury or resection. Central to this process is the precise modulation of gene expression, driven by changes in chromatin structure and the temporal activation of distal regulatory elements. In this study, we integrated chromatin accessibility and transcriptomic data after partial hepatectomy in mice. We show that the expression of crucial regeneration genes is orchestrated by a diverse array of cis-regulatory elements, including regeneration-specific enhancers and enhancers repurposed from various developmental stages. These enhancers collaborate to activate the transcriptional programs required for hepatocyte priming and proliferation, with their activity initially regulated by the activator protein-1 (AP-1) complex and ATF3, and subsequently by nuclear factor erythroid 2 (NFE2)-related factor 2 (NRF2) during proliferation. Our results also indicate that hepatic regeneration involves the repression of enhancers regulating liver-specific metabolic functions, particularly those involved in lipid metabolism. This study provides a genome-wide atlas of enhancer-gene interactions, offering new insights into the regulatory mechanisms underlying liver regeneration. © 2025 The Author(s)

关键词： Intracerebral hemorrhage   Ferroptosis   Inflammation   BRD4   ATF3  

摘要： BackgroundIntracerebral hemorrhage (ICH), a subtype of hemorrhagic stroke, is associated with high morbidity and mortality. This study aimed to investigate the role and underlying mechanism of bromodomain and extraterminal domain protein 4 (BRD4) in neural injury following *** in vitro ICH model was established by treating SH-SY5Y cells with hemin. BRD4 and activating transcription factor 3 (ATF3) expression levels were altered using short hairpin RNA and pcDNA3.1 expression vectors. Cell viability and death were assessed using Cell Counting Kit-8 (CCK-8) and TdT-mediated dUTP nick-end labeling (TUNEL) assays. mRNA and protein expression levels were quantified by RT-qPCR and western blotting, respectively. Cytokine secretion was measured using enzyme-linked immunosorbent assay (ELISA) kits. Ferroptosis was evaluated based on levels of reactive oxygen species (ROS), superoxide dismutase (SOD), malondialdehyde (MDA), and Fe2(+). Chromatin immunoprecipitation (ChIP) was performed to examine the interaction between BRD4, histone 3 lysine 27 acetylation (H3K27ac), and the ATF3 ***4 expression was upregulated in hemin-treated SH-SY5Y cells, and its knockdown repressed inflammation and ferroptosis induced by hemin. ATF3 expression was also elevated upon hemin stimulation and was identified as a downstream target of BRD4. Mechanistic studies revealed that BRD4 cooperated with H3K27ac to bind the ATF3 promoter, thereby promoting ATF3 transcription. Functional rescue experiments further demonstrated that ATF3 overexpression counteracted the inhibitory effects of BRD4 knockdown on hemin-induced inflammation and ***4 silencing alleviated hemin-induced neuronal ferroptosis and inflammation by regulating ATF3 transcription, suggesting that BRD4 inhibition may represent a potential therapeutic strategy for ICH.

关键词： Toxoplasma gondii   Adverse pregnancy outcomes   Macrophages   Trem2 promoter   ATF3  

摘要： BackgroundApproximately one in three people worldwide have been exposed to Toxoplasma gondii (T. gondii). Primary infection with T. gondii during pregnancy can cause severe complications. Our previous study demonstrated that deficiency of triggering receptor expressed on myeloid cells 2 (Trem2) exacerbates pregnancy-related complications in T. gondii-infected mice. However, understanding the mechanisms by which T. gondii modulates Trem2 expression in macrophages remains an unmet *** mouse pregnancy model of T. gondii infection and an in vitro cellular stimulation model using soluble T. gondii antigens (sTgAg) were used to assess Trem2 expression. Recombinant plasmids containing the full-length Trem2 promoter were constructed to evaluate the effect of sTgAg on promoter activity, followed by the construction of truncated promoter plasmids to identify key regulatory regions. Transcription factors potentially binding to the Trem2 promoter were predicted using PROMO and JASPAR, with ATF3 identified as responsive to sTgAg stimulation via western blot analysis. The binding of ATF3 to the Trem2 promoter was validated by chromatin immunoprecipitation (ChIP) assays. Finally, ATF3 knockdown experiments were performed to determine its role in mediating the inhibitory effect of sTgAg on Trem2 ***. gondii significantly suppressed Trem2 expression in both mouse placentas and cellular models, with truncated promoter assays identifying key regulatory regions of the Trem2 promoter inhibited by sTgAg. ATF3 was identified as a transcription factor responsive to sTgAg stimulation, which bound to the Trem2 promoter. Importantly, knockdown of ATF3 restored Trem2 expression, demonstrating its critical role in mediating the inhibitory effect of *** identified that sTgAg may target and inhibit Trem2 expression through the transcription factor ATF3, and inhibition of ATF3 activity may help maintain Trem2 expression in macrophages, providing a pot

关键词： Activating transcription factor 3   Sishen pill   Ulcerative colitis   Neutrophil extracellular traps   ERK/MMP9   CXCL1/CXCR2  

摘要： Background: Ulcerative colitis (UC) is a chronic inflammatory bowel disease with limited therapeutic options, particularly for moderate-to-severe cases. The present study evaluated the therapeutic potential of Sishen Pill (SSP) through Activating Transcription Factor 3 (ATF3). Methods: Colonic biopsy samples were collected from 11 UC patients and 6 healthy controls (HCs). A murine colitis model was established using dextran sodium sulfate (DSS) and treated with SSP. The therapeutic efficacy of SSP was evaluated by measuring body weight, colonic length, and inflammatory markers in wild-type (WT) mice. Transcriptomic analysis revealed differentially expressed genes in colonic tissues following Atf3 knockout. Western blotting, immunofluorescence, immunohistochemical, and Luminex assays were conducted to assess the effect of SSP on Neutrophil Extracellular Traps (NETs) formation and ATF3 signaling. Results: ATF3 expression was significantly reduced in the inflamed mucosa of UC patients, correlating with disease severity. UC tissues also exhibited increased spontaneous NETs formation. In DSS-induced colitis mice, ATF3 expression was similarly reduced, whereas SSP treatment upregulated ATF3, mitigated weight loss, reduced colonic shortening, alleviated histopathological damage, and lowered inflammatory cytokine levels. Atf3 knockout mice (Atf3-/-) displayed more severe DSS-induced colitis with enhanced immune response as compared to control littermates (Atf3+/+). Transcriptomic analyses revealed that SSP downregulated key genes involved in NETs formation pathways, tumor necrosis factor (TNF) and cytokine-cytokine receptor signaling. Experimental validation confirmed that SSP reduced the levels of NETs-related proteins [Myeloperoxidase (MPO), Peptidylarginine Deiminase 4 (PAD4), Lymphocyte Antigen 6 Complex, G (Ly6G), Neutrophil Elastase (NE), Citrullinated Histone H3 (CitH3)] in the colorectal tissue of colitis mice. It also down-regulated TNF pathway-related proteins [Ph

关键词： apoptosis   dopamine D1 receptors   glucocorticoid-associated ONFH   osteogenesis  

摘要： The nervous system plays a pivotal regulatory role in the maintenance of bone homeostasis, and the protective effects of dopamine and its receptors on bone metabolism are emerging. Despite these protective roles, the functional contribution of dopaminergic signaling, particularly through specific receptor subtypes, remains unexplored in glucocorticoid (GC)-associated osteonecrosis of the femoral head (ONFH) pathogenesis. Here, the dopamine D1 receptor (DRD1), a G protein-coupled receptor with few identified bone-related functions, is identified as a positive regulator of GC-induced apoptosis. The dopamine levels in the serum of GC-associated ONFH patients are significantly lower than those in the normal population. The protein and gene expression levels of DRD1 and the number of DRD1-positive cells are abnormally elevated in the pathological state of GC-associated ONFH, and DRD1 is expressed in osteoblasts. Overexpression of DRD1 attenuates GC-induced osteogenic inhibition and apoptosis in vivo and in vitro. Mechanistically, overexpression of DRD1 elevates cAMP levels, activates downstream protein kinase A, and inhibits GC-induced endoplasmic reticulum stress and apoptosis through the ATF3/CHOP signaling pathway, thus improving bone homeostasis. Importantly, Madopar, an FDA-approved dopaminergic agent, inhibits GC-induced osteoblastic apoptosis and ONFH via DRD1. Collectively, this study not only deciphers a previously unrecognized DRD1-mediated neuro-osteogenic axis but also repurposes an FDA-approved drug (Madopar) for precision ONFH management.

关键词： 加味香砂六君子汤   载脂蛋白A-Ⅰ   炎症   转录因子3  

摘要： 探讨加味香砂六君子汤通过ApoA-I调节ATF3影响肝脏炎症反应的作用机制。16只ApoA-I－/－小鼠，24只C57BL/6J小鼠，适应性喂养1周后，将16只ApoA-I－/－小鼠随机分为2组：高脂组、中药组；将24只C57BL/6J小鼠随机分为3组：空白对照组、高脂对照组、中药对照组。空白对照组普通饲料喂养，其余组高脂饲料喂养，8周后中药组及中药对照组给予加味香砂六君子汤灌胃。采用全自动生化分析仪检测小鼠血清总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）、谷丙转氨酶（ALT）和谷草转氨酶（AST）水平，苏木精-伊红（HE）及油红O染色观察小鼠肝组织病理形态学变化，酶联免疫吸附测定（ELISA）检测各组小鼠肝组织白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）水平，实时荧光定量PCR（RT-qPCR）及蛋白免疫印迹法（Western blot）检测各组小鼠肝组织载脂蛋白A-Ⅰ（ApoA-I）、激活转录因子3（ATF3）、Toll样受体4（TLR4）、核因子κB（NF-κB）mRNA及蛋白表达水平。结果显示，与空白对照组相比，高脂对照组小鼠血清TC、TG、HDL-C、LDL-C、ALT及AST水平显著升高（P<0.01）；血清及肝组织IL-1β、IL-6、TNF-α含量及mRNA表达显著升高（P<0.01）；肝组织ApoA-I、ATF3 mRNA及蛋白表达显著降低（P<0.01），TLR4、NF-κB mRNA及蛋白表达显著升高（P<0.01）。与高脂对照组相比，中药对照组小鼠血清TC、TG、HDL-C、LDL-C、ALT及AST水平显著降低（P<0.05，P<0.01）；血清及肝组织IL-1β、IL-6、TNF-α含量及mRNA表达显著降低（P<0.05，P<0.01）；肝组织ApoA-I、ATF3 mRNA及蛋白表达升高（P<0.05，P<0.01），TLR4、NF-κB mRNA及蛋白表达显著降低（P<0.05，P<0.01）。与高脂对照组相比，高脂组小鼠血清TC、TG、LDL-C、ALT及AST水平显著升高（P<0.01），HDL-C水平显著降低（P<0.01）；血清及肝组织IL-1β、IL-6、TNF-α含量及mRNA表达显著升高（P<0.05，P<0.01）；肝组织ATF3 mRNA及蛋白表达显著降低（P<0.05，P<0.01），TLR4、NF-κB mRNA及蛋白表达显著升高（P<0.05，P<0.01）。与中药对照组相比，中药组小鼠血清TC、TG、LDL-C、ALT及AST水平显著升高（P<0.05，P<0.01），HDL-C水平显著降低（P<0.01）；血清及肝组织IL-1β、IL-6、TNF-α含量及mRNA表达显著升高（P<0.05，P<0.01）；肝组织中ATF3 mRNA及蛋白表达显著降低（P<0.01），TLR4、NF-κB mRNA及蛋白表达显著升高（P<0.05，P<0.01）。结果表明，加味香砂六君子汤可能通过上调ApoA-I表达，激活ATF3进而抑制其下游TLR4/NF-κB通路，改善血脂异常所致炎症反应。

摘要： Head and neck squamous cell carcinoma (HNSCC) remains a prevalent and challenging cancer to treat due to its genetic heterogeneity. Cisplatin resistance is one of important causes in treatment failure of locally advanced HNSCC. ONC201, a selective dopamine receptor D2 antagonist and mitochondrial ClpP agonist, has emerged as a potential antitumor agent in various malignancies. This study explores the therapeutic potential of ONC201, alone and in combination with cisplatin, in both cisplatin-sensitive and -resistant HNSCC cells, with an emphasis on endoplasmic reticulum (ER) stress-mediated apoptosis. A cisplatin-resistant HNSCC subline (OC2-CR1) was developed via long-term drug exposure. The treatment effectiveness of ONC201 alone and cisplatin in combination on cell viability, DNA damage, reactive oxygen species (ROS) production, and stress response markers were evaluated. ONC201 exhibited potent cytotoxicity in both cisplatin-sensitive and -resistant HNSCC cells, retaining efficacy in OC2-CR1 cells. Combined treatment with ONC201 and cisplatin demonstrated synergistic inhibition of proliferation and migration, with enhanced induction of apoptosis. Mechanistically, ONC201 induced ER stress-mediated cell death via ATF4/CHOP signaling in cisplatin-sensitive cells, while ATF3/CHOP predominated in resistant cells. In vivo, combination therapy significantly suppressed tumor growth in xenograft models, including cisplatin-resistant tumors, without inducing toxicity. Immunohistochemical analysis confirmed activation of CHOP in tumor tissues. Furthermore, clinical correlation revealed that low CHOP expression in OSCC patients was associated with increased recurrence risk and inferior recurrence-free survival significantly. This study provides compelling evidence that ONC201 enhances cisplatin efficacy through distinct, stress-mediated apoptotic pathways in HNSCC. The ability of ONC201 to overcome cisplatin resistance and its synergistic antitumor effects highlight its prom

关键词： 卵巢癌   绞股蓝皂苷L   mature-tRNA-Asp-GTC   pre-tRNA-Arg-TCT   铁死亡  

摘要： 目的:探讨mature-t RNA-Asp-GTC与pre-t RNA-Arg-TCT在卵巢癌(OC)细胞铁死亡表型中作用及绞股蓝皂苷L(Gyp-L)对OC细胞mature-t RNA-Asp-GTC与pre-t RNA-Arg-TCT的调控机制。方法:采用细胞增殖与活性检测法(CCK-8)检测人卵巢腺癌细胞(OVCAR3)增殖情况,计算顺铂(5、10、15、20、25、30μmol·L^(-1))、Gyp-L(25、50、75、100、125μmol·L^(-1))及在Gyp-L作用下顺铂的半数抑制浓度(IC50)以确定后续实验的干预浓度。细胞克隆实验与划痕实验反映OVCAR3细胞的增殖与迁移能力。PANDORA-seq小RNA测序检测Gyp-L干预后细胞差异表达的转运RNA衍生的小RNA(ts RNA),通过RNAhybrid软件查找ts RNA相应的靶基因。比色法或酶联免疫吸附测定法(ELISA)检测丙二醛(MDA)、谷胱甘肽(GSH)、脂质过氧化物(LPO)水平;Ferro Orange荧光探针检测Fe2+含量;DCFH-DA荧光探针检测活性氧(ROS)含量来共同反映OVCAR3细胞铁死亡的发生。将OVCAR3细胞分为空白组、50μmol·L^(-1)Gyp-L组、100μmol·L^(-1)Gyp-L组。实时荧光定量聚合酶链式反应(Real-timePCR)检测mature-t RNA-Asp-GTC、mature-t RNA-Leu-CAA、mature-mt_t RNA-Tyr-GTA_5_end、mature-t RNAVal-CAC、mature-mt_t RNA-Glu-TTC、pre-t RNA-Arg-TCT、mature-t RNA-Asn-GTT、羟甲基双烷合酶(HMBS)、Wnt、β-连环蛋白(β-catenin)、谷胱甘肽过氧化物酶4(GPX4)、Kelch样ECH关联蛋白1(KEAP1)、核转录因子红系2相关因子2(Nrf2)、活化转录因子3(ATF3)、胱氨酸/谷氨酸逆向转运蛋白(x CT)、溶血磷脂酰胆碱酰基转移酶(LPCAT3)、花生四烯酸15-脂氧合酶(ALOX15)基因表达;蛋白免疫印迹法(Western blot)检测HMBS、Wnt、β-catenin、GPX4、KEAP1、Nrf2、ATF3、x CT、LPCAT3、ALOX15蛋白表达。结果:50μmol·L^(-1)Gyp-L组、100μmol·L^(-1)Gyp-L组、顺铂组、50μmol·L^(-1)Gyp-L+顺铂组及100μmol·L^(-1)Gyp-L+顺铂组显著抑制OVCAR3细胞增殖与迁移(P<0.05)并加剧细胞铁死亡,发生体现为ROS、MDA、LPO、Fe2+含量增多及GSH含量明显降低(P<0.05)。与空白组比较,Gyp-L有效干扰OVCAR3细胞25种ts RNA的表达(P<0.05,|log2Fc|>1);Gyp-L干预后,pre-t RNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4、pre-t RNA-Arg-TCT/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/ATF3/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/LPCAT3/ALOX15轴表达均发生明显异常(P<0.05)。结论:pre-t RNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4、pre-t RNA-Arg-TCT/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/ATF3/KEAP1/Nrf2/x CT、maturet RNA-Asp-GTC/LPCAT3/ALOX15信号通路参与OC发生发展。Gyp-L主要通过mature-t RNA-Asp-GTC/ATF3/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/LPCAT3/ALOX15信号轴激活OVCAR3细胞铁死亡发生,从而抑制OC发生发展。

关键词： ATF3   TNFR1   ANGPTL4   糖尿病肾病   肾功能   预后  

摘要： 目的:分析血清转录激活因子3(ATF3)、肿瘤坏死因子受体1(TNFR1)、血管生成素样蛋白4(ANGPTL4)水平与糖尿病肾病患者肾功能及预后的相关性。方法:选择2022年4月-2024年1月在焦作市第二人民医院就诊的152例糖尿病肾病患者为研究组,另选取同期就诊的152例单纯糖尿病患者为对照组。对比2组入院时血清ATF3、TNFR1、ANGPTL4水平及肾功能指标血钙、血磷、血肌酐及肾小球滤过率(eGFR)水平,分析其相关性;对比不同预后患者入院时血清ATF3、TNFR1、ANGPTL4水平,偏回归分析其与预后的关联性及对预后的预测价值。结果:与对照组相比,研究组入院时ATF3、TNFR1、ANGPTL4水平显著升高(P<0.05);研究组血钙、血肌酐水平均高于对照组,血磷及eGFR水平均低于对照组(P<0.05);血清ATF3、ANGPTL4、TNFR1与血磷、eGFR呈负相关,与血钙、血肌酐水平呈正相关(P<0.05);研究组预后不良患者入院时ATF3、ANGPTL4、TNFR1均高于预后良好患者(P<0.05);Logistic回归分析显示,血清ATF3、ANGPTL4、TNFR1为患者预后不良的危险因素(P<0.05);入院时血清ATF3、TNFR1、ANGPTL4水平联合预测患者预后不良的效能为0.807,均高于各指标单独预测(P<0.05)。结论:糖尿病肾病患者血清ATF3、TNFR1、ANGPTL4水平升高与患者的肾功能相关,临床检测其水平有助于预测患者的预后情况,为临床诊疗提供有效参考。

关键词： 动脉粥样硬化   转录激活因子3   炎症反应   NF-κB   冠心病猝死  

摘要： 目的 探讨转录激活因子3(ATF3)在动脉粥样硬化斑块内的表达及其调控炎症反应参与动脉粥样硬化(AS)进程的机制。方法 收集尸检案例中的人冠状动脉标本,免疫荧光和Western blotting检测ATF3蛋白表达及分布;高脂饮食12周后的载脂蛋白E基因敲除(ApoE-/-)小鼠尾静脉注射9型腺相关病毒(AAV9)敲低ATF3的表达,继续高脂喂养5周构建ATF3基因敲除的动脉粥样硬化ApoE-/-小鼠模型。麻醉处死后观察主动脉斑块结构改变,检测斑块内ATF3、炎症相关因子及NF-κB信号通路蛋白表达改变,并在体外构建ATF3的过表达质粒及siRNA干预THP-1诱导的泡沫细胞模型验证ATF3与NF-κB信号通路间关系。结果 在人冠状动脉粥样硬化斑块内,ATF3的表达增高(P<0.05),且与CD68呈部分共表达;在ATF3基因敲除后,小鼠的主动脉斑块体积增大(P<0.05),斑块内炎症相关因子(CD45、CD68、IL-1β、TNF-α)表达增强(P<0.05),NF-κB信号通路相关蛋白(PIKKα/β、P-NF-κB p65)表达增高(P<0.05),VCAM1、MMP9及MMP2表达增强(P<0.05);在离体THP-1细胞实验中验证了沉默ATF3后NF-κB信号通路进一步激活,而过表达ATF3后NF-κB信号受到抑制。结论 动脉粥样硬化诱导ATF3的表达,ATF3的缺乏会促进AS的发生,增强斑块内炎症反应,其机制可能是通过进一步激活NF-κB信号通路导致,ATF3可能是AS潜在的治疗靶点。