关键词： brain arteriovenous malformation   single-cell sequencing   endothelial cells   ATF3   experiment validation   precision medicine  

摘要： Background Brain arteriovenous malformation (BAVM) is a destructive high-flow vascular abnormality that can lead to various cerebral hemodynamic disorders. The incidence of BAVM has risen significantly in recent years, yet treatment options remain limited. Endothelial cells (ECs) have been proved to be one of the key factors leading to abnormal cerebrovascular structure. Therefore, it is important to explore the pathogenesis of the disease and develop new treatment strategies. With the rapid advancement of single-cell sequencing (scRNA-seq) and the integration of multi-omics data offers a novel perspective for precision *** We first analyzed scRNA-seq data from the GEO database. We used monocle2, CytoTRACE, and slingshot to perform pseudotime trajectory analysis on ECs. CellChat was used to analyze cell-cell communication in BAVM, and pySCENIC was used to analyze related transcription factors (TFs). Finally, transfection, CCK-8, RT-qPCR, Transwell, EdU, tube formation, and other commonly used experiments were conducted to further validate the effects of key TFs on ECs *** scRNA-seq analysis showed that ECs in BAVM had significant specificity. C0 subpopulation was the key subpopulation, showing strong proliferation and differentiation ability. This study emphasized that the midkine(MK, MDK)signaling pathway was a significant signaling pathway. Heparin-binding growth factor midkine was a secreted protein with a molecular weight of 13 kDa. Studies had shown that it can promote endothelial cell proliferation and lead to angiogenesis. Then, the C0 subpopulation was also associated with a variety of TFs, among which ATF3 played a key role in the pathogenesis of BAVM. The possibility of ATF3 affecting the progression of BAVM was verified by cell *** This study employed scRNA-seq and multi-omics analysis to elucidate the pathogenesis of BAVM, uncovering the key role of ATF3 in ECs proliferation. Targeting ATF3 provided a new

关键词： 加味香砂六君子汤   载脂蛋白A   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。

关键词： activating transcription factor 3   mitochondrial homeostasis   mitogen activated protein kinase   signaling pathway   ischemic stroke  

摘要： Mitochondrial homeostasis is crucial for preventing and treatment of ischemic stroke. This study aimed to investigate the role of activating transcription factor 3 (ATF3) in ischemic stroke and mitochondrial homeostasis. ATF3 was silenced in oxygen glucose deprivation/reperfusion (OGD/R)-treated HT22 cells to evaluate its effects on cell apoptosis and mitochondrial function. The effects of silencing ATF3 on neurological injury, infarction, adenosine triphosphate (ATP), nicotinamide adenine dinucleotide (NAD+), mitofusin 1 (MFN1) and MFN2 were evaluated in stroke rats. Transcriptome sequencing and differential expression analysis were conducted to identify differential expressed genes (DEGs) associated with silencing ATF3, followed by functional enrichment analysis. The mitogen activated protein kinase (MAPK) agonist, anisomycin, was used to investigate the regulation of ATF3 in ischemic stroke and mitochondrial homeostasis via the MAPK pathway. Silencing ATF3 increased cell viability and inhibited apoptosis of OGD/R-induced cells. In stroke rats, silencing ATF3 reduced brain water content, decreased neurological injury and alleviated cerebral infarction. Notably, silencing ATF3 significantly inhibited the production of reactive oxygen species (ROS), increased the concentrations of ATP and NAD+, and upregulated the expression of MFN1 and MFN2. Next, 4,517 DGEs associated with silencing ATF3 were mainly enriched in MAPK signaling pathway. Silencing ATF3 downregulated the expression of phosphorylation-extracellular signal-regulated kinase (p-ERK)/ERK in OGD/R cells. Anisomycin notably reversed the effect of silencing ATF3 on ischemic stroke and mitochondrial homeostasis. Silencing ATF3 attenuates ischemic stroke and improves mitochondrial homeostasis via the MAPK signaling pathway, which shares a novel direction for maintaining mitochondrial homeostasis in ischemic stroke.

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： 急性心肌梗死   经皮冠状动脉介入治疗   激活转录因子3   miR-27a-3p  

摘要： 目的探讨急性心肌梗死(AMI)患者经皮冠状动脉介入治疗(PCI)术后血清miR-27a-3p和激活转录因子3(ATF3)水平与不良预后的关系。方法选取2021年1月至2022年6月收治于山东大学齐鲁医院德州医院行PCI的AMI患者112例,根据术后发生主要不良心血管事件(MACE)情况分为MACE组(n=22)和非MACE组(n=90),另选取同时期在本院体检的健康志愿者54例为对照组。采用实时荧光定量聚合酶链式反应和酶联免疫吸附试验分别检测AMI患者的血清miR-27a-3p及ATF3水平,采用Target Scan Human网站预测miR-27a-3p及ATF3的靶向关系。采用Logistic回归分析探讨AMI患者术后发生MACE的影响因素,采用Pearson相关性分析检测miR-27a-3p与ATF3的相关性,受试者工作特征曲线预测血清miR-27a-3p与ATF3水平对AMI患者术后MACE发生的预测价值。结果与对照组比较,AMI患者的血清miR-27a-3p水平显著升高,ATF3水平显著降低,差异具有统计学意义(P<0.05);与非MACE组比较,MACE组患者的血清miR-27a-3p水平、多支病变支数占比升高,ATF3水平降低,差异具有统计学意义(P<0.05);多支病变、miR-27a-3p水平为预后不良的危险因素,ATF3水平为预后不良的保护因素(P<0.05)。miR-27a-3p水平与ATF3水平呈负相关(P<0.05),且存在靶向关系。miR-27a-3p、ATF3联合预测不良预后优于miR-27a-3p水平与ATF3水平单独预测(miR-27a-3p水平:Z=2.124,P=0.033;ATF3水平:Z=1.974,P=0.048)。结论AMI患者的血清miR-27a-3p水平升高,ATF3水平降低,血清miR-27a-3p、ATF3对AMI患者PCI术后的不良预后具有一定预测价值。

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： 翼状胬肉   ATF3   Smad4   相互作用   初步探究  

摘要： 目的初步探究转录激活因子3(ATF3)与Smad家族成员4(Smad4)在翼状胬肉中的表达及其相互作用。方法通过生物信息学方法来分析NCBI数据库中的两套数据集GSE51995和GSE2513,筛选出以正常结膜作为对照时,翼状胬肉的关键差异表达基因ATF3。并通过实时荧光定量反转录聚合酶链反应(RT-qPCR)进一步检测生物信息学分析找出的差异基因。最后结合文献查找并通过基因转录调控数据库(GTRD)预测是否存在可能与目标基因结合的靶基因Smad4,RT-qPCR验证Smad4在正常结膜与翼状胬肉组织中的表达情况,并通过免疫共沉淀(Co-IP)探究ATF3与Smad4蛋白在翼状胬肉中是否存在相互作用。结果生物信息学分析结果显示,以正常结膜组织为对照,翼状胬肉组织中存在有显著性差异的低表达基因ATF3(P<0.05)。RT-qPCR实验显示,与正常结膜组织相比,ATF3在翼状胬肉组织中低表达(P<0.001)。数据库(GTRD)中检索出Smad4基因与ATF3有两个结合位点,提示可能与ATF3存在相互作用。与正常结膜相比,RT-qPCR显示翼状胬肉组织中Smad4表达下调(P<0.01),Co-IP结果显示,在使用ATF3抗体进行免疫沉淀时,翼状胬肉组织中Smad4结合量增加(P<0.05)。结论ATF3与Smad4在翼状胬肉中存在相互作用,二者在翼状胬肉组织中的低表达及其相互作用可能与翼状胬肉的发病相关。

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。

关键词： Albiflorin   Acetaminophen-induced liver injury   ATF3   Blood-biliary barrier  

摘要： Background: Acetaminophen-induced hepatotoxicity remains a clinical challenge with limited targeted therapeutic options. While recent advances have identified blood-biliary barrier disruption as a critical pathogenic mechanism, effective interventions preserving this barrier are notably lacking. Albiflorin, a key bioactive constituent of Paeonia lactiflora Pall., exhibits unique bile acid modulation along with antioxidant and antiinflammatory activities, suggesting its potential efficacy in preventing acetaminophen-induced liver injury. However, its specific role and underlying mechanisms in alleviating acetaminophen-induced hepatotoxicity remain unclear. Objective: This study's objective was to examine the pharmacological effects and primary molecular mechanisms of albiflorin in alleviating acetaminophen-induced liver injury. Methods: An acetaminophen-induced liver injury mouse model was created using a 300 mg/kg dose of acetaminophen. The hepatoprotective effects of albiflorin were assessed through histological and biochemical analyses. Blood-biliary barrier integrity was evaluated via Evans blue dye tests, immunofluorescence, and bile acid assays. Transcriptomic analysis, gene overexpression and interference techniques, and ChIP-qPCR were employed to explore the molecular mechanisms underlying the protective effects of albiflorin. Results: Albiflorin significantly reduced acetaminophen-induced liver injury, as evidenced by improved biochemical profiles and hepatocyte morphology. It also prevented increases in blood-biliary barrier permeability and bile acid levels. RNA sequencing identified 3,184 differentially expressed genes, revealing critical pathways involved in maintaining blood-biliary barrier integrity. AF reversed the acetaminophen-induced changes in the expression of genes related to the blood-biliary barrier, particularly occludin, claudin 5, ABCG5, and ABCG8. Albiflorin protected the blood-biliary barrier and mitigated acetaminophen-induced liver inju