关键词： MITOCHONDRIA   ACTIVATION   EXPRESSION   MODEL   ATF3  

摘要： Lower back pain (LBP), which is a primary cause of disability, is largely attributed to intervertebral disc degeneration (IDD). Macrophages (M phi s) in degenerated intervertebral discs (IVDs) form a chronic inflammatory microenvironment, but how M phi s are recruited to degenerative segments and transform into a proinflammatory phenotype remains unclear. We evaluated chemokine expression in degenerated nucleus pulposus cells (NPCs) to clarify the role of NPCs in the establishment of an inflammatory microenvironment in IDD and explored the mechanisms. We found that the production of C-C motif chemokine ligand 2 (CCL2) and C-C motif chemokine ligand 7 (CCL7) was significantly increased in NPCs under inflammatory conditions, and blocking CCL2/7 and their receptor, C-C chemokine receptor type 2(CCR2), inhibited the inductive effects of NPCs on M phi infiltration and proinflammatory polarization. Moreover, activation of the integrated stress response (ISR) was obvious in IDD, and ISR inhibition reduced the production of CCL2/7 in NPCs. Further investigation revealed that activating Transcription Factor 3 (ATF3) responded to ISR activation, and ChIP-qPCR verified the DNA-binding activity of ATF3 on CCL2/7 promoters. In addition, we found that Toll-like receptor 4 (TLR4) inhibition modulated ISR activation, and TLR4 regulated the accumulation of mitochondrial reactive oxygen species (mtROS) and double-stranded RNA (dsRNA). Downregulating the level of mtROS reduced the amount of dsRNA and ISR activation. Deactivating the ISR or blocking CCL2/7 release alleviated inflammation and the progression of IDD in vivo. Moreover, M phi infiltration and IDD were inhibited in CCR2-knockout mice. In conclusion, this study highlights the critical role of TLR4/mtROS/dsRNA axis-mediated ISR activation in the production of CCL2/7 and the progression of IDD, which provides promising therapeutic strategies for discogenic LBP. Low back pain, a main reason for disability, is often due to inter

关键词： Fibromyalgia   Myalgic encephalomyelitis   Intermittent cold stress   Chronic fatigue syndrome   Neuronal inflammation   Pain   Microglia   TRANSCRIPTION FACTOR-3   PITUITARY-GLAND   C-JUN   EXPRESSION   ATF3   HYPERALGESIA   MOTONEURONS   SUPPRESSION   ALLODYNIA   RATS  

摘要： BackgroundFibromyalgia is characterized by chronic pain, fatigue, and other somatic symptoms. We have recently revealed that proprioceptor hyperactivation induces chronic pain in a rat model of myalgic encephalomyelitis. The present study explores whether similar proprioceptor-induced pain is elicited in a mouse model of *** cold stress (RCS) was used as a fibromyalgia model. Pain behavior was examined using the von Frey test, and neuronal activation was examined immunohistochemically as activating transcription factor (ATF)3 expression. The Atf3:BAC transgenic mouse, in which mitochondria in hyperactivated neurons are specifically labeled by green fluorescent protein, was used to trace the activated neuronal circuit. PLX3397 (pexidartinib) was used for microglial *** elicited long-lasting pain in mice. ATF3, a marker of cellular hyperactivity and injury, was expressed in the lumbar dorsal root ganglion (DRG) 2 days after RCS initiation; the majority of ATF3-expressing DRG neurons were tropomyosin receptor kinase C- and/or vesicular glutamate transporter 1-positive proprioceptors. Microglial activation and increased numbers of microglia were observed in the medial part of the nucleus proprius 5 days after RCS initiation, and in the dorsal region of the ventral horn 7 days after RCS. In the ventral horn, only a subset of motor neurons was positive for ATF3; these neurons were surrounded by activated microglia. A retrograde tracer study revealed that ATF3-positive motor neurons projected to the intrinsic muscles of the foot (IMF). Using Atf3:BAC transgenic mice, we traced hyperactivated neuronal circuits along the reflex arc. Green fluorescent protein labeling was observed in proprioceptive DRG neurons and their processes originating from the IMF, as well as in motor neurons projecting to the IMF. Microglial activation was observed along this reflex arc, and PLX3397-induced microglial ablation significantly suppressed pain b

关键词： CHRONIC-FATIGUE-SYNDROME   MONOCYTE RECRUITMENT   DIFFERENTIATION   EXPRESSION   SEVERITY   GENES   ATF3  

摘要： Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a serious and poorly understood disease. To understand immune dysregulation in ME/CFS, we use single -cell RNA sequencing (scRNA-seq) to examine immune cells in patient and control cohorts. Postexertional malaise (PEM), an exacerbation of symptoms following strenuous exercise, is a characteristic symptom of ME/CFS. To detect changes coincident with PEM, we applied scRNA-seq on the same cohorts following exercise. At baseline, ME/CFS patients display classical monocyte dysregulation suggestive of inappropriate differentiation and migration to tissue. We identify both diseased and more normal monocytes within patients, and the fraction of diseased cells correlates with disease severity. Comparing the transcriptome at baseline and postexercise challenge, we discover patterns indicative of improper platelet activation in patients, with minimal changes elsewhere in the immune system. Taken together, these data identify immunological defects present at baseline in patients and an additional layer of dysregulation in platelets.

关键词： Epilepsy   Ferroptosis   FUS   ATF3   EXPRESSION  

摘要： Background: Epilepsy is a neurological disorder characterized by recurrent seizures and is often unresponsive to current treatment options. Ferroptosis, a recently defined iron-dependent regulated cell death, has been suggested as a potential therapeutic target for epilepsy due to its association with oxidative stress. Additionally, circRNA SLC8A1 (circSLC8A1) has been implicated in various neurological disorders and oxidative stress-related diseases but its involvement in epilepsy progression, particularly in relation to ferroptosis and oxidative stress, remains ***: qRT-PCR, Western blot, IHC and ELISA assays were employed to validate the relative expression of targeted genes and proteins. The levels of ROS, iron, LOP and GSH were detected by commercial kits. RNA pulldown and RIP assays were employed to detect the interactions among circSLC8A1, FUS and ATF3. A rat epilepsy model was established for further in vivo confirmation. Results and conclusion: In this study, we investigated the potential involvement of circSLC8A1 in epilepsy progression and its connection to ferroptosis and oxidative stress. Our findings demonstrate that circSLC8A1 triggers neuronal ferroptosis by stabilizing ATF3 mRNA expression through recruitment with FUS. The induced neuronal ferroptosis contributes to epilepsy progression. These results enhance our understanding of epilepsy pathogenesis and may provide insights for the development of novel therapeutic strategies.

关键词： Carfilzomib   ATF3   LDHA   Metabolic reprogramming   Esophageal squamous cell carcinoma   ACTIVATING TRANSCRIPTION FACTOR-3   ENDOPLASMIC-RETICULUM STRESS   LACTATE-DEHYDROGENASE   UP-REGULATION   CANCER   GENE   APOPTOSIS   MITOCHONDRIA   EXPRESSION   BORTEZOMIB  

摘要： Carfilzomib, a second-generation proteasome inhibitor, has been approved as a treatment for relapsed and/or refractory multiple myeloma. Nevertheless, the molecular mechanism by which Carfilzomib inhibits esophageal squamous cell carcinoma (ESCC) progression largely remains to be determined. In the present study, we found that Carfilzomib demonstrated potent anti-tumor activity against esophageal squamous cell carcinoma both in vitro and in vivo. Mechanistically, carfilzomib triggers mitochondrial apoptosis and reprograms cellular metabolism in ESCC cells. Moreover, it has been identified that activating transcription factor 3 (ATF3) plays a crucial cellular target role in ESCC cells treated with Carfilzomib. Overexpression of ATF3 effectively antagonized the effects of carfilzomib on ESCC cell proliferation, apoptosis, and metabolic reprogramming. Furthermore, the ATF3 protein is specifically bound to lactate dehydrogenase A (LDHA) to effectively suppress LDHA-mediated metabolic reprogramming in response to carfilzomib treatment. Research conducted in xenograft models demonstrates that ATF3 mediates the anti-tumor activity of Carfilzomib. The examination of human esophageal squamous cell carcinoma indicated that ATF3 and LDHA have the potential to function as innovative targets for therapeutic intervention in the treatment of ESCC. Our findings demonstrate the novel function of Carfilzomib in modulating ESCC metabolism and progression, highlighting the potential of Carfilzomib as a promising therapeutic agent for the treatment of ESCC.

关键词： MicroRNA-124   SAH   AK4   Neuroprotective effect   CEREBROSPINAL-FLUID   BRAIN   MIR-124   MUSCLE   EXPRESSION   APOPTOSIS   ANEURYSMS   PROTECTS   GELSOLIN   NEURONS  

摘要： Spontaneous subarachnoid hemorrhage (SAH) accounts for approximately 5% of all cases of stroke. SAH is correlated with elevated rates of mortality and disability. Despite significant advancements in comprehending the pathogenesis and surgical management, efficacious clinical interventions remain restricted, and the prognosis is yet to be enhanced. MicroRNAs play a crucial role in various pathological processes in organisms. Revealing these regulatory processes is conducive to the development of new treatment methods. MicroRNA-124 is highly expressed in the nervous system and has significant research value for SAH. This study aims to explore the role of miR-124 in the early post-SAH period on neural function and verify whether it is involved in the pathological and physiological processes of SAH. In this study, we used methods such as comparing the expression levels of miR-124 in cerebrospinal fluid, establishing a rat SAH model, and a mouse embryonic primary neuron hemoglobin stimulation model to verify the downstream proteins of miR-124 in SAH. Through transfection techniques, we adjusted the expression of this small RNA in Vitro and in Vivo models using miR-124 inhibitor and mimic in the primary neuron hemoglobin stimulation model and rat SAH model, and observed the phenotype. Finally, by consulting the literature and verifying in Vivo and in Vitro methods, AK4 and downstream molecule ATF3 were identified as downstream targets of miR-124.

摘要： Pro-reparative cardiac-resident macrophages have emerged as major players in salvaging the ischemic myocardium of a diseased heart. New research now highlights ATF3 as a key transcription factor that governs macrophage survival and proliferation and myocardial repair.

关键词： RECEPTOR TYROSINE KINASE   MYOCARDIAL-INFARCTION   CELL-SURVIVAL   HEART-FAILURE   REPAIR   PATHWAYS   DELIVERY   HYPOXIA   AXIS  

摘要： Cardiac resident MerTK+ macrophages exert multiple protective roles after ischemic injury; however, the mechanisms regulating their fate are not fully understood. In the present study, we show that the GAS6-inducible transcription factor, activating transcription factor 3 (ATF3), prevents apoptosis of MerTK+ macrophages after ischemia-reperfusion (IR) injury by repressing the transcription of multiple genes involved in type I interferon expression (Ifih1 and Ifnb1) and apoptosis (Apaf1). Mice lacking ATF3 in cardiac macrophages or myeloid cells showed excessive loss of MerTK+ cardiac macrophages, poor angiogenesis and worse heart dysfunction after IR, which were rescued by the transfer of MerTK+ cardiac macrophages. GAS6 administration improved cardiac repair in an ATF3-dependent manner. Finally, we showed a negative association of GAS6 and ATF3 expression with the risk of major adverse cardiac events in patients with ischemic heart disease. These results indicate that the GAS6-ATF3 axis has a protective role against IR injury by regulating MerTK+ cardiac macrophage survival and/or proliferation. Shao, Li, Liu et al. identify the GAS6-ATF3 axis as a central regulator of survival and proliferation of protective resident MerTK+ cardiac macrophages and show that GAS6 administration improves cardiac repair after ischemic-reperfusion injury in mice in an ATF3-dependent manner.

关键词： Activating transcription factor 3   Ferroptosis   Human periodontal stem cells   Nrf2/HO-1   Osteogenic differentiation   Periodontitis  

摘要： Activating transcription factor 3 (ATF3) has been identified as a regulator associated with osteoblast differentiation. However, the effects of ATF3 on the osteogenic differentiation and proliferation of human periodontal stem cells (hPDLSCs) in periodontitis have not been reported. With the purpose of establishing an in vitro model of periodontitis, hPDLSCs were challenged with lipopolysaccharide (LPS). The Cell Counting Kit-8 assay was applied to assess cell viability, while reverse transcription-quantitative PCR and western blotting were employed to detect ATF3 expression. Inflammatory release was assessed using ELISA, together with western blotting. Lipid peroxidation was explored using the C11 BODIPY 581/591 probe, biochemical kits, thiobarbituric acid reactive substances (TBARS) assay and DCFH-DA staining. Iron and Fe2+ levels, and the expression levels of ferroptosis-related proteins were measured using corresponding kits and western blotting. Osteogenic differentiative capability was evaluated using alkaline phosphatase staining, Alizarin red staining and western blotting. The expression levels of proteins associated with Nrf2/HO-1 signaling were identified using western blotting. The results indicated that ATF3 expression was upregulated in LPS-induced hPDLSCs. The knockdown of ATF3 alleviated the LPS-induced inflammatory response in hPDLSCs, together with increased levels of TNF-α, IL-6, IL-1β, Cox-2 and iNOS, and decreased levels of IL-10. ATF3 silencing also led to lower TBARS production rate, and reduced levels of reactive oxygen species, iron, Fe2+, ACSL4 and TFR1, whereas it elevated the levels of SLC7A11 and GPX4. In addition, ATF3 silencing promoted hPDLSC mineralization and cell differentiation, and elevated the levels of OCN2, RUNX2 and BMP2. Additionally, ATF3 depletion upregulated the expression levels of proteins related with Nrf2/HO-1 signaling. The Nrf2 inhibitor ML385 partially counteracted the effects of ATF3 interference on the LPS-challen

关键词： 七氟烷   铁死亡   内质网应激   ATF3   过氧化氢   神经发育毒性  

摘要： 研究背景:随着麻醉和外科手术技术的快速进步和发展,越来越多的婴幼儿及孕产妇接受麻醉和手术治疗。人类的神经系统在胎儿妊娠末期至婴幼儿出生后2～3年内是发育的重要时期,发育期大脑极易受到包括全麻药物在内的各种因素的干扰和影响,可能产生发育脑神经毒性损伤。大量动物研究和临床数据表明,在生命发育的早期阶段暴露吸入麻醉药七氟烷、异氟烷能够引起婴幼个体未成熟大脑产生广泛的神经病理性改变,导致长期的行为学异常和神经认知功能损害。因此,探讨吸入麻醉药的神经发育毒性作用及其机制成为麻醉学乃至神经科学急需解决的问题。研究表明导致神经元死亡是吸入麻醉药诱导发育脑神经毒性的主要特征,但具体机制不清。铁死亡（ferroptosis）是细胞程序性死亡的一种形式,其特征是细胞内亚铁离子(Fe)增加,积聚的Fe能够与过氧化氢（HO）通过芬顿反应生成羟基自由基,进而对多不饱和脂肪酸（PUFA）产生脂质过氧化损伤。研究证实,铁死亡不仅介导心脏、肝脏和肾脏等器官损伤的细胞死亡过程,还与多种神经疾病的细胞死亡相关。内质网不仅是细胞内蛋白质合成、修饰、折叠加工的场所,也是驱动脂质过氧化的中枢枢纽。细胞感受内外环境的伤害刺激后,可引起内质网应激,其中PERK/ATF4通路活化与铁死亡的发生密切相关。作为一种应激反应因子,激活转录因子3（ATF3）在细胞受到多种伤害性刺激时迅速表达,进而调控相关靶基因转录激活或抑制。在内质网应激情况下,PERK/ATF4通路活化能够进一步激活ATF3,在多种形式的细胞死亡,包括细胞凋亡、自噬、铁死亡中起重要作用。目前,七氟烷能否诱导发育脑神经元铁死亡仍未明确。内质网应激介导的ATF3活化在七氟烷诱导的发育脑神经元铁死亡过程中作用如何,其调控细胞内铁离子稳态失衡的机制是什么,以及脂质过氧化损伤级联通过什么途径被激活尚不清楚。研究目的:（1）探究七氟烷能否诱导发育脑神经元铁死亡。（2）探究内质网应激介导的ATF3活化在七氟烷诱导发育脑铁死亡过程中可能的作用机制,为吸入麻醉药的神经发育毒性作用提供一种潜在的防治策略。研究方法:（1）HT22小鼠海马神经元细胞和大鼠原代海马神经元分别给予2%、4%、8%七氟烷处理6 h、12 h、24 h后,采用MTT法检测细胞的活力,LDH释放实验检测细胞的死亡率,应用铁离子和丙二醛（MDA）检测试剂盒检测神经元细胞内Fe浓度和MDA含量,应用光学显微镜观察神经元细胞的形态变化,应用透射电镜观察神经元细胞线粒体形态的变化,应用蛋白印迹法检测神经元细胞内铁离子代谢相关蛋白包括转铁蛋白受体（TFR）、转铁蛋白（TF）和铁转运蛋白（FPN）的表达。神经元细胞分别给予铁螯合剂去铁胺（DFO）(100μmol/L),抗氧化剂Fer-1（50μmol/L）、Lip-1（10μmol/L）、Vit E（100μmol/L）、GSH（2.5 mmol/L）或等量生理盐水预处理1 h,然后分别给予4%、8%七氟烷处理24 h后,检测细胞的生存率和死亡率及神经元细胞内Fe浓度和MDA含量。（2）HT22小鼠海马神经元细胞和大鼠原代海马神经元分别给予4%、8%七氟烷处理6～24 h,应用HO试剂盒检测神经元细胞内HO含量。神经元细胞分别给予GSH（2.5 mmol/L）或生理盐水预处理1 h,然后给予七氟烷处理,应用铁离子和过氧化氢试剂盒检测神经元细胞内Fe浓度和HO含量,应用蛋白印迹法检测神经元细胞内TFR、TF和FPN的表达。神经元细胞给予HO（500μmol/L）处理24 h后,检测神经元细胞内死亡率、神经元细胞内Fe浓度和MDA含量及TFR、TF和FPN的表达。（3）HT22小鼠海马神经元细胞和大鼠原代海马神经元给予4%、8%七氟烷处理6～24 h后,应用蛋白印迹法检测神经元细胞内ATF3的表达,应用共聚焦显微镜检测神经元细胞核内ATF3的聚集。神经元细胞应用si RNA沉默ATF3基因,再给予8%七氟烷处理24 h后,检测细胞的死亡率,应用铁离子、过氧化氢和MDA试剂盒检测神经元细胞内Fe、HO和MDA含量,应用蛋白印迹法检测神经元细胞内TFR、TF和FPN的表达。（4）HT22小鼠海马神经元细胞和大鼠原代海马神经元分别给予4%、8%七氟烷处理6～24 h后,应用蛋白印迹法检测神经元细胞内质网应激相关蛋白GRP78、PERK和ATF4的表达。神经元细胞给予内质网应激抑制剂4-PBA（250μmol/L）、PERK抑制剂GSK2606414（1μmol/L）或生理盐水预处理1 h,然后给予8%七氟烷处理24 h后,检测细胞的死亡率,应用铁离子、过氧化氢和MDA试剂盒检测神经元细胞内Fe、HO和MDA含量,应用蛋白印迹法检测神经元细胞GRP78、PERK、ATF4、ATF3、TFR、TF和FPN的表达。神经元细胞应用si RN