关键词： Bioinformatics   Biomarkers   Ferroptosis   Gout   Machine learning  

摘要： Objective: Gout is a prevalent form of chronic inflammatory arthritis, and its etiology remains incompletely understood. Ferroptosis is a form of cell death that relies on iron. As of now, the relationship between ferroptosis and gout is not entirely clear. Hence, the primary objective of this study is to employ bioinformatics methods for the analysis and identification of potential genes associated with ferroptosis in the context of gout. Methods: Utilizing both bioinformatics analysis and machine learning algorithms to systematically identify biomarkers for gout. The gout-related dataset (GSE160170) was acquired from the Gene Expression Omnibus (GEO) database. Ferroptosis-related genes were extracted from the FerrDb database. subsequently, we identified DEGs associated with ferroptosis in the context of gout. Following that, we performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses on the DEGs. Subsequently, SVM-RFE analysis and the LASSO regression model were employed for biomarker screening. Additionally, CIBERSORT software was utilized to assess the composition of twenty-two immune cells in gout, and correlation analyses between hub genes and immune cells were conducted. Results: This study screened a total of twenty-five DEGs related to Ferroptosis in healthy population and gout patient. The KEGG analysis indicates that these DEGs are predominantly enriched in: the AGE-RAGE signaling pathway, nod like receptor signaling pathway, MAPK signaling pathway, IL-17 signaling pathway, etc. The intersection of the top 10 genes identified through PPI network, SVM-RFE analysis, and LASSO regression model resulted in two hub genes, namely JUN and ATF3. Analysis of immunocyte infiltration revealed that JUN exhibited associations with various immune cells, including NK cells resting, Monocytes, Mast cells resting, etc. ATF3, on the other hand, showed associations with immune cells Mast cells resting and Eosinophels. Concl

关键词： LINC00894   ATF3   Proliferation   Apoptosis  

摘要： LINC00894 may be associated with synaptic function, but its biology function in neural cells is still unknown. In this study, LINC00894 knockdown decreased the EdU incorporated into newly synthesized DNA and cell viability in MTT or CCK-8 assay in HEK-293T and BE(2)-M17 (M17) neuroblastoma cells. And LINC00894 knockdown increased cellular apoptosis in Annexin V-FITC staining, the expression of activated Caspase3 and the level of reactive oxygen species (ROS) both in HEK-293T and M17 cells. Moreover, LINC00894 also protected cells from hydrogen peroxide induced apoptosis in in vitro models. Utilizing RNA sequencing (RNA-seq) integrated with quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunoblot, we identified that LINC00894 affected activating transcription factor 3 (ATF3) expression in HEK-293T, M17, and SH-SY5Y neuroblastoma cells. Finally, we found that ectopic expression of ATF3 restored cell proliferation and inhibited cell apoptosis in LINC00894 downregulated M17 cells. While knockdown of ATF3 also significantly increased the cell viability inhibition and apoptosis promotion induced by LINC00894 knockdown in M17 cells. Our results from in vitro models revealed that LINC00894 could promote neuronal cell proliferation and inhibit cellular apoptosis by affecting ATF3 expression.

关键词： PM2.5   Apoptosis   Inflammation   ATF3   Ovarian function  

摘要： Particulate matter 2.5 (PM2.5) pollution has emerged as a major global public health concern because of its adverse effects on human health. Our group has previously demonstrated that PM2.5 exposure can seriously impair ovarian function. However, the underlying mechanisms remain a mystery. This study verifies ovarian damage in mice, evidenced by inflammatory cell infiltration and follicular atresia, following 5 months of PM2.5 exposure via tracheal drip (35 mu g/m(3) for low dose and 150 mu g/m(3) for high dose). In addition, PM2.5 exposure inhibited the cell viability of human granulosa cells (KGN) and induced apoptosis at the concentrations of 50, 100, and 150 mu g/mL for 24 h. The apoptosis of KGN cells induced by inflammation contributes to follicular atresia. Furthermore, we conducted RNA-sequencing analysis to identify the genes and pathways triggered by PM2.5 (100 mu g/mL) exposure, which decreases the KGN cell viability. We found a significant increase in Activating Transcription Factor 3 (ATF3). Further mechanistic studies reveal a strong association between PM2.5-induced apoptosis, inflammation, and ATF3 with its downstream oxidative stress signals. In summary, the ATF3 pathway serves a vital role in the ovarian injury caused by PM2.5 exposures.

关键词： miR-378-5p   ATF3   前体脂肪细胞   增殖   分化  

摘要： 肌内脂肪含量是决定肉质的关键要素,受多种因素影响,其中重要的一项成分为遗传因素。ATF3(活化转录因子3)基因被证明可以抑制肌内前体脂肪细胞的分化,但其在绵羊方面的作用尚未被探索。基于实验室前期对2月龄和12月龄绵羊背最长肌的高通量测序结果,筛选出差异表达的miR-378-5p,通过预测发现miR-378-5p与ATF3可能具有靶向关系。本研究探讨了miR-378-5p靶向ATF3基因对绵羊前体脂肪细胞增殖和分化的影响。采用双荧光素酶报告试验验证了miR-378-5p和ATF3之间的靶向关系。过表达和干扰miR-378-5p后,通过分析实时荧光定量PCR和蛋白质免疫印迹结果显示,miR-378-5p通过靶向ATF3负向调控前体脂肪细胞的增殖,正向调控前体脂肪细胞的分化。EdU细胞增殖检测结果显示,miR-378-5p通过靶向ATF3抑制前体脂肪细胞增殖。油红O染色结果显示,miR-378-5p通过靶向ATF3促进前体脂肪细胞分化。本研究结果证实了miR-378-5p靶向ATF3抑制前脂肪细胞的增殖并促进其分化。这一发现提高了目前对绵羊脂肪沉积的理解,为进一步探究绵羊分子育种提供了理论基础。

关键词： Cadmium toxicity   Ferroptosis   Oxidative stress   Testicular injury  

摘要： Cadmium (Cd) has garnered significant attention due to reproductive toxicity in inducing ferroptosis. However, the specific mechanisms underlying Cd-induced germ cell ferroptosis remain poorly understood. This study aimed to systematically explore the molecular mechanisms of germ cell ferroptosis by investigating differential changes in transcription factors and proteins in male mice treated orally with CdCl2 (0.5 g/L) reaching postnatal day 60, alongside Leydig cell (TM3) and Sertoli cell (TM4) lines. Results demonstrated that Cd exposure led to increased iron overload and oxidative stress in mouse testes, disrupted intracellular mitochondrial morphology characteristic of ferroptosis. RNA sequencing revealed significant upregulation of Atf3 and Hmox1 in Cd-exposed germ cells, along with increased expression of ATF3 and HO-1. Intervention in ferroptosis or HO-1 effectively rescued cells from Cd-induced mortality by breaking the detrimental cycle between lipid peroxidation and HO-1 activation. Further findings showed that NRF2 and HO-1 expression was notably elevated upon ATF3 overexpression in TM3 and TM4 cells, activating the Keap1-Nrf2 pathway and triggering ferroptosis in testes, whereas NRF2 and HO-1 expression levels were reversed when ATF3 was silenced. This study provides novel insights into ATF3-mediated NRF2/HO-1 signaling in Cd-induced mitochondrial ferroptosis in testes, shedding light on the mechanisms underlying Cd-induced ferroptosis and testicular injury.

摘要： It is well known that appropriate aerobic exercise can effectively alleviate fatty liver and enhance brain function. The concept of multi-organ crosstalk coordinating disease progression has become the current research hot topic. However， there remains an urgent need to elucidate its specific mechanisms. This study aimed to explore the impact of a high-fat diet （HFD） on liver health and cognitive function， and to further uncover the regulatory effect of aerobic exercise by liver-specific ATF3 knockout (ATF3cKO) mice in a “liver-brain” axis mode. The 5-week-old C57BL/6 and ATF3cKOmice were fed with HFD for 32 weeks， and sequentially subjected to aerobic exercise intervention at the 20thweek for another 12 consecutive weeks. Meanwhile， C57BL/6 mice were provided with a normal diet as the control group. The functional parameters of liver and brain of all mice were assessed. Cognitive capacity of all mice was assessed by the Morris water maze （MWM）. Inflammatory factors in the serum and brain of mice were quantified using enzyme-linked immunosorbent assay （ELISA）， and the expression of inflammasomes was detected by immunohistochemistry （IHC）. Additionally， the activation of NF-κB and PI3K signal pathways was analyzed by Western blotting. In this study， HFD impaired hepatic and brain functions， while aerobic exercise and liver-specific ATF3 knockout suppressed inflammatory factors in the peripheral circulation through hepatoprotective mechanisms， thereby attenuating cerebral inflammation and preserving neurological integrity， as well as mitigating HFD-induced cognitive decline.

摘要： The crosstalk between adipose tissue and the liver is finely controlled to maintain metabolic health. Yet, how adipose tissue controls toxic free fatty acid overflow into the liver remains incompletely understood. Here, we show that adipocyte activating transcription factor 3 (ATF3) was induced in human or mouse obesity. Adipocyte Atf3-/- (Atf3Adi-/-) mice developed obesity, glucose intolerance, and metabolic dysfunction-associated steatohepatitis (MASH) in chow diet, high-fat diet, or Western diet-fed mice. Blocking fatty acid flux by inhibiting hepatocyte CD36, but not the restoration of hepatic AMPK signaling, prevented the aggravation of MASH in Atf3Adi-/- mice. Further studies show that the loss of adipocyte ATF3 increased lipolysis via inducing adipose triglyceride lipase, which in turn induced lipogenesis and inflammation in hepatocytes. Moreover, Atf3Adi-/- mice had reduced energy expenditure and increased adipose lipogenesis and inflammation. Our data demonstrate that adipocyte ATF3 is a gatekeeper in counteracting MASH development under physiological and pathological conditions. Using adipocyte-specific adipocyte activating transcription factor 3 (ATF3) knockout mice, this study demonstrates the crucial role of adipocyte ATF3 in preventing metabolic dysfunction-associated steatohepatitis (MASH)

关键词： Noise-induced hearing loss   Calcitriol   Oxidative stress   DNA repair   ATF3   DUSP1  

摘要： Background: Calcitriol (Cal) is the most active metabolite of vitamin D and has antioxidant and anti-inflammatory properties. The aim of this study was to investigate the role of Cal in noise-induced hearing loss (NIHL) to further elucidate the mechanism of noise-induced oxidative stress in the mouse cochlea. Methods: C57BL/6 J mice were given six intraperitoneal injections of Cal (500 ng/kg/d). After 14 days of noise exposure, auditory brainstem response (ABR) thresholds, and the cochlear outer hair cell loss rate were analysed to evaluate auditory function. Real-time fluorescence quantitative PCR, immunofluorescence and western blotting were performed in vitro after the treatment of cochlear explants with 100 mu M tert-butyl hydroperoxide (TBHP) for 2.5 h and HEI-OC1 cells with 250 mu M TBHP for 1.5 h. Results: In vivo experiments confirmed that Cal pretreatment mitigated NIHL and outer hair cell death. The in vitro results demonstrated that Cal significantly reduced TBHP-induced cochlear auditory nerve fibre degradation and spiral ganglion neuron damage. Moreover, treatment with Cal inhibited the expression of oxidative stress-related factors (3-NT and 4-HNE) and DNA damage-related factors (gamma-H2A.X) gamma- H2A.X) and attenuated TBHPinduced apoptosis in cochlear explants and HEI-OC1 cells. A total of 1479 upregulated genes and 1443 down- regulated genes were screened in cochlear tissue 1 h after noise exposure. The level of transcription factor 3 (ATF3) was significantly elevated in HEI-OC1 cells after TBHP stimulation. Gene Transcription Regulation Database (GTRD)and Cistrome database analyses revealed that the downstream target gene of ATF3 is dual specificity phosphatase 1 (DUSP1). Cistrome DB Toolkit database results showed that the transcription factor of DUSP1 was ATF3. In addition, the ChIP-PCR results indicated that ATF3 might be a direct transcription factor of DUSP1. Conclusion: The results of our study suggest that Cal attenuates NIHL and inhibits n

关键词： Skin wound healing   Activating transcription factor 3   Fibroblast   Collagen   TGF-beta receptor II  

摘要： Skin wound is an emerging health challenge on account of the high-frequency trauma, surgery and chronic refractory ulcer. Further study on the disease biology will help to develop new effective approaches for wound healing. Here, we identified a wound-stress responsive gene, activating transcription factor 3 (ATF3), and then investigated its biological action and mechanism in wound healing. In the full-thickness skin wound model, ATF3 was found to promote fibroblast activation and collagen production, resulted in accelerated wound healing. Mechanically, ATF3 transcriptionally activated TGF-beta receptor II via directly binding to its specific promoter motif, followed by the enhanced TGF-beta/Smad pathway in fibroblasts. Moreover, the increased ATF3 upon skin injury was partly resulted from hypoxia stimulation with Hif-1 alpha dependent manner. Altogether, this work gives novel insights into the biology and mechanism of stress-responsive gene ATF3 in wound healing, and provides a potential therapeutic target for treatment.

摘要： Cortical bone loss is intricately associated with ageing and coincides with iron accumulation. The precise role of ferroptosis, characterized by iron overload and lipid peroxidation, in senescent osteocytes remains elusive. We found that ferroptosis was a crucial mode of osteocyte death in cortical bone during ageing. Using a single-cell transcriptome analysis, we identified activating transcription factor 3 (ATF3) as a critical driver of osteocyte ferroptosis. Elevated ATF3 expression in senescent osteocytes promotes iron uptake by upregulating transferrin receptor 1 while simultaneously inhibiting solute carrier family 7-member 11-mediated cystine import. This process leads to an iron overload and lipid peroxidation, culminating in ferroptosis. Importantly, ATF3 inhibition in aged mice effectively alleviated ferroptosis in the cortical bone and mitigated cortical bone mass loss. Taken together, our findings establish a pivotal role of ferroptosis in cortical bone loss in older adults, providing promising prevention and treatment strategies for osteoporosis and fractures.