关键词： OBSTRUCTIVE PULMONARY-DISEASE   CHRONIC MUCUS HYPERSECRETION   ATF3   JUN   FOS   MORTALITY   CELLS   COPD   AP-1  

摘要： Mucus hypersecretion is an important pathologic feature of chronic obstructive pulmonary disease. Activating transcription factor 3 (ATF3) is an adaptive-response gene that participates in various cellular processes. However, Little is known about its role in cigarette smoke (CS)-induced mucus hyperproduction. This study aimed to investigate the role and molecular mechanisms of ATF3 in CS-induced Mucin 5AC (MUC5AC) expression. ATF3 was elevated in lung tissues of mice exposed to CS for 12 weeks. Treatment with CS extract significantly induced ATF3 expression and MUC5AC production in human bronchial epithelial cells, NCI-H292, and mouse tracheal epithelial cells. Interference of ATF3 significantly attenuated CS-induced MUC5AC expression in NCI-H292 and human bronchial epithelial cells. Mouse tracheal epithelial cells isolated from Atf3(-/-) mice also exhibited less MUC5AC production in response to CS extract treatment. In vivo, the Atf3(-/-) mice also displayed a significantly reduced mucus production relative to wild-type controls in response to chronic CS exposure. Furthermore, a chromatin immunoprecipitation assay revealed increased ATF3 binding to the MUC5AC promoter after CS treatment, and this transcriptional binding was significantly inhibited by knockdown of JUN, a subunit of activator protein-1. These results demonstrate that ATF3 may be involved in activator protein-1 signaling and transcriptional promotion of CS-induced MUC5AC expression in airway epithelial cells.

关键词： Type 2 diabetes   Cardiac remodeling   Cytokines   High fat diet   ATF3   ACTIVATING TRANSCRIPTION FACTOR-3   ADAPTIVE-RESPONSE GENE   STRESS-INDUCIBLE GENE   BETA-CELL APOPTOSIS   DIABETIC CARDIOMYOPATHY   METABOLIC MECHANISMS   CARDIAC-HYPERTROPHY   TRANSGENIC MICE   ADIPOSE-TISSUE   MOUSE  

摘要： Aims Obesity and type 2 diabetes (T2D) trigger a harmful stress-induced cardiac remodeling process known as cardiomyopathy. These diseases represent a serious and widespread health problem in the Western world; however the underlying molecular basis is not clear. ATF3 is an ` immediate early' gene whose expression is highly and transiently induced in response to multiple stressors such as metabolic, oxidative, endoplasmic reticulum and inflammation, stressors that are involved in T2D cardiomyopathy. The role of ATF3 in diabetic cardiomyopathy is currently unknown. Our research has aimed to study the effect of ATF3 expression on cardiomyocytes, heart function and glucose homeostasis in an obesity-induced T2D mouse model. Methods and results We used wild type mice (WT) as well as mutant mice with a cardiac-specific ATF3 deficiency (ATF3-cKO). Mice were fed a high-fat diet (HFD) for 15 weeks. HFD induced high ATF3 expression in cardiomyocytes. Mice were examined for cardiac remodeling processes and the diabetic state was assessed. HFD-fed ATF3-cKO mice exhibited severe cardiac fibrosis, higher levels of heart hypertrophic markers, increased inflammation and worse cardiac function, as compared to WT mice. Interestingly, HFD-fed ATF3-cKO mice display increased hyperglycemia and reduced glucose tolerance, despite higher blood insulin levels, as compared to HFD-fed WT mice. Elevated levels of the cardiac inflammatory cytokines IL-6 and TNFa leading to impaired insulin signalling may partially explain the peripheral glucose intolerance. Conclusions Cardiac ATF3 has a protective role in dampening the HFD-induced cardiac remodeling processes. ATF3 exerts both local and systemic effects related to T2D-induced cardiomyopathy. This study provides a strong relationship between heart remodeling processes and blood glucose homeostasis.

关键词： inflammation   microarray   phenotype   ADAPTIVE-RESPONSE GENE   NEGATIVE REGULATOR   ATF3   MECHANISMS   BIOLOGY   STRESS   INFLAMMATION   METABOLISM   MACROPHAGE   RESISTANCE  

摘要： Activating transcription factor (ATF)3 regulates the expression of inflammation-related genes in several tissues under pathological contexts. In skeletal muscle, atf3 expression increases after exercise, but its target genes remain unknown. We aimed to identify those genes and to determine the influence of ATF3 on muscle adaptation to training. Skeletal muscles of ATF3-knockout (ATF3-KO) and control mice were analyzed at rest, after exercise, and after training. In resting muscles, there was no difference between genotypes in enzymatic activities or fiber type. After exercise, a microarray analysis in quadriceps revealed ATF3 affects genes modulating chemotaxis and chemokine/cytokine activity. Quantitative PCR showed that the mRNA levels of chemokine C-C motif ligand (ccl)8 and chemokine C-X-C motif ligand (cxcl)13 were higher in quadriceps of ATF3-KO mice than in control mice. The same was observed for ccl9 and cxcl13 in soleus. Also in soleus, ccl2, interleukin (il)6, il1, and cluster of differentiation (cd)68 mRNA levels increased after exercise only in ATF3-KO mice. Endurance training increased the basal mRNA level of hexokinase-2, hormone sensitive lipase, glutathione peroxidase-1, and myosin heavy chain IIa in quadriceps of control mice but not in ATF3-KO mice. In summary, ATF3 attenuates the expression of inflammation-related genes after exercise and thus facilitates molecular adaptation to training.

关键词： NF-KAPPA-B   DIABETIC CARDIOMYOPATHY   ATF3   CARDIOMYOCYTES   REGULATOR   MICE  

关键词： Activating transcriptional factor 3   Acute kidney injury   Biomarker   Neutrophil gelatinase-associated lipocalin   Urine exosome   GELATINASE-ASSOCIATED LIPOCALIN   CRITICALLY-ILL PATIENTS   CYSTATIN C   RENAL-DISEASE   DISCOVERY   GENE   CREATININE   ATF3   COLLECTION   GUIDELINES  

摘要： Background: An early sepsis-induced acute kidney injury (sepsis-AKI) biomarker is currently in needed. Urinary neutrophil gelatinase-associated lipocalin (uNGAL) is a candidate of sepsis-AKI biomarker but with different cut-point values. Urinary exosomal activating transcriptional factor 3 (uATF3) has been mentioned as an interesting biomarker. Methods: We conducted experiments in mice and a prospective, multicenter study in patients as a proof of concept that urine exosome is an interesting biomarker. An early expression of ATF3 in kidney of CD-1 mice at 6 h after cecal ligation and puncture implied the possibility of uATF3 as an early sepsis-AKI biomarker. Increase serum creatinine (Scr) >= 0.3 mg/dL from the baseline was used as an AKI diagnosis and urine was analyzed for uATF3 and uNGAL. Patients with baseline Scr at admission >= 1.5 mg/dL were excluded. Results: The analysis showed higher Scr, uNGAL and uATF3 in patients with sepsis-AKI in comparison with patients with sepsis-non-AKI and healthy volunteers. A fair correlation, r(2) = 0.47, between uATF3 and uNGAL was showed in sepsis-AKI group with Scr >= 2 mg/dL. To see if uATF3 could be an early sepsis-AKI biomarker, urine sample was collected daily during the first week of the admission. In sepsis-AKI and sepsis-non-AKI groups, uNGAL were 367 +/- 43 ng/mL and 183 +/- 23 ng/mL, respectively; and uATF3 were 19 +/- 4 ng/mL and 1.4 +/- 0.8 ng/mL, respectively. With the mean value of uNGAL and uATF3 in sepsis AKI as a cut-off level, AUROC of uNGAL and uATF3 were 64% (95% CI 0.54 to 0.74) and 84% (95% CI 0.77 to 0.91), respectively. Conclusions: Urine exosome is an interesting source of urine biomarker and uATF3 is an interesting sepsis-AKI biomarker.

关键词： NECROSIS-FACTOR-ALPHA   ACTIVATING TRANSCRIPTION FACTOR-3   NF-KAPPA-B   WALL SHEAR-STRESS   IN-VIVO   ADHESION MOLECULES   OXIDATIVE STRESS   DISTURBED FLOW   TOBACCO-SMOKE   E-SELECTIN  

摘要： Endothelial dysfunction caused by the combined action of disturbed flow, inflammatory mediators and oxidants derived from cigarette smoke is known to promote coronary atherosclerosis and increase the likelihood of myocardial infarctions and strokes. Conversely, laminar flow protects against endothelial dysfunction, at least in the initial phases of atherogenesis. We studied the effects of TNF alpha and cigarette smoke extract on human coronary artery endothelial cells under oscillatory, normal laminar and elevated laminar shear stress for a period of 72 hours. We found, firstly, that laminar flow fails to overcome the inflammatory effects of TNF alpha under these conditions but that cigarette smoke induces an anti-oxidant response that appears to reduce endothelial inflammation. Elevated laminar flow, TNF alpha and cigarette smoke extract synergise to induce expression of the transcriptional regulator activating transcription factor 3 (ATF3), which we show by adenovirus driven overexpression, decreases inflammatory gene expression independently of activation of nuclear factor-kappa B. Our results illustrate the importance of studying endothelial dysfunction in vitro over prolonged periods. They also identify ATF3 as an important protective factor against endothelial dysfunction. Modulation of ATF3 expression may represent a novel approach to modulate proinflammatory gene expression and open new therapeutic avenues to treat proinflammatory diseases.

关键词： MOLECULAR-MECHANISMS   SIGNALING PATHWAYS   RAW264.7 CELLS   KAPPA-B   KINASE   INVOLVEMENT   LOBULARIS   ATF3   LIPOPOLYSACCHARIDE   ACTIVATION  

摘要： A new anthranilic acid derivative (1) was isolated from a Philippine sponge, Oscarella stillans (Bergquist and Kelly). The structure of compound 1, named oscarellin, was determined as 2-amino-3-(3'-aminopropoxy)benzoic acid from spectroscopic data and confirmed by synthesis. We examined the immunomodulating effect of compound 1 and its mechanism in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Our data indicated that the expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-6 were significantly reduced by the pretreatment of 1 (0.1-10 mu M) for 2 h. In addition, compound 1 suppressed activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun NH2-termimal kinase (JNK), but not p38 mitogen-activated protein kinase (MAPK) in LPS-stimulated RAW 264.7 cells. Compound 1 abrogated LPS-induced nuclear factor-kappa B (NF-kappa B) and activator protein-1 (AP-1) activities, whereas the induction of activating transcription factor-3 (ATF-3) was increased. Taken together, our results suggest that compound 1 attenuates pro-inflammatory cytokines via the suppression of JNK, ERK, AP-1, and NF-kappa B and the activation of the ATF-3 signaling pathway.

关键词： 猪血凝性脑脊髓炎病毒   ATF2   表达与调控  

摘要： 血凝性脑脊髓炎病毒主要感染1～3周龄的哺乳仔猪,常被称为猪血凝性脑脊髓炎病毒(Porcine hemagglutinating encephalomyelitis virus,PHEV),仔猪感染后可出现明显的神经症状,病死率可达20%～100%。小鼠可感染PHEV,常作为感染PHEV的病理模型。本课题组前期研究表明小鼠感染PHEV后,激活转录因子3(Activating transcription factor 3,ATF3)的表达也增加。ATF3可作为ATF2的下游靶基因,ATF2为JNK信号通路中的一员。除JNK信号通路外,ATF3还可由P53、TGF-β/smad和c-Myc等信号通路诱导激活。但是前期研究中仅发现JNK信号通路相关因子被激活。据此推测PHEV感染小鼠后,大脑中ATF3的高表达可能是由JNK信号通路所诱导的。若此推论成立,作为调控ATF3的上游基因ATF2也应呈现高表达。为验证上述假设,本实验对感染PHEV后不同时间段的小鼠大脑中的ATF2和ATF3的表达水平进行检测。本实验首先构建了 PHEV急性感染小鼠的模型,采用颅内接种方式,以小鼠在接毒后第5天全部死亡的接种剂量最为最佳接种剂量。选取4周龄SPF雌性昆明小鼠30只,随机分两组,接毒组和对照组,每组15只。接毒组以最佳接种剂量颅内接种PHEV,对照组以相同剂量颅内接种PBS。于接毒后的第1天,2天,3天,4天,5天每组随机选取3只小鼠进行剖杀。采取大脑,一部分固定于福尔马林溶液,另一部分冻存于液氮中。固定于福尔马林溶液中的大脑用于病理组织学病变的观察;冻存于液氮中的大脑,采用Real-time PCR和Western Blot法检测ATF2和ATF3 mRNA和蛋白在大脑内的表达情况。采用2-△△Ct相对表达量检测ATF2、ATF3和PHEVmRNA的表达水平,采用半定量法检测其蛋白的表达水平,并采用SPSS 17.0软件进行数据分析和处理。结果显示,小鼠感染PHEV后主要表现为非化脓性脑炎的病理组织学变化,且随着感染时间的延长,病理损伤逐渐加重。同时,随着感染时间的延长,ATF2、ATF3与PHEV的mRNA和蛋白表达水平均呈逐渐上升趋势。经相关性分析,ATF2和ATF3的表达呈正相关(p<0.01),ATF2表达与PHEV之间呈正相关(p<0.01),ATF3和PHEV之间表达呈正相关(p<0.01)。接毒第5天达到峰值,经One-Way ANOVA分析均具有显著性差异(p<0.01)。说明小鼠感染PHEV后大脑中的病理损伤逐步加重,且随着ATF2的表达量的增高,ATF3表达和PHEV病毒载量也增高。小鼠感染PHEV后ATF3的高表达可能是JNK信号通路激活后由ATF2诱导调控的。本研究为进一步探明PHEV感染的神经细胞过程中ATF3高表达的机制指引了新方向,为深入解析PHEV的分子致病机制提供重要的理论依据。

关键词： 猪血凝性脑脊髓炎病毒   ATF3   JNK抑制剂   细胞周期与凋亡  

摘要： 猪血凝性脑脊髓炎是由猪血凝性脑脊髓炎病毒(Porcine hemagglutinating encephalomyelitis virus,PHEV)引起的,主要感染3周龄的仔猪,临床以呕吐衰竭和明显的神经症状为特征。前期研究结果表明,小鼠感染PHEV后大脑中ATF3呈高表达,为了探明ATF3在神经细胞感染PHEV后ATF3是否呈现高表达,以及其高表达的机制,本实验对神经细胞感染PHEV前后ATF3的表达进行了检测,同时探索JNK信号通路在猪血凝性脑脊髓炎病毒感染神经细胞过程中对ATF3表达的影响。本试验以N2a细胞作为研究PHEV感染神经细胞的模型,根据试验要求,设置4组:对照组、接毒组、JNK抑制剂组和JNK抑制剂接毒组,每组3个重复。于六孔板中培养N2a细胞到单层,以吸附法接种PHEV,然后分别在2h、12h、24h时提取细胞总RNA和总蛋白,通过荧光定量PCR和蛋白质印迹法(Western Blot)分别测定PHEV感染前后及JNK抑制剂作用前后N2a细胞中ATF3 mRNA和蛋白的表达量。同时设置6组:对照组、接毒组、DMSO组、DMSO接毒组、JNK抑制剂组和JNK抑制剂接毒组,每组3个重复,通过Annexin V-FITC/PI双染法和PI染色法检测12h和24h的细胞凋亡和细胞周期,以测定JNK抑制剂作用前后对PHEV感染的神经细胞的凋亡和细胞周期的影响。结果表明,ATF3基因在正常N2a细胞中的处于低表达,随着PHEV mRNA和蛋白表达量的降低或升高ATF3 mRNA和蛋白的表达量也随着降低或升高,两者之间的变化趋势一致且呈正相关;JNK抑制剂作用N2a细胞后,ATF3的mRNA和蛋白表达量明显降低。通过Annexin V-FITC/PI双染法标记凋亡细胞,再经流式细胞仪进行检测,结果发现与对照组相比,JNK抑制剂作用后接毒组细胞凋亡率明显上升,差异极显著(P<0.01),此时ATF3表达量降低,说明ATF3在细胞内起到抑制凋亡的作用;同时,与对照组相比,JNK抑制剂作用后接毒组细胞周期发生阻滞。N2a细胞感染PHEV后,ATF3的表达量增高。JNK抑制剂作用于N2a细胞后,ATF3表达量下降,细胞凋亡率上升,细胞周期阻滞,DNA合成受阻,细胞活力下降,抗病毒能力下降。表明神经细胞感染PHEV后JNK信号通路可调控ATF3的表达,进而影响神经细胞凋亡和细胞周期。

关键词： colorectal cancer   ixazomib   differentially expressed genes   functional enrichment analysis   protein-protein interaction   PROTEIN-INTERACTION NETWORKS   C-JUN   DNA-DAMAGE   IN-VITRO   APOPTOSIS   CELLS   ATF3   EXPRESSION   INDUCTION   GENE  

摘要： Purpose: The study aimed to explore the anticancer effects of a novel proteasome inhibitor, ixazomib, on colorectal cancer (CRC) using a combined method of microarray and bioinformatics analysis. Materials and methods: Cell proliferation was tested by Cell Counting Kit-8 (CCK-8) assay for SW620 cells treated with different concentrations of ixazomib and different treatment times. The microarray analysis was conducted for six samples, including three samples of SW620 cells untreated with ixazomib and three samples of SW620 cells treated with ixazomib. The differentially expressed genes (DEGs) between untreated and treated samples were identified by the Linear Models for Microarray data (LIMMA) package in R language. The Gene Ontology (GO) functional and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed for the DEGs using the Database for Annotation, Visualization and Integrated Discovery (DAVID) and KEGG Orthology-Based Annotation System (KOBAS) online tool. The protein-protein interaction (PPI) network was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, and module analysis was performed for the PPI network. Results: Ixazomib could inhibit the proliferation of SW620 cells in a dose-dependent and time-dependent manner. A total of 743 DEGs, including 203 upregulated DEGs such as HSPA6 and 540 downregulated DEGs such as APCDD1, were identified. Eighty-three GO terms were enriched for DEGs, which were mainly related to protein folding, apoptotic process, transcription factor activity, and proteasome. Thirty-seven KEGG pathways were perturbed, including pathway of apoptosis and cell cycle. Forty-six hub genes, such as TP53, JUN, and ITGA2, were screened out, and three modules with important functions were mined from the PPI network. Conclusion: The novel proteasome inhibitor ixazomib significantly inhibited the proliferation of human CRC SW620 cells. It exerted anticancer effects