关键词： 转录激活因子3   苔藓纤维出芽   癫痫  

摘要： 目的通过观察戊四氮(pentylenetetrazole; PTZ)大鼠点燃模型海马CA3区及齿状回区转录激活因子3(activating transcription factor3; ATF3)的动态表达变化,结合海马苔藓纤维出芽(mossy fiber sprouting; MFS)的程度,探讨其与MFS的关系及在癫痫过程中的作用。 方法健康SD雄性成年大鼠150只,随机分为生理盐水对照组(60只)和PTZ实验组(90只)。实验组每日腹腔注射PTZ (30mg/kg)；同时生理盐水组注射等量生理盐水。按照首次腹腔注射后第3天,1周,2周,4周和6周的时间点分配,将PTZ组及生理盐水组随机分为5个亚组。根据实验方法的不同再将每个亚组分为三个小组：第一组用于免疫组化及免疫荧光,检测各时间点ATF3蛋白在海马各区的动态表达及在细胞内表达定位；第二组用于Western Blot,将海马各区ATF3表达变化进行半定量分析；第三组用于Timm染色,检测苔藓纤维出芽情况,其中实验组每小组各6只,对照组每小组各4只。 结果 1.动物行为学表现 大鼠多在连续注射PTZ21-28天(平均23.7±1.9天)后被点燃。模型点燃率为94.4%,死亡率为5.33%。除了4周和6周组各有一只大鼠未出现痫性发作外,点燃成功后大鼠出现诱发发作及自发发作,对照组未出现痫性发作。 2.苔藓纤维出芽评分 实验组CA3区与对照组相对应时间点相比MFS评分均有显著性差异(P<0.05)。从3天起评分逐渐增加,4周时达高峰,至6周仍维持在较高水平。齿状回区MFS评分在实验组与对照组之间以及各组内不同时间点之间比较均无显著性差异(P>0.05)。 ***3蛋白的动态表达 免疫组化示在癫痫点燃过程中,实验组：海马CA3区ATF3蛋白表达逐渐上升,在4周组达最高峰,6周组稍有下降(P<0.05)；齿状回区ATF3蛋白在点燃过程中无明显变化(P>0.05)。对照组：海马CA3区及齿状回区在3天到6周组均无明显变化(P>0.05)。Western Blot结果示海马ATF3的表达自3天组逐渐上升,至4周组达高峰,6周组较前明显下降；除了3天组之外,其他各时间点ATF3表达在实验组均较对照组高(P<0.05)。免疫荧光结果示ATF3蛋白定位于大鼠海马神经元细胞核及细胞浆中,且主要集中在细胞浆。 结论 ATF3蛋白在海马CA3区的表达上调可能促进苔藓纤维出芽,进而导致颞叶癫痫的发生及发展。

关键词： ADRBK1   AKT3   ATF3   DIO2   rSNPs   TBXA2R and VEGFA genes   transcriptional factor binding site   transcriptional factors  

摘要： Regulatory single nucleotide polymorphisms (rSNPs) which change the transcriptional factor binding sites (TFBS) for transcriptional factors (TFs) to bind DNA were reviewed for the ADRBK1 (GRK2), AKT3, ATF3, DIO2, TBXA2R and VEGFA genes. Changes in the TFBS where TFs attach to regulate these genes may result in human sickness and disease. The highlights of this previous work were reviewed for these genes. [ABSTRACT FROM PUBLISHER]

关键词： 兔椎间盘退变模型   等离子汽化消融术   TNF-α   ATF3   GAP43  

摘要： 目的：研究等离子汽化消融术兔退变椎间盘内TNF-α和背根神经节内ATF3、GAP43神经元表达的影响，探索其治疗盘源性腰痛的机制；在不同椎间盘模型内的差异表达，探索盘源性腰痛发病机制。 方法：健康新西兰大白兔，不计雌雄72只，体质量2.5-3.5kg，排除受孕及患病兔，采用针刺法制造新西兰大白兔L5-6椎间盘退变模型，所有实验动物学均在不同阶段进行椎间盘内注入神经示踪剂-荧光金（Fluoro-Gold），根据椎间盘退变和等离子汽化消融术两因素进行析因分析设计分组，依据椎间盘退变因素随机分为两组A组（椎间盘退变组）和B组（正常椎间盘组），然后以等离子汽化消融术因素随机分为治疗组（A1组或B1组）对照组（A2组或B2组，假手术组），收集不同时间段的椎间盘标本和背根神经节标本，免疫组化检查椎间盘髓核内TNF-α和背根神经节内ATF3、GAP43阳性神经元数目。统计分析采用SPSS17.0软件进行,检验水准α＝0.05，P＜O.05认为差异有统计学意义。 结果：正常组治疗组术后1天椎间盘组织中TNF-α的活性为90.23±5.56，术后1周为88.45±4.37，术后2周87.33±5.84；正常组治疗组术后1天椎间盘组织中ATF3、GAP43阳性细胞计数分别为2.73±1.74、2.62±1.63，术后1周为2.54±1.51、2.51±1.49，术后2周2.28±1.27、2.25±1.20；正常组对照组术后1天TNF-α的活性为91.18±4.36，术后1周为91.78±5.25，术后2周93.33±5.45；正常组对照组术后1天ATF3、GAP43阳性细胞计数为3.05±1.44、3.02±1.35，术后1周为3.21±1.49、3.16±1.35，术后2周3.35±1.78、3.29±1.62；模型治疗组消融术后1天椎间盘组织内TNF-α活性为184.78±5.43，术后1周为154.32±4.66，术后2周为141.46±8.52；模型治疗组消融术后1天椎间盘组织内ATF3、GAP43阳性细胞计数为5.63±1.46、5.55±1.37，术后1周为5.12±1.42、5.04±1.34，术后2周为4.65±1.34、4.56±1.26；模型对照组相应时间点椎间盘组织内TNF-α活性分别为195.78土10.82，196.43±12.80，225.76±11.38；模型对照组相应时间点椎间盘组织内ATF3、GAP43阳性细胞计数分别为6.23土1.32，6.19土1.26；6.45土1.36，6.39土1.28；7.32±1.68，7.28±1.61。TNF-α活性、ATF3、GAP43阳性细胞计数:模型组各时间点均较正常组明显升高(P0.05)。 结论：等离子汽化消融术可以降低兔退变椎间盘内TNF-α活性及相关神经的增生，是治疗椎间盘源性腰痛的机制之一。盘源性腰痛与TNF-α等炎症因子及ATF3，GAP43的表达有关。

关键词： ATF3   rSNPs   TFBS   Human   Etiology  

摘要： Three rSNPs (rs3125289, rs1877474 and rs11119982) in one intron of the activating transcription factor 3 (ATF3) gene have been significantly associated with the human etiology of hypospadias and may be associated with human disease. These rSNP alleles alter the DNA landscape for potential transcriptional factors (TFs) to attach resulting in changes in transcriptional factor binding sites (TFBS). These TFBS changes are examined with respect to the human etiology of hypospadias which has been found to be significantly associated with the rSNPs.

关键词： Cytokine Gene   ATF3 gene   NOD2 gene   NF-kappa B   Cytokines  

摘要： Regulation of microbially induced cytokine secretion is critical in intestinal immune homeostasis. NOD2, the Crohn's diseaseassociated bacterial peptidoglycan sensor, activates the NF-kappa B pathway. After chronic NOD2 stimulation in human macrophages, cytokine secretion is significantly attenuated, similar to the situation in the intestinal environment. We find that NF-kappa B1 (p105/p50) expression is upregulated with chronic NOD2 stimulation and is required for attenuation of cytokine secretion in vitro in human macrophages and in vivo in mice. Upon chronic NOD2 stimulation, regulation of both activating (H3K4Me2 and H4Ac) and inhibitory (H3K27Me3) histone modifications was observed within cytokine gene promoters;these outcomes were NF-kappa B1 dependent. In addition to enhanced binding to cytokine gene promoters with chronic NOD2 stimulation, NF-kappa B1 bound to the promoter of the transcriptional repressor, ATF3. ATF3 was then induced and bound to cytokine gene promoters;both features were impaired upon NF-kappa B1 knockdown. Restoring ATF3 expression under NF-kappa B1 knockdown conditions restored NOD2-mediated cytokine downregulation. Finally, NF-kappa B1 and ATF3 cooperate with other inhibitory pathways, including IRAKM and secreted mediators, to downregulate cytokine secretion after chronic NOD2 stimulation. Therefore, we identify NF-kappa B1 and ATF3 as critical mechanisms through which NOD2 downregulates cytokines and contributes to intestinal immune homeostasis.

关键词： Chemotherapy-induced neuropathy   satellite cells   microglia   astrocytes   ATF3   neurofilament  

摘要： Neurotoxicity is the limiting side effect of the anticancer agent oxaliplatin. A tangled panel of symptoms, sensory loss, paresthesia, dysesthesia, and pain may be disabling for patients and adversely affect their quality of life. To elucidate the morphologic and molecular alterations that occur in the nervous system during neuropathy, rats were daily injected with 2.4 mg kg(-1) oxaliplatin intraperitoneally. A progressive decrease in the pain threshold and hypersensitivity to noxious and nonnoxious stimuli were evidenced during the treatment (7, 14, 21 days). On day 21, morphometric alterations were detectable exclusively in the dorsal root ganglia, whereas the activating transcription factor 3 and neurofilament (heavy-chain) expression changed dramatically in both the nerves and ganglia. Inflammatory features were not highlighted. Interestingly, satellite cells exhibited signs of activation. Glial modulation was characterized in the spinal cord and brain areas involved in pain signaling. On the 21st day, spinal astrocytes increased numerically whereas the microglial population was unaltered. The number of glial cells in the brain differed according to the zone and treatment time points. In particular, on day 21, a significant astrocyte increase was measured in the anterior cingulate cortex, somatosensory area 1, neostriatum, ventrolateral periaqueductal gray, and nucleus raphe magnus. (C) 2013 by the American Pain Society

关键词： Type 2 diabetes   Pancreatic beta-cells   ER stress   AMPK   ATF3   In vivo knockdown  

摘要： In obese Zucker diabetic fatty (ZDF) rats, ER stress is associated with insulin resistance and pancreatic beta-cell dysfunction: however the exact mechanisms by which ER stress drives type-2 diabetes remain uncertain. Here, we investigated the role of ATF3 on the preventive regulation of AMPK against ER stress-mediated beta-cell dysfunction during the end-stage progression of hyperglycemia in ZDF rats. The impaired glucose metabolism and beta-cell dysfunction were significantly increased in late-diabetic phase 19-week-old ZDF rats. Although AMPK phosphorylation reduced in 6- and 12-week-old ZDF rats was remarkably increased at 19 weeks, the increases of lipogenice genes, ATF3, and ER stress or ROS-mediated beta-cell dysfunction were still remained, which were attenuated by in vivo-injection of chemical chaperon tauroursodeoxycholate (TUDCA), chronic AICAR, or antioxidants. ATF3 did not directly affect AMPK phosphorylation, but counteracts the preventive effects of AMPK for high glucose-induced beta-cell dysfunction. Moreover, knockdown of ATF3 by delivery of in vivo-jetPEI ATF3 siRNA attenuated ER stress-mediated beta-cell dysfunction and enhanced the beneficial effect of AICAR. Our data suggest that ATF3 may play as a counteracting regulator of AMPK and thus promote beta-cell dysfunction and the development of type-2 diabetes and could be a potential therapeutic target in treating type-2 diabetes. (C) 2013 Elsevier Inc. All rights reserved.

关键词： EPITHELIAL cells   NUCLEOTIDE sequence   OLIGOMERIZATION   TRANSCRIPTION factors   T cell receptors  

摘要： In response to excessive nucleotide-binding oligomerization domain-containing protein 2 (Nod2) stimulation caused by mucosal bacterial components, gut epithelia need to activate regulatory machinery to maintain epithelial homeostasis. Activating transcription factor 3 (ATF3) is a representative regulator in the negative feedback loop that modulates TLR-associated inflammatory responses. In the current study, the regulatory effects of ribosomal stress-induced ATF3 on Nod2-stimulated proinflammatory signals were assessed. Ribosomal inactivation caused persistent ATF3 expression that in turn suppressed proinflammatory chemokine production facilitated by Nod2. Decreased chemokine production was due to attenuation of Nod2-activated NF-kappa B and early growth response protein 1 (EGR-1) signals by ATF3. However, the underlying molecular mechanisms involve two convergent regulatory pathways. Although ATF3 induced by ribosomal inactivation regulated Nod2-induced EGR-1 expression epigenetically through the recruitment of histone deacetylase 1, NF-kappa B regulation was associated with posttranscriptional regulation by ATF3 rather than epigenetic modification. ATF3 induced by ribosomal inactivation led to the destabilization of p65 mRNA caused by nuclear entrapment of transcript-stabilizing human Ag R protein via direct interaction with ATF3. These findings demonstrate that ribosomal stress-induced ATF3 is a critical regulator in the convergent pathways between EGR-1 and NF-kappa B, which contributes to the suppression of Nod2-activated proinflammatory gene expression.

关键词： GENETIC regulation   INFLAMMATION   CYTOKINES   NEISSERIA gonorrhoeae   INTERLEUKIN-6   EPITHELIAL cells   CYTOPROTECTION   APOPTOSIS  

摘要： Neisseria gonorrhoeae regulates the expression of epithelial cell genes, activates cytoprotective pathways in the infected cell and protects it from apoptosis. Many of these responses are enhanced by the Type IV pilus (Tfp). We tested the hypothesis that *** modulates the innate immune response by inducing expression of ATF3, a transcription factor that negatively regulates the expression of many cytokine genes. We further determined whether Tfp are involved in these events. We found that *** induces ATF3 expression in mucosal epithelial cells through activation of mitogen-activated protein kinases. Maximal ATF3 expression requires Tfp retraction. Knocking down endogenous levels of ATF3 results in higher levels of IL-6 transcript. Our findings strongly suggest that ATF3 is involved in suppressing cytokine expression during gonococcal infection. We propose a model for the role of ATF3 in the context of *** infection.

关键词： Integrated stress response   ATF3   Cuprizone   Amino acid  

摘要： Cuprizone [bis(cyclohexylidenehydrazide)]-induced toxic demyelination is an experimental approach frequently used to study de- and re-myelination in the central nervous system. In this model, mice are fed with the copper chelator cuprizone which leads to oligodendrocyte apoptosis and subsequent microgliosis, astrocytosis, and demyelination. The underlying mechanisms of cuprizone-induced oligodendrocyte death are still unknown. We analysed differences in amino acid levels after short-term cuprizone exposure (i.e., 4 days). Furthermore, an amino acid response (AAR) pathway activated in oligodendrocytes after cuprizone intoxication was evaluated. Short-term cuprizone exposure resulted in a selective decrease of alanine, glycine, and proline plasma levels, which was paralleled by an increase of apoptotic cells in the liver and a decrease of alanine aminotransferase in the serum. These parameters were paralleled by oligodendrocyte apoptosis and the induction of an AAR with increased expression of the transcription factors ATF-3 and ATF-4 (activating transcription factor-3 and -4). Immunohistochemistry revealed that ATF-3 is exclusively expressed by oligodendrocytes and localized to the nuclear compartment. Our results suggest that cuprizone-induced liver dysfunction results in amino acid starvation and in consequence to the activation of an AAR. We propose that this stress response modulates oligodendrocyte viability in the cuprizone animal model.