关键词： ABCA1   TLR2   ATF3   Pam3CSK4  

摘要： 目的检测ABCA1敲低对巨噬细胞中Pam3CSK4引起的炎症应答的影响。方法构建小鼠单核巨噬细胞RAW264.7的ABCA1敲低的稳定细胞系,以TLR2配体Pam3CSK4刺激该细胞系建立炎症反应细胞模型,在该细胞模型上检测相关促炎和抗炎细胞因子转录水平的表达变化。结果 ABCA1敲低的RAW264.7稳定细胞株在Pam3CSK4刺激后,IL-1β,TNF-α和IL-6表达发生显著上调(P<0.01),而转录抑制因子cAMP依赖性转录因子3(ATF3)也发生显著上调(P<0.01);与此同时,ATF蛋白家族中其它因子ATF1,ATF2,ATF4和ATF5转录水平没有发生显著变化。结论在巨噬细胞RAW264.7中,ABCA1敲低显著上调Pam3CSK4引起的促炎因子IL-1β,TNF-α和IL-6的表达的同时,也显著上调了具有抗炎效应的ATF3的表达,其对炎症反应的影响可能并非是单向的促炎作用,而是发生双向的调节,而ABCA1参与上调ATF3表达的机制可能也与其ATF蛋白家族其他成员的上游调节机制不同。

关键词： 喉癌   喉鳞状细胞癌   SOX2   ATF3  

摘要： 目的:探究喉癌组织中转录因子SOX2和ATF3的表达及其相关性。方法:运用免疫组织化学(S-P)法测定喉癌患者68例和相应癌旁组织20例中SOX2、ATF3蛋白的表达及其病理特性的联系。结果:喉癌中SOX2、ATF3阳性表达率分别为77.9%、72.1%,高于癌旁组织30%、15%(P<0.05),二者的表达与喉癌的分化程度、淋巴结转移、TNM分期有关(P<0.05),且二者在喉癌中的表达呈正相关性(r=0.380,P<0.01)。结论:SOX2和ATF3均在喉癌组织中高表达,且具有正相关性,推测二者是喉癌发生机制过程重要的生物学指标。

关键词： 尿道下裂   ATF3   包皮组织  

摘要： 目的检测散发性单纯性尿道下裂患者包皮组织中激活转录因子3(Activating transcription factor 3,ATF3)的表达情况,探索ATF3与散发性单纯性尿道下裂之间的关系。方法方便选择2014年1月—2017年1月该院收治的散发性单纯性尿道下裂患者在尿道成形术中切除的包皮组织(共35例,其中轻度10例,中度15例,重度10例)作为尿道下裂组样品,单纯包皮过长(含包茎)患者在包皮环切术中去除的多余的包皮组织(共20例)作为对照组样品。通过RT-PCR方法检测实验组和对照组ATF3 m RNA的表达情况。采用SPSS 12.0统计学软件进行数据录入,构建数据库。结果通过RT-PCR方法测得散发性单纯性尿道下裂患者包皮组织中ATF3 m RNA的相对表达量为:(0.54±0.06),对照组的包皮组织中ATF3 m RNA的相对表达量为:(0.44±0.07),从结果中可以得出在散发性单纯性尿道下裂患者的包皮组织中ATF3的表达情况比正常的尿道组织中明显增加,两组结果比较,差异有统计学意义(t=5.89,P=0.00)。比较不同严重程度中尿道下裂患者ATF3 m RNA相对表达量,不同严重程度相对表达量差异无统计学意义(F=0.13,P=0.88)。结论散发性单纯性尿道下裂患者中ATF3的表达水平明显高于正常对照组,这提示ATF3与散发性单纯性尿道下裂存在着密切的关系。为将来研究ATF3在尿道下裂中产生的相关信号转导通路提供相关实验基础,及研究尿道下裂的发病机制提供理论依据。

关键词： Sodium   Potassium   Calcium   Egrl   Atf3   Nr4a1   Ptgs2   Excitation-transcription coupling  

摘要： In rat vascular smooth muscle cells (RVSMC), 3-h Na+,K+-ATPase inhibition by ouabain or in K+-free medium resulted in the inversion of the [Na+](i)/[K+](i) ratio and elevation up to 7-fold the content of Egr1, Atf3, Nr4a1 and Ptgs2 mRNAs. Ouabain increased the rate of 45Ca(2+) influx by 2 -fold that was abolished by L-type voltage-gated Ca2+ channel blocker nicardipine, but it was resistant to Na+/Ca2+ exchanger inhibitor KB-R7943. To study the role of Ca2+-mediated signaling in the expression of Na-i(+)/K-i(+)-sensitive genes we used intracellular Ca2+ chelator BAPTA and incubated RVSMC in Ca2+-free medium. The elevation of Nr4a1 and Ptgs2 expression triggered by ouabain was diminished in Ca2+-depeleted cells as well as in the presence of nicardipine and calmodulin antagonists A-7 and W-7. Ptgs2 expression was also suppressed by inhibitor of Ca2+/calmodulin-dependent protein kinase (CaMKII) KN-93 whereas increment of Nr4a1 content triggered by ouabain was attenuated by inhibitor of Ca2+/calmodulin-dependent protein phosphatase (calcineurin, CaN) cyclosporin A. Neither Ca2+ depletion nor above listed compounds had any impact on the augmented expression of Egr1 and Atf3 in ouabain-treated RVSMC. Our results strongly suggest that dissipation of transmembrane gradient of monovalent cations increases Ptgs2 and Nr4a1 transcription via augment Ca2+ influx through L-type Ca2+ channels that, in turn, leads to CaMKII-mediated phosphorylation of CREB and calcineurin-mediated dephosphorylation of NFAT, respectively. Additional experiments should be performed to identify intermediates of Na-i(+),K-i(+)-mediated Ca2+-independent excitation-transcription coupling involved the regulation of Egr1 and Atf3 expression.

关键词： acute lung injury   ATF3   inflammation   RRID   TL1A  

摘要： Excessive inflammatory responses are critical in the pathogenesis of acute lung injury (ALI). Activating transcription factor 3 (ATF3) is a stress-induced transcriptional regulator that is a negative regulator of inflammatory responses. Therefore, we investigated the role and signaling pathways of ATF3 in lipopolysaccharide (LPS)-induced ALI in mice. The mouse macrophage RAW264.7 cells were cultured on HTS 24-Transwell filter plates in presence of ATF3 siRNA before exposure to LPS. ATF3 knock-out (KO) and wild type (WT) mice were challenged by intra-peritoneal injection of LPS (15mg/kg). Gene analysis was used to analyze differential gene expression between ATF3 KO and WT mice. LPS increased the expression of ATF3 in RAW264.7 cells and in lung tissues of mice, The concentration of TNF and IL-6 was significantly increased in ATF3 siRNA-treated RAW264.7 cells compared to control cells after LPS stimulation. The concentration of TNF, IL-6 and IL-1 in serum and lung tissue of ATF3 KO mice was significantly increased compared to ATF3 WT mice. In addition, the lung wet/dry weight and BALF protein were significantly increased in ATF3 KO mice after LPS injection at 6, 24, and 48hr. The survival of ATF3 KO mice significantly decreased. Differential gene analysis showed that TL1A was highly expressed in LPS-induced lung tissues of ATF3 KO ***, ATF3 down-regulated the expression of TL1A in RAW264.7 cells and in lung tissues. These findings suggest that ATF3 protects against LPS-induced ALI via inhibiting TL1A expression.

关键词： 转录激活因子3   消化系统肿瘤  

摘要： 转录激活因子3(activating transcription factor,ATF3)属于转录因子ATF/CREB家族的成员,是一种适应反应基因,参与多种细胞活动的调控以适应细胞内外环境的变化。近年来的研究表明,ATF3作为抑癌或致癌基因在消化道系统肿瘤的发生及演变过程中发挥重要的作用。现就ATF3在消化系统肿瘤方面的研究进展作一概述。

关键词： DNA damage response   gene expression   mRNA decay   p38   senescence   ATF3   nonsense-mediated RNA decay   persistent DNA damage  

摘要： Persistent DNA damage induces profound alterations in gene expression that, in turn, influence tissue homeostasis, tumorigenesis, and cancer treatment outcome. However, the underlying mechanism for gene expression reprogramming induced by persistent DNA damage remains poorly understood. Here, using a highly effective bioluminescence-based reporter system and other tools, we report that persistent DNA damage inhibits nonsense-mediated RNA decay (NMD), an RNA surveillance and gene-regulatory pathway, in noncycling cells. NMD suppression by persistent DNA damage required the activity of the p38 alpha MAPK. Activating transcription factor 3 (ATF3), anNMD target and a key stress-inducible transcription factor, was stabilized in a p38 alpha-and NMD-dependent manner following persistent DNA damage. Our results reveal a novel p38 alpha-dependent pathway that regulates NMD activity in response to persistent DNA damage, which, in turn, controls ATF3 expression in affected cells.

关键词： Peripheral nerve injury   Minocycline   CNTF   Colchicine  

摘要： Microglia are essential in developmental processes and maintenance of neuronal homeostasis. Experimental axotomy of motor neurons results in neurodegeneration, and microglia in motor nuclei become activated and migrate towards injured neurons. However, whether these activated microglia are protective or destructive to neurons remains controversial. In the present study, we transected the hypo-glossal nerve in BALB/c mice, causing activating transcription factor 3 (ATF3) and growth associated protein 43 (GAP43) induction, and partial neuronal death. Inhibition of microglial accumulation by minocycline administration impaired microglial accumulation, decreased GAP43 mRNA expression, and reduced motor neuron survival. Expression of ATF3 contributed to nerve regeneration, and increased within 6 h after axotomy, prior to microglial migration. Further, microglial contact with neuronal cell bodies was associated with neuronal ATF3 expression. Colchicine administration blocked lesion-induced ATF3 transcription in axotomized neurons and microglial accumulation. In addition, perineuronal microglia-derived ciliary neurotrophic factor (CNTF) increased, indicating that perineuronal microglia in the hypoglossal nucleus protect axotomized motor neurons by releasing trophic factors. We also observed that microglia secrete CNTF and that neurons have CNTFR alpha and can respond to it in vitro. CNTF promote neurite elongation and neuronal survival of primary cultured neurons. Microglia make contact through unknown neuronal signals that are possibly regulated by ATF3 in hypoglossal nucleus. Moreover, they play important roles in regenerating motor neurons and are potential new therapeutic targets for motor neuron diseases. (C) 2017 Elsevier Ltd. All rights reserved.

关键词： activating transcription factor 3   macrophage   M1/M2 polarization   tenascin-C  

摘要： There are different polarization states of macrophages, including the classically activated M1 phenotype and the alternatively activated M2 phenotype. These have different functions in the inflammation process. Activating transcription factor 3 (ATF3) is a key transcriptional regulator that inhibits the inflammatory response. However, the effects of ATF3 on migration and anti-inflammatory control mechanisms of macrophages have not been thoroughly investigated. The present study investigated the effect of ATF3 on macrophage migration and M1/M2 polarization. Results revealed that overexpression of ATF3 promoted macrophage migration and the expression of the M2 phenotype markers [cluster of differentiation (CD) 163, mannose receptor C type 1, arginase 1 and peroxisome proliferator-activated receptor gamma] and inhibited expression of the M1 phenotype markers (monocyte chemoattractant protein-1, inducible nitric oxide synthase, CD16 and tumor necrosis factor-alpha), whereas knockdown of ATF3 resulted in a contrary effect. In addition, the wingless-type MMTV integration site family member (Wnt)/beta-catenin signaling pathway was activated and the expression level of tenascin (TNC) was significantly upregulated by overexpression of ATF3. Additionally, inhibition of Wnt/beta-catenin signaling significantly attenuated the upregulatory effect of ATF3 on TNC. Finally, the effect of ATF3 on macrophage migration and markers of the M1 or M2 state was investigated using TNC-specific siRNA. In conclusion, the results of the present study suggested that ATF3 promotes macrophage migration and reverses M1-polarized macrophages to the M2 phenotype by upregulation of TNC via the Wnt/beta-catenin signaling pathway.