关键词： CORONARY heart disease treatment   VENTRICULAR remodeling   HEART cells  

关键词： HEART cells   MACROPHAGES   TRANSCRIPTION factors  

关键词： ATF3   elevated hydrostatic pressure   apoptosis   glaucoma  

摘要： AIM：To investigate the effect of activating transcription factor-3（ATF3）-deletion on apoptosis of cultured retinal ganglion cells（RGCs）.METHODS：Three ATF3 siR NA（ATF3-rat-651, ATF3-rat-319, ATF3-rat-520） were constructed, and were transiently transfected into RGC-5 cells. Quantitative real-time polymerase chain reaction（PCR） was used to examine ATF3 expression and the most effective ATF3 siR NA was selected for further studies. Flow cytometry was applied to investigate the effects of ATF3 deletion on RGC-5 apoptosis under elevated hydrostatic pressure. Quantitative realtime PCR and Western blot were performed to validate differentially expressed genes and proteins in ATF3-knockdown RGC-5 ***：ATF3 specific siR NA effectively down-regulated ATF3 expression and significantly inhibited cell apoptosis in RGC-5 cells. Quantitative real-time PCR and Western blot confirmed that ATF3 knockdown remarkably decreased Jun-B and increased c-Jun at both m RNA and protein levels in RGC-5 ***：ATF/cA MP-response element-binding family of transcription factors may be involved in the development of glaucoma and could be novel treatment targets for glaucoma.

关键词： Apoptosis   Cell death   Alpha-lactalbumin   Histone deacetylase inhibition   Involution NF-kappa B  

摘要： alpha-lactalbumin is a protein of dual function found in milk of most mammals. alpha-lactalbumin binds beta-1,4galactosyltransferase to form the regulatory subunit for lactose synthesis and has also been shown to cause cell death. This study shows, for the first time, that alpha-lactalbumin isolated in a rare 28 kDa dimeric form induces cell death, while 14 kDa monomeric alpha-lactalbumin is inactive. In contrast to the casein derived and chemically induced alpha-lactalbumin variants, MAL and HAMLET/BAMLET, the effects of 28 kDa alpha-lactalbumin are calcium independent and, unlike MAL and HAMLET, 28 kDa alpha-lactalbumin dimer causes cell death of primary mammary cells and a variety of immortalised cell lines, which are committed to cell death pathways within 1-4 h of exposure. Microarray analysis confirmed that cell death was the result of an apoptotic response. Functional assays determined that the mechanism by which 28 kDa alpha-lactalbumin kills cells involved inhibition of histone deacetylase activity mediated by NF-kB. We also show that 28 kDa alpha-lactalbumin occurs naturally in the milk of cows, goats and sheep, is low in concentration during mid-lactation, but accumulates during milk stasis, consistent with a role in involution. (C) 2017 Elsevier Inc. All rights reserved.

关键词： Insulin-like growth factor-1   Tumor necrosis factor-alpha   Activating transcription factor 3   Growth-associated protein 43   Neurite outgrowth   Neuron   Dorsal root ganglion  

摘要： Upregulation of the pro-inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) is involved in the development and progression of numerous neurological disorders. Recent reports have challenged the concept that TNF-alpha exhibits only deleterious effects of pro-inflammatory destruction, and have raised the awareness that it may play a beneficial role in neuronal growth and function in particular conditions, which prompts us to further investigate the role of this cytokine. Insulin-like growth factor-1 (IGF-1) is a cytokine possessing powerful neuroprotective effects in promoting neuronal survival, neuronal differentiation, neurite elongation, and neurite regeneration. The association of IGF-1 with TNF-alpha and the biological effects, produced by interaction of IGF-1 and TNF-alpha, on neuronal outgrowth status of primary sensory neurons are still to be clarified. In the present study, using an in vitro model of primary cultured rat dorsal root ganglion (DRG) neurons, we demonstrated that TNF-alpha challenge at different concentrations elicited diverse biological effects. Higher concentration of TNF-alpha (10 ng/mL) dampened neurite outgrowth, induced activating transcription factor 3 (ATF3) expression, reduced growth-associated protein 43 (GAP-43) expression, and promoted GAP-43 and ATF3 coexpression, which could be reversed by IGF-1 treatment;while lower concentration of TNF-alpha (1 ng/mL) promoted neurite sprouting, decreased ATF3 expression, increased GAP-43 expression, and inhibited GAP-43 and ATF3 coexpression, which could be potentiated by IGF-1 supplement. Moreover, IGF-1 administration restored the activation of Akt and p70 S6 kinase (S6K) suppressed by higher concentration of TNF-alpha (10 ng/mL) challenge. In contrast, lower concentration of TNF-alpha (1 ng/mL) had no significant effect on Akt or S6K activation, and IGF-1 administration activated these two kinases. The effects of IGF-1 were abrogated by phosphatidylinositol 3-kinase (PI3K) inhibitor

关键词： ATF3   lung cancer   tumor progression  

摘要： BackgroundLung cancer remains the most common cause of cancer-related death, with high rates of recurrence and poor outcomes. An abnormally high expression of activating transcription factor 3 (ATF3) in various cancers suggests an oncogenic role;however, its function in lung cancer is largely unknown. MethodsSixty-four pairs of lung cancer tissues were collected for ATF3 expression analysis by quantitative real-time PCR, immunoblotting, and immunohistochemistry staining. Correlations between ATF3 expression with clinicopathological features and overall survival were analyzed. ATF3 expression in a panel of lung cancer cell lines together with normal bronchial epithelial Beas-2B cells was also determined. Human H1299 and A549 cells were used for ATF3 knockdown and/or overexpression assays. Alterations in cell proliferation, cell cycle attribution, migration, and invasion were all assessed in vitro. ResultsIncreased ATF3 messenger RNA and protein expression were observed in lung cancer tissues/cells compared with normal tissues/cells. High tumorous ATF3 expression was significantly correlated with positive advanced tumor grade, lymph node metastasis, and shorter overall survival. Experimentally, we found that RNA interference mediated knockdown of ATF3 significantly inhibited the cell proliferation, cell cycle progression, migration, and invasion capacities of lung cancer cells in vitro, whereas forced expression of ATF3 did the opposite. ConclusionUpregulation of ATF3 in lung cancer promotes cell proliferation, migration, and invasion, and may represent a novel therapeutic target for lung cancer.

关键词： 2-OhβCD 2-hydroxy-beta-cyclodextrin   ATF3   ATF3 activating transcription factor 3   CEBP CCAAT/ enhancer binding protein   Cell signaling   DAMPs damage-associated molecular pattern molecules   ERK extracellular-signal-regulated kinase   HAECs human aortic endothelial cells   HDL   IL-1β interleukin 1 beta   IL-6 interleukin 6   IL-8 interleukin 8   Inflammation   JNK N-terminal kinase   LPS lipopolysaccharide   MβCD methyl-beta-cyclodextrin   MAP kinase   MAPKs mitogen-activated protein kinases   MIP-1α macrophage inflammatory protein 1 alpha   NF-κB nuclear factor kappa B   PA palmitic acid   PI3K Phosphoinositide 3-kinase   PKC protein kinase C   pPCR quantitative real-time PCR   Saturated fatty acid   TLRs Toll-like receptors  

摘要： Background and aims Peripheral insulin resistance is associated with several metabolic abnormalities, including elevated serum fatty acids that contribute to vascular injury and atherogenesis. Our goals were to examine whether saturated fatty acids can modify innate immune responses to subclinical concentrations of lipopolysaccharide (LPS) in endothelial cells, and to explore the underlying pathway and determine whether it is modified by high density lipoprotein (HDL) and other factors commonly altered in insulin resistance. Methods Physiologic concentrations of palmitic acid were added to human aortic endothelial cells with and without a variety of inhibitors or HDL and measures of cell inflammation and function assessed. Results Palmitic acid significantly amplified human aortic endothelial cell inflammatory responses to LPS. Similar results were obtained from lipolysis products of triglyceride rich lipoproteins. Metabolism of palmitic acid to ceramide and subsequent activation of PKC-ζ, MAPK and ATF3 appeared critical in amplifying LPS induced inflammation. The amplified response to palmitic acid/LPS was decreased by HDL, dose dependently, and this inhibition was dependent on activation of PI3K/AKT and reduction in ATF3. Conclusions These results indicate that endothelial cell innate immune responses are modified by metabolic abnormalities commonly present in insulin resistance and provide evidence for a novel mechanism by which HDL may reduce vascular inflammation. [ABSTRACT FROM AUTHOR]

关键词： Saturated fatty acid   HDL   Inflammation   Cell signaling   MAP kinase   ATF3  

摘要： Background and aims: Peripheral insulin resistance is associated with several metabolic abnormalities, including elevated serum fatty acids that contribute to vascular injury and atherogenesis. Our goals were to examine whether saturated fatty acids can modify innate immune responses to subclinical concentrations of lipopolysaccharide (LPS) in endothelial cells, and to explore the underlying pathway and determine whether it is modified by high density lipoprotein (HDL) and other factors commonly altered in insulin resistance. Methods: Physiologic concentrations of palmitic acid were added to human aortic endothelial cells with and without a variety of inhibitors or HDL and measures of cell inflammation and function assessed. Results: Palmitic acid significantly amplified human aortic endothelial cell inflammatory responses to LPS. Similar results were obtained from lipolysis products of triglyceride rich lipoproteins. Metabolism of palmitic acid to ceramide and subsequent activation of PKC-zeta, MAPK and ATF3 appeared critical in amplifying LPS induced inflammation. The amplified response to palmitic acid/LPS was decreased by HDL, dose dependently, and this inhibition was dependent on activation of PI3K/AKT and reduction in ATF3. Conclusions: These results indicate that endothelial cell innate immune responses are modified by metabolic abnormalities commonly present in insulin resistance and provide evidence for a novel mechanism by which HDL may reduce vascular inflammation. (C) 2016 Published by Elsevier Ireland Ltd.

关键词： Drosophila   Innate immunity   Intestinal stem cells  

摘要： Epithelial barrier function is maintained by coordination of cell proliferation and cell loss, whereas barrier dysfunction can lead to disease and organismal death. JNK signalling is a conserved stress signalling pathway activated by bacterial infection and tissue damage, often leading to apoptotic cell death and compensatory cell proliferation. Here we show that the stress inducible transcription factor ATF3 restricts JNK activity in the Drosophila midgut. ATF3 regulates JNK-dependent apoptosis and regeneration through the transcriptional regulation of the JNK antagonist, Raw. Enterocyte-specific ATF3 inactivation increases JNK activity and sensitivity to infection, a phenotype that can be rescued by Raw overexpression or JNK suppression. ATF3 depletion enhances intestinal regeneration triggered by infection, but does not compensate for the loss of enterocytes and ATF3-depleted flies succumb to infection due to intestinal barrier dysfunction. In sum, we provide a mechanism to explain how an ATF3-Raw module controls JNK signalling to maintain normal intestinal barrier function during acute infection.

关键词： RNA干扰   p300   亚溶解型C5b-9   凋亡   乙酰化  

摘要： 目的:构建大鼠p300基因短发夹状小干扰RNA(short hairpin RNA,shRNA)真核表达质粒,观察沉默p300基因对亚溶解型(sublytic)C5b-9诱导大鼠肾小球系膜细胞(glomerular messangial cell,GMC)凋亡和激活转录因子3(activating transcription factor 3,ATF3)乙酰化水平的影响。方法:用DNA重组技术针对p300基因不同位点设计3个shRNA序列,然后分别克隆到真核表达载体pGCsi-U6/Neo/GFP/shRNA中,经电转染入GMC,再给予sublytic C5b-9刺激。Western blot筛选干涉效果最佳的p300 shRNA,流式细胞术检测GMC凋亡率,免疫沉淀(immunoprecipitation,IP)和Western blot联合检查ATF3乙酰化水平。结果:核酸测序表明p300 shRNA重组质粒构建成功。Western blot显示p300 shRNA-2具有最佳沉默效率。p300 shRNA处理GMC后,由sublytic C5b-9诱导的GMC凋亡率显著下降,同时ATF3乙酰化水平也明显降低。结论:成功构建了大鼠p300 shRNA真核表达质粒,并初步证实了p300能够促进sublytic C5b-9诱导的GMC凋亡,其机制可能与p300乙酰化修饰ATF3有关。