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| 1 | High levels of homocysteine downregulate apolipoprotein E expression via nuclear factor kappa B显示文摘AIM: To investigate the effect of high homocysteine(Hcy) levels on apolipoprotein E(apoE) expression and the signaling pathways involved in this gene regulation.METHODS: Reverse transcriptase polymerase chain reaction(RT-PCR) and Western blot were used to assess apo E expression in cells treated with various concentrations(50-500 μmol/L) of Hcy. Calcium phosphatetransient transfections were performed in HEK-293 and RAW 264.7 cells to evaluate the effect of Hcy on apoE regulatory elements [promoter and distal multienhancer 2(ME2)]. To this aim, plasmids containing the proximal apoE promoter [(-500/+73)apoE construct] alone or in the presence of ME2 [ME2/(-500/+73)apoE construct] to drive the expression of the reporter luciferase gene were used. Co-transfection experiments were carried out to investigate the downstream effectors of Hcymediated regulation of apoE promoter by using specific inhibitors or a dominant negative form of IKβ. In other co-transfections, the luciferase reporter was under the control of synthetic promoters containing multiple specific binding sites for nuclear factor kappa B(NF-κB), activator protein-1(AP-1) or nuclear factor of activated T cells(NFAT). Chromatin immunoprecipitation(ChI P)assay was accomplished to detect the binding of NF-κB p65 subunit to the apoE promoter in HEK-293 treated with 500 μmol/L Hcy. As control, cells were incubated with similar concentration of cysteine. NF-κB p65 proteins bound to DNA were immunoprecipitated with anti-p65 antibodies and DNA was identified by PCR using primers amplifying the region-100/+4 of the apoE gene. RESULTS: RT-PCR revealed that high levels of Hcy(250-750 μmol/L) induced a 2-3 fold decrease in apoE m RNA levels in HEK-293 cells, while apo E gene expression was not significantly affected by treatment with lower concentrations of Hcy(100 μmol/L). Immunoblotting data provided additional evidence for the negative role of Hcy in apoE expression. Hcy decreased apoE promoter activity, in the presence or absence of ME2, in a dose dependent manner, in both RAW 264.7 and HEK-293 cells, as revealed by transient transfection experiments. The downstream effectors of the signaling pathways of Hcy were also investigated. The inhibitory effect of Hcy on the apo E promoter activity was counteracted by MAPK/ERK kinase 1/2(MEK1/2) inhibitor U0126, suggesting that MEK1/2 is involved in the downregulation of apoE promoter activity by Hcy. Our data demonstrated that Hcy-induced inhibition of apoE took place through activation of NF-κB. Moreover, we demonstrated that Hcy activated a synthetic promoter containing three NF-κB binding sites, but did not affect promoters containing AP-1 or NFAT binding sites. ChI P experiments revealed that NF-κB p65 subunit is recruited to the apoE promoter following Hcy treatment of cells.CONCLUSION: Hcy-induced stress negatively modulates apoE expression via MEK1/2 and NF-κB activation. The decreased apo E expression in peripheral tissues may aggravate atherosclerosis, neurodegenerative diseases and renal dysfunctions. | Violeta G Trusca Adina D Mihai Elena V Fuior Ioana M Fenyo Anca V Gafencu | 2016 | World Journal of Biological Chemistry2016，7，1： | 6 |
| 2 | Confocal laser endomicroscopy and immunoendoscopy for real-time assessment of vascularization in gastrointestinal malignancies显示文摘Gastrointestinal cancers represent a major cause of morbidity and mortality,with incomplete response to chemotherapy in the advanced stages and poor prognosis.Angiogenesis plays a crucial part in tumor growth and metastasis,with most gastrointestinal cancers depending strictly on the development of a new and devoted capillary network.Confocal laser endomicroscopy is a new technology which allows in vivo microscopic analysis of the gastrointestinal mucosa and its microvascularization during ongoing endoscopy by using topically or systemically administered contrast agents.Targeting markers of angiogenesis in association with confocal laser endomicroscopic examination(immunoendoscopy),as a future challenge,will add functional analysis to the morphological aspect of the neoplastic process.This review describes previous experience in endomicroscopic examination of the upper and lower digestive tract with emphasis on vascularization,resulting in a broad spectrum of potential clinical applications,and also preclinical research that could be translated to human studies. | Dan Ionut Gheonea Tatiana Crtnǎ Tudorel Ciurea Carmen Popescu Anca Bǎdǎrǎu Adrian Sǎftoiu | 2011 | World Journal of Gastroenterology2011，17，1： | 6 |
| 3 | Dynamic modelling of wind farm grid interaction显示文摘 | ANCA D POUI S FREDE B | 2002 | Wind engineering2002，26，4： | 1 |
| 4 | Initialisation of grid-connected wind turbine model in power-system simulations显示文摘 | Anca D H Poul S Florin I | 2003 | Wind Engineering2003，27，1： | 1 |
| 5 | Centralized power control of wind farm with doubly fed induction generators显示文摘 | Anca D H Poul S Florin I | | 0，，07： | 1 |
| 6 | A peroxidase gene promoter induced by phytopathogens and Methyl Jasmonate in transgenic plant显示文摘 | Mark D C Anne L R Anca G R | 1997 | Molecular Plant-Microbe Interactions1997，10，： | 1 |
| 7 | Simulation Model of an Active-Stall Fixed-Speed Wind Turbine Controller显示文摘 | CLEMENS J ANCA D H POUL S | | 0，，02： | 1 |
| 8 | Wind turbine concept market pene- tration over 10 years ( 1995-2004 ) 显示文摘 | Anca D H Lars H H | 2007 | Wind Energy2007，10，1： | 1 |
| 9 | Fault Ride-through Ca- pability of DFIG Wind Turbines 显示文摘 | Anca D Hansena Gabriele Michalke | 2007 | Renewable Energy2007，32，9： | 1 |
| 10 | Dynamic Modelling of Wind Farm Grid Interaction显示文摘 | Anca D Poul S Frede B | 2002 | Wind engineering2002，26，4： | 1 |
| 11 | Tailoring the composition and properties of sprayed CuSbS2 thin films by using polymeric additives显示文摘 | Ionut P Anca D | 2012 | International Journal of Photoenergy2012，2012，： | 1 |
| 12 | Quality of life in stroke survivors after local intra-arterial thrombolysis显示文摘 | Fischer U Anca D Arnold M | 2008 | Cerebrovasc Dis2008，25，5： | 1 |
| 13 | A peroxidase gene promoter induced by phytopathogens and Methyl Jasmonate in transgenic plant 显示文摘 | Mark Curtis D Anne Rae L Anca Rusu G | 1997 | Molecular Plant Microbe interactions1997，，10： | 1 |
| 14 | Centralized power control of wind farm with doubly fed induction generators显示文摘 | Anca D H Poul S Florin I | 2006 | Renewable Energy2006，31，7： | 1 |
| 15 | Quality of life in stroke survivors after local intra-arterial thrombolysis显示文摘 | Fischer U Anca D Arnold M | 2008 | Cerebrovasc Dis2008，25，5： | 1 |
| 16 | AGEs and glucose levels modulate type Ⅰ and Ⅲ procollagen mRNA synthesis in dermal fibroblasts cells culture显示文摘 | Andreea SI Marieta C Anca D | 2008 | Exp Diabetes Res2008，2008，： | 1 |
| 17 | AGEs and glucoselevels modulate type I and III procollagen mRNA synthesis in dermalfibroblasts cells culture显示文摘 | ANDREEA S I MARIETA C ANCA D | 2008 | Experimental Diabetes Research2008，2008，： | 1 |
| 18 | Progress in the field of conducting polymers for tissue engineering applications显示文摘 | Anca D B Luminita C Ioan C | 2011 | J Biol Appl2011，26，7： | 1 |
| 19 | Wind turbine concept market penetration over 10 years (1995 - 2004)显示文摘 | Hansen Anca D Hansen Lars H | 2007 | Wind Energy2007，10，1： | 1 |
| 20 | Wind turbine concept market penetration over 10 years (1995- 2004)显示文摘 | Anca D Hansen Lars H Hansen | 2007 | Wind Energy2007，10，1： | 1 |
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