A mechanism for suppression of TNF induced endothelial dysfunction

抑制 TNF 诱导的内皮功能障碍的机制

• 批准号: 8467738
• 负责人: SHAKER A MOUSA
• 金额: $ 33.07万
• 依托单位: ALBANY COLLEGE OF PHARMACY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8467738
• 关键词: Actins; Acute; Acute Lung Injury; Adherence; Adult Respiratory Distress Syndrome; Afghanistan; Apoptosis; Biochemical; Biological Assay; Blood Vessels; Cadherins; Cell Nucleus; Cells; Cellular biology; Chronic; Complex; Cytosol; Data; Disease; Dislocations; Endothelial Cells; Endothelium; Event; Family; Functional disorder; Genetic Transcription; Genomics; Glycogen (Starch) Synthase; In Situ; In Vitro; Inflammation; Injury; Iraq; Lung; Manuscripts; Maps; Mediating; Mediator of activation protein; Membrane; Mitogen-Activated Protein Kinases; Molecular; Myosin Light Chain Kinase; Nitrogen; Nuclear; Outcome; Oxygen; Pathway interactions; Permeability; Phosphotransferases; Physiology; Predisposing Factor; Prevention; Protein Biosynthesis; Protein Isoforms; Protein Kinase C; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; Rattus; Sepsis; Septic Shock; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Syndrome; TCF Transcription Factor; Testing; Time; Translating; Translations; Trauma; Tumor Necrosis Factor Suppression; Tumor Necrosis Factor-alpha; Vascular Permeabilities; cadherin 5; combat; in vivo; in vivo Model; lung injury; monolayer; mortality; nitration; novel; operation; prevent; promoter; protein expression; public health relevance; pulmonary vascular permeability; receptor; response; response to injury

DESCRIPTION (provided by applicant): In lung microvessel endothelial cells, TNF (i.e., early ~0.5 hr) induces ONOO--mediated nitration of ¿-actin causing dislocation of ¿-catenin from adherence junctions. In our new model of in vivo vascular injury at TNF-24.0 hr, there is a "window" of suppressed vascular permeability within TNF-4.0 hr. Our new preliminary data shows, both in vitro and/or in vivo, that the suppression of (~4.0 hr) TNF-induced increased endothelial permeability is associated with nuclear ¿-catenin translocation and increases in total and membrane VE-cadherin. Our novel preliminary data demonstrates: (1) TNF-24 hr induced barrier dysfunction ex vivo is suppressed by inhibition of glycogen synthetase kinase (GSK)3a/¿ in vivo, (2) T cell factor (TCF)/ lymphoid enhancer factor (LEF)/¿-catenin dependent promoter activity is increased by TNF in vitro and in vivo, and (3) These responses to TNF are prevented by inhibition of PKC in vivo and PKCa in vitro. This proposal will test the paradigm that TNF-induced (~24.0 hr post- TNF) increase in vascular permeability is suppressed (~4.0 hr post-TNF), at least in part, by PKCa-induced inhibition of GSK3¿ activity. The decrease in GSK3¿ activity causes increased nuclear ¿-catenin. The genomic ¿-catenin-activity increases the expression of VE-cadherin. The increase in VE-cadherin expression promotes zonular-adherence junctions suppressing (i.e., braking) the initial (~0.5 hr post-TNF) increased endothelial protein permeability. The increase in VE-cadherin promotes complex formation with ¿-catenin, the off signal for continuous genomic effects of ¿-catenin. It is proposed that persistent inhibition of GSK3¿ activity will continue to suppress barrier dysfunction. We will map a novel paradigm (i.e., in vitro, in situ and in vivo) for the suppression ("braking") of TNF-induced lung injury which is GSK3¿-mediated, ¿-catenin-dependent increased VE-cadherin activity. Hypothesis: The hypothesis to be tested is TNF-induced lung injury is suppressed, at least in part by, ?PKCa ? ?GSK3¿ ? ?2-catenin ? ?VE-cadherin in adherence junctions. The Specific Aims are to determine, in vitro and/or in vivo, in endothelium that: (1) TNF causes PKCa-mediated GSK3¿-inhibition that induces nuclear ¿-catenin translocation (Years 1- 2), (2) The TNF-induced nuclear ¿-catenin translocation causes increased VE-cadherin: (I) promoter activity, (ii) RNA expression, and (iii) protein synthesis (Year 2-3), and (3) The increase in VE-cadherin protein expression suppresses, at least in part, the latter TNF-induced increase in vascular barrier dysfunction (3-4). This proposal will use rat lung microvessel endothelium and lungs isolated from rats treated in vivo. An array of biochemical, genomic, proteonomic and cell biology assays are integrated with lung physiology outcomes. This approach will translate cell-molecular pathways to the mechanisms of lung injury in vivo.

描述（由应用程序提供）：在肺微血管内皮细胞中，TNF（即早期〜0.5小时）诱导粘附的 - 肌动蛋白的ONOO介导的硝化作用，从而导致 - 粘附的脱位 - 粘附的粘附肽脱位。在我们在TNF-24.0小时处的体内血管损伤的新模型中，TNF-4.0小时内有一个抑制血管通透性的“窗口”。我们的新初步数据表明，在体外和/或体内都表明，抑制（〜4.0 hr）TNF诱导的内皮渗透性增加与核 - 核 - 钙蛋白易位以及总和膜VE-cadherin的增加有关。我们新的初步数据证明：（1）TNF -24 HR诱导的屏障功能障碍通过抑制糖原合成酶激酶（GSK）3A/¿在体内抑制3A/¿体内和（3）通过体内PKC和PKCA在体外抑制TNF的这些反应。该提案将测试TNF诱导的（TNF后TNF后24.0 HR）的范式抑制血管通透性的增加（〜4.0 HR后-TNF），至少部分是通过PKCA诱导的GSK3活动的抑制作用。 GSK3活动的减少会导致核� -Catenin增加。基因组�-Catenin活性增加了VE-钙粘蛋白的表达。 VE-钙粘蛋白表达的增加促进了抑制（即制动）最初（〜0.5 hr tnf）增加内皮蛋白渗透性的延伸性粘附连接。 VE-钙粘着蛋白的增加促进了与� -Catenin的复合形成，这是 - 帕宁蛋白连续基因组效应的OFF信号。有人提出，对GSK3活动的持续抑制作用将继续抑制障碍功能障碍。我们将绘制一个新型的范式（即体外，原位和体内），以抑制TNF诱导的肺损伤，即GSK3¿-介导的 - ®-依赖性的 - 依赖性VE-钙粘着蛋白的活性。假设：要检验的假设是否至少部分地抑制了TNF诱导的肺损伤？ ？gsk3¿？ 2-catenin？粘附连接中的ve-cadherin。 The Specific Aims are to determine, in vitro and/or in vivo, in endothelium that: (1) TNF causes PKCa-mediated GSK3¿ -inhibition that induces nuclear ¿ -catenin translocation (Years 1- 2), (2) The TNF-induced nuclear ¿ -catenin translocation causes increased VE-cadherin: (I) promoter activity, (ii) RNA expression, and (iii)蛋白质合成（2-3年）和（3）VE-钙粘蛋白蛋白表达的增加至少部分抑制了后者TNF诱导的血管屏障功能障碍的增加（3-4）。该建议将使用大鼠微血管内皮和从体内治疗的大鼠分离的肺。一系列生化，基因组，蛋白质和细胞生物学测定法与肺部生理结局融为一体。这种方法将将细胞分子途径转化为体内肺损伤机理。

项目成果

Inhibition of GSK3α/β promotes increased pulmonary endothelial permeability to albumin by reactive oxygen/nitrogen species.

GSK3α/β 的抑制通过活性氧/氮促进肺内皮细胞对白蛋白的通透性增加。

• DOI: 10.1016/j.pupt.2013.06.001
• 发表时间: 2013
• 期刊: Pulmonary pharmacology & therapeutics
• 影响因子: 3.2
• 作者: Neumann,Paul;Alsaffar,Hiba;Gertzberg,Nancy;Johnson,Arnold
• 通讯作者: Johnson,Arnold

The intracerebroventricular injection of rimonabant inhibits systemic lipopolysaccharide-induced lung inflammation.

脑室内注射利莫那班可抑制全身脂多糖诱导的肺部炎症。

• DOI: 10.1016/j.jneuroim.2015.07.001
• 发表时间: 2015
• 期刊: Journal of neuroimmunology
• 影响因子: 3.3
• 作者: Johnson,Arnold;Neumann,PaulH;Peng,Jianya;James,Janey;Russo,Vincenzo;MacDonald,Hunter;Gertzberg,Nancy;Feleder,Carlos
• 通讯作者: Feleder,Carlos