TRPV4 Initiates Ventilator Induced Lung Injury

TRPV4 引发呼吸机引起的肺损伤

基本信息

批准号：
7851392
负责人：
JAMES Courtney PARKER
金额：
$ 35.61万
依托单位：
UNIVERSITY OF SOUTH ALABAMA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7851392
关键词：
3-nitrotyrosine Acute Lung Injury Adult Respiratory Distress Syndrome Agonist Alveolar Alveolar Macrophages Arachidonic Acids Back Blood Vessels Blood capillaries Breathing Calcium Calcium Channel Cations Cells Clinical Research Clinical Trials Coagulation Process Coculture Techniques Cyclic AMP-Dependent Protein Kinases Cyclic GMP Cysteine Cytochrome P450 Data Development Dose Endothelial Cells Endothelium Environmental air flow Epithelial Epithelial Cells F2R gene Fluorescence Microscopy Gene Deletion Genotype Heating Inflammatory Response Infusion procedures Injury Intervention Ion Channel Laboratories Lung Lung Inflammation Measures Mechanical ventilation Mechanics Mediating Mus Neurons Nitric Oxide Nitric Oxide Donors Nitrogen Oxygen PAR-2 Receptor Patients Peptide Hydrolases Peptides Permeability Peroxonitrite Phagocytosis Pharmacological Treatment Phorbol Phorbol Esters Phorbols Production Protein Tyrosine Phosphatase Proteins Pulmonary Edema Rattus Reporting Research Rest Role Stretching Temperature Testing Thrombin Tidal Volume Trypsin Tyrosine Tyrosine Phosphorylation Vanilloid Vascular Permeabilities Ventilator-induced lung injury activated Protein C anandamide capillary electric impedance feeding inhibitor/antagonist macrophage migration monolayer mortality patch clamp pressure prevent protein expression receptor response src-Family Kinases

项目摘要

After some 40 years of research and numerous failed clinical trials, only two interventions have been shown to significantly reduce the high mortality rate of the acute respiratory distress syndrome (ARDS), a reduction of tidal volume during mechanical ventilation (22%), and infusion of activated protein C (19%). These clinical studies indicate that coagulation activation renders the lung exquisitelv sensitive to stretch because the injurious lung distention producing a 22% increase in ARDS fatalities is half that in resting tidal breathing. We recentty reported that experimental Ventilator Induced Lung Injury was prevented in mice by deletion of the calcium-permeable stretch-activated transient receptor potential vanilloid-4 (TRPV4) ion channel. Recent studies indicate that the proteinase activated receptor-2 (PAR2) associates with the TRPV4 channel in neurons and greatly amplifies the TRPV4 induced calcium increase. We show (Preliminary Data) that lung epithelial cell monolayers co-cultured with alveolar macrophages activated with trypsin (PAR2), thrombin (PAR1, 3, 4) or the TRPV4 agonist, 4alpha-phoroot 12,13--didecanoate (4a-PDD) given separately actually decreased monolayer permeability, but that pre-treatment with either trypsin or thrombin followed by 4a-PDD produced a sustained permeability increase. We Hvpothesize: that coagulation induced proteases and TRPV4 induced alveolar macrophage release of proteases that activate PAR2 and PAR1, 3, and 4 which then amplify the mechanogated TRPV4 response in epithelial and endothelial cells sufficiently to cause increased lung vascular permeability. We propose these Specific Aims: 1) Test the hypotheses that: PAR1 and PAR2 activation peptides followed by 4a-PDD amplify the TRPV4 mediated calcium influx in alveolar macrophages and alveolar epithelial cells measured using whole cell patch clamp and fluorescence microscopy; and that the PAR amplification is reduced by inhibitors of PLC~, PKA, and PKC. 2) Test the hypothesis that PAR1 and PAR2 activation peptides followed by 4a-PDD increases alveolar epithelial monolayer permeability using electric cell-substrate impedance sensing, and that lung PAR1 and PAR2 activation peptidesincrease lung vascular permeability in TRPV4+/+ mice ventilated with high airway pressures but not in TRPV4-/- mice, or either genotype ventilated with low airway pressures

经过大约40年的研究和许多失败的临床试验，只有两种干预措施显示可显着降低急性呼吸窘迫综合征的高死亡率（ARDS），机械通气期间潮汐体积的减少（22％）和激活的输注蛋白C（19％）。这些临床研究表明凝血激活使肺大优先型对拉伸敏感，因为有害的肺延伸会增加22％ ARDS死亡是静止潮气的一半。我们最近报道了实验性呼吸机通过删除钙渗透的拉伸激活而预防小鼠诱导的肺损伤瞬态受体电势香草素-4（TRPV4）离子通道。最近的研究表明蛋白酶激活的受体-2（PAR2）与神经元中的TRPV4通道相关，并且很大放大TRPV4诱导的钙增加。我们显示（初步数据）肺上皮细胞单层与胰蛋白酶（PAR2），凝血酶（PAR1、3， 4）或trpv4激动剂，4Alpha-Phoroot 12,13-二足（4A-PDD）实际上分别给定单层渗透性降低，但使用胰蛋白酶或凝血酶的预处理，然后是 4A-PDD产生了持续的渗透率。我们hvpothess：凝血引起蛋白酶和TRPV4诱导肺泡巨噬细胞释放，激活PAR2 然后，PAR1，3和4，然后在上皮和内皮细胞充分引起肺血管渗透性增加。我们提出了这些具体目的：1）检验：PAR1和PAR2激活肽，然后是4A-PDD的假设扩增肺泡巨噬细胞和肺泡上皮细胞中TRPV4介导的钙涌入使用全细胞斑块夹和荧光显微镜测量；并放大 PLC〜，PKA和PKC的抑制剂降低。 2）检验PAR1和PAR2的假设激活肽随后4A-PDD增加了使用使用的肺泡上皮单层渗透率电细胞基底阻抗感应，肺PAR1和PAR2激活肽酶 TRPV4+/+小鼠的肺血管渗透性以高气道压力通风，但不在 TRPV4 - / - 小鼠，或用低气道压力通风的基因型