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Oxidant Stress Mechanisms in Preeclampsia

先兆子痫的氧化应激机制

基本信息

批准号：
8431376
负责人：
SCOTT W WALSH
金额：
$ 34.52万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-12-05 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8431376
关键词：
Abdomen Accounting Angiotensin II Antibodies Antioxidants Arteries Blood Pressure Blood Vessels Candidate Disease Gene Cardiovascular Diseases Cardiovascular Physiology Cell Adhesion Molecules Cell Culture Techniques Cells Cesarean section Chemistry Chronic Coculture Techniques DNA Methylation DNA Sequence Analysis Endothelium Enzyme-Linked Immunosorbent Assay Enzymes Epigenetic Process Extracellular Matrix Fatty acid glycerol esters Functional disorder Future Genes Health Human Immunohistochemistry In Situ Inflammation Inflammatory Inflammatory Response Intercellular adhesion molecule 1 Interleukin-8 Interstitial Collagenase Life Long-Term Effects Measures Mediating Methylation Modeling Molecular Morbidity - disease rate Neutrophil Activation Neutrophil Collagenase Neutrophil Infiltration Norepinephrine Nuclear Omentum Operative Surgical Procedures Oxidative Stress Pathology Patients Pattern Peroxidases Phenotype Phosphorylation Phosphotransferases Plasma Platelet Activation Poison Pre-Eclampsia Pregnancy Production Reactive Oxygen Species Recurrence Reporter Research Risk Role Smooth Muscle Myocytes System TNF gene Testing Thromboxane Production Thromboxanes Time Tissue Stains Tissues Transfection Tumor Necrosis Factor-alpha Two-Dimensional Gel Electrophoresis Vascular Smooth Muscle Vascular System Vasoconstrictor Agents Western Blotting Woman bisulfite cardiovascular disorder risk chemokine cyclooxygenase 2 cytokine fetal imprint inflammatory marker inhibitor/antagonist mortality mutant myosin phosphatase neutralizing antibody neutrophil novel oxidant stress pregnant programs public health relevance response subcutaneous vascular inflammation vasoconstriction

项目摘要

DESCRIPTION (provided by applicant): The over-arching hypothesis to be tested in this research is that vascular inflammation due to neutrophil infiltration is critical to the pathophysiology of preeclampsia. Neutrophils infiltrating the vasculature could cause inflammation and vasoconstriction by release of toxic compounds, such as reactive oxygen species (ROS), TNF??and thromboxane. In Specific Aim 1, we will determine if maternal vascular infiltration by neutrophils is a hallmark of preeclampsia, and associated with vascular inflammation. Subcutaneous fat, omental fat and placental tissue from pregnant patients will be collected and evaluated by immunohisto chemistry to assess vascular phenotypes of normal nonpregnant, normal pregnant and preeclamptic women, and to determine if neutrophil infiltration is restricted to the maternal compartment. We will stain tissues for CD66b (a neutrophil marker), intercellular adhesion molecule-1 (an endothelial adhesion molecule for neutrophils), interleukin-8 (IL-8, a potent neutrophil chemokine), NF-?B and COX-2 (hallmarks of inflammation), myeloperoxidase (a potent oxidizing enzyme), and matrix metalloproteinases 1 and 8. The in situ findings of vascular pathology observed in this Aim will drive the mechanistic studies of Aims 2-4 which will examine mechanisms for neutrophil induced vascular inflammation and vasoconstriction. In Specific Aim 2, we will evaluate whether neutrophils that have infiltrated into the intimal space could be responsible for transmitting inflammation to the vascular smooth muscle by co-culturing neutrophils with human vascular smooth cells. We will use transfection and cell culture to determine if neutrophils can activate NF-?B and induce expression of COX-2, IL-8 and thromboxane in human vascular smooth muscle cells (VSMC). In Specific Aim 3, we will use a myograph system with isolated omental vessels and cell culture of VSMC to examine cellular mechanisms whereby neutrophils could cause vasoconstriction and/or enhance vessel reactivity to vasoconstrictors. In Specific Aim 4, we will determine if neutrophils or neutrophil products induce epigenetic changes in vascular smooth muscle genes related to inflammation, and thus, may program adverse long-term effects on maternal cardiovascular function. Methylation patterns of candidate genes will be evaluated in vessels from women with normal pregnancies and women with preeclampsia by bisulfite DNA sequence analysis and COBRA (Combined Bisulfite Restriction Analysis). We will also determine if neutrophils or neutrophil products (ROS, TNF?) alter the methylation status of these genes in a similar pattern in cultured VSMC. This novel hypothesis may explain future health issues, including recurrent preeclampsia and future risk of cardiovascular disease.

描述（由申请人提供）：本研究要验证的首要假设是中性粒细胞浸润引起的血管炎症对子痫前期的病理生理至关重要。浸润血管的中性粒细胞可通过释放有毒化合物，如活性氧（ROS）、肿瘤坏死因子（TNF）等，引起炎症和血管收缩。凝血恶烷。在特异性目的1中，我们将确定中性粒细胞浸润母体血管是否是子痫前期的标志，并与血管炎症有关。收集孕妇皮下脂肪、大网膜脂肪和胎盘组织，通过免疫组化评估正常非妊娠、正常妊娠和子痫前期妇女的血管表型，并确定中性粒细胞浸润是否局限于母体腔室。我们将对组织进行CD66b（一种中性粒细胞标记物）、细胞间粘附分子-1（一种中性粒细胞内皮粘附分子）、白细胞介素-8 （IL-8，一种有效的中性粒细胞趋化因子）、NF-？B和COX-2（炎症的标志），髓过氧化物酶（一种强效氧化酶）和基质金属蛋白酶1和8。本研究中观察到的血管病理原位发现将推动Aims 2-4的机制研究，该研究将研究中性粒细胞诱导的血管炎症和血管收缩的机制。在Specific Aim 2中，我们将通过与人血管平滑肌细胞共培养中性粒细胞，评估浸润到内膜间隙的中性粒细胞是否负责将炎症传递到血管平滑肌。我们将使用转染和细胞培养来确定中性粒细胞是否可以激活NF-？B并诱导COX-2、IL-8和血栓素在人血管平滑肌细胞（VSMC）中的表达。在具体目标3中，我们将使用肌图系统与分离的大网膜血管和VSMC细胞培养来检查中性粒细胞引起血管收缩和/或增强血管对血管收缩剂的反应性的细胞机制。在Specific Aim 4中，我们将确定中性粒细胞或中性粒细胞产物是否会诱导与炎症相关的血管平滑肌基因的表观遗传变化，从而可能对母体心血管功能产生不良的长期影响。候选基因的甲基化模式将通过亚硫酸氢盐DNA序列分析和COBRA（联合亚硫酸氢盐限制分析）来评估正常妊娠妇女和子痫前期妇女血管中的候选基因。我们还将确定中性粒细胞或中性粒细胞产物（ROS， TNF?）是否在培养的VSMC中以类似的模式改变这些基因的甲基化状态。这一新的假说可以解释未来的健康问题，包括复发性子痫前期和未来心血管疾病的风险。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Matrix Metalloprotease-1 and Elastase Are Novel Uterotonic Agents Acting Through Protease-Activated Receptor 1.

Matrix Metalloprotease-1 和弹性蛋白酶是通过蛋白酶激活受体 1 发挥作用的新型子宫收缩剂。

DOI：
10.1177/1933719117732162
发表时间：
2018
期刊：
Reproductive sciences (Thousand Oaks, Calif.)
影响因子：
0
作者：
Walsh,ScottW;Nugent,WilliamH;Solotskaya,AnnaV;Anderson,CharlesD;Grider,JohnR;Strauss3rd,JeromeF
通讯作者：
Strauss3rd,JeromeF

Plasma from preeclamptic women stimulates transendothelial migration of neutrophils.