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Thrombin Effects on Decidual TLR Expression

凝血酶对蜕膜 TLR 表达的影响

基本信息

批准号：
8378429
负责人：
Vikki M Abrahams
金额：
$ 21.21万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-01 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8378429
关键词：
Abruptio Placentae Affect Agonist Allogenic Bacterial Infections Birth Cell Culture Techniques Cells Complement Coupled Decidua Decidual Cell Dendritic Cells Disseminated Intravascular Coagulation Endometrial Endothelial Cells Event Gene Expression Generations Genes Genital system Gestational Age Health Heat shock proteins Hemorrhage Host Defense Human IRAK2 gene IRAK4 gene Immune Immune System Diseases Immune response Immune system Immunoassay Immunofluorescence Immunologic Immunohistochemistry Infection Infection of amniotic sac and membranes Inflammation Inflammatory Inflammatory Response Interleukin-8 Ligands Link Lipopolysaccharides Matrix Metalloproteinases Mediating Messenger RNA Microarray Analysis Microbe Microdissection Modeling Molecular Molecular Profiling Morbidity - disease rate Mus Mutant Strains Mice NF-kappa B Natural Immunity Natural Killer Cells Pathway interactions Patients Pattern Peptidoglycan Perinatal Placenta Poly I-C Predisposition Pregnancy Premature Birth Production Proteinase-Activated Receptors Public Health RNA Interference Reaction Receptor Gene Receptor Signaling Recurrence Regulation Reverse Transcriptase Polymerase Chain Reaction Role Signal Transduction Signaling Protein Stream Stromal Cells TLR2 gene TLR4 gene Testing Thrombin Time Tissues Toll-like receptors Western Blotting chemokine cytokine in vivo insight interest laser capture microdissection macrophage microbial mortality neutrophil pathogen pregnant preterm premature rupture of membranes protein expression receptor receptor expression receptor function research study response stress protein

项目摘要

Decidual hemorrhage/abruption results in intense local thrombin generation and is associated with both preterm premature rupture of the membrane (PPROM) and chorioamnionitis (CAM). Prior studies indicate that thrombin, acting via protease activated receptors (PARs), induces an intense decidual cytokine and proteolytic response. The link between abruption and CAM suggests an impaired immune response. Tolllike receptors (TLRs) initiate the host innate immune response. By promoting the innate immune response, TLRs provide the first line of defense against an array of microbial pathogens. We now demonstrate that decidual cells express TLRs and their intermediate signaling proteins. Importantly, we demonstrate that thrombin down-regulates decidual cell TLR expression and signaling. Finally, we demonstrate that abruption-associated PTD with or without CAM are accompanied by decreased decidual TLR expression. We postulate that decidual hemorrhage leads to intense local thrombin generation which paradoxically induces a local aseptic inflammatory reaction and promotes ascending genital tract infections by downregulating TLR expression and function. We propose four Specific Aims to assess the interactions between thrombin and TLRs expressed by all decidual cells. 1) Immunohistochemistry, immunofluorescence and microdissection coupled with quantitative RT-PCR will be utilized to quantify the association between altered TLR expression and abruption-associated PTD with and without related CAM. 2) To determine the mechanism(s) by which thrombin down-regulates decidual cell-expressed TLR levels and function. Studies will dissect out the role of PARs in the expression of TLRs and their downstream signaling intermediates as well as cytokine and NFkB expression. These studies will use agonists, antagonists and small interference RNA (siRNAs) in cell culture. 3) To determine the functional effects of thrombin on TLR-ligand interactions. Cultured decidual cells will be treated with thrombin vs. control and then exposed to TLR-2, 3 and 4 ligands. Endpoints of these studies will be assessed by microarray analysis, RT-PCR and immunoassays as well as assessment of the NFkB components by western blotting. 4) Lastly, in coordination with Projects I and III of this PO1, we will utilize a murine model to study the effects of thrombin on TLR expression and function as well as the susceptibility to bacterial infection. These studies will provide unique insights into the fundamental mechanisms underlying abruption associated PTD and dissect out the role of innate immune dysfunction in this major public health problem.

蜕膜出血/早剥导致强烈的局部凝血酶生成，并与两者相关