Determinants of sinusoidal endothelial cell phenotype

正弦内皮细胞表型的决定因素

• 批准号: 6936525
• 负责人: LAURIE D DELEVE
• 金额: $ 31.48万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-15 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6936525
• 关键词: alcoholic liver cirrhosis; cell cell interaction; fibroblast growth factor; fibrosis; growth factor; hepatocyte growth factor; hormone regulation /control mechanism; immunocytochemistry; laboratory rat; liver; liver cells; liver circulation; liver disorder; microarray technology; paracrine; phenotype; protein structure function; proteomics; vascular endothelial growth factors; vascular endothelium

DESCRIPTION (provided by applicant): Capillarization precedes fibrosis in alcoholic liver disease. In capillarization, sinusoidal endothelial cells (SEC) lose fenestrae and form a basement membrane, a loss of phenotype that has not been examined previously. We propose to examine determinants of SEC phenotype in a series of in vitro and in vivo experiments. In vitro, isolated cells will be examined in homotypic culture and in co-culture with other liver cells to determine whether SEC phenotype is regulated by paracrine regulation or by heterotypic contact. The in vitro studies will utilize cells isolated from normal rats and from liver disease models, notably models of steatosis, alcoholic liver disease and fibrosis using thioacetamide. Studies in SEC from liver disease models will examine changes in response to determinants of normal SEC phenotype by heterotypic cells. Studies in hepatocytes and stellate cells from liver disease models will look for changes in factors that determine SEC phenotype. A proteomic approach will be used to identify and examine proteins involved in paracrine regulation. In vivo studies will provide confirmation of the relevance of the soluble factors implicated as determinants of SEC phenotype using continuous intraportal infusions of the soluble factors or their inhibitors in normal rats or rats from the above mentioned liver disease models by monitoring changes in liver histology. Differences in gene expression between SEC from normal and liver disease model rats will be characterized to understand what regulates SEC phenotype. These studies will be used to identify markers of SEC that are altered in the models, to confirm the findings of the proteomic studies and to identify common pathways that regulate expression of genes that are expressed in normal SEC, but not in SEC isolated from liver disease models.

描述（由申请人提供）：酒精性肝病中毛细血管化先于纤维化。在毛细血管化过程中，窦内皮细胞 (SEC) 失去窗孔并形成基底膜，这是一种先前未检测到的表型损失。我们建议在一系列体外和体内实验中检查 SEC 表型的决定因素。在体外，分离的细胞将在同型培养物和与其他肝细胞共培养物中进行检查，以确定 SEC 表型是否受到旁分泌调节或异型接触的调节。体外研究将利用从正常大鼠和肝病模型中分离的细胞，特别是使用硫代乙酰胺的脂肪变性、酒精性肝病和纤维化模型。肝脏疾病模型的 SEC 研究将检查异型细胞对正常 SEC 表型决定因素的反应变化。对肝病模型的肝细胞和星状细胞的研究将寻找决定 SEC 表型的因素的变化。蛋白质组学方法将用于鉴定和检查参与旁分泌调节的蛋白质。体内研究将通过监测肝脏组织学的变化，在正常大鼠或来自上述肝病模型的大鼠中连续门静脉内输注可溶性因子或其抑制剂，以确认作为 SEC 表型决定因素的可溶性因子的相关性。将表征正常大鼠和肝病模型大鼠 SEC 之间基因表达的差异，以了解调节 SEC 表型的因素。这些研究将用于鉴定模型中发生改变的 SEC 标记，以证实蛋白质组学研究的结果，并确定调节在正常 SEC 中表达但在从肝病分离的 SEC 中不表达的基因表达的常见途径模型。