LIPID PEROXIDATION\CELL SIGNALING-CC14-INDUCED FIBROSIS

脂质过氧化细胞信号传导-CC14-诱导的纤维化

基本信息

批准号：
6524808
负责人：
DENNIS PETERSEN
金额：
$ 37.54万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-09-15 至 2005-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6524808
关键词：
AP1 protein aldehydes biological signal transduction cell death collagen environmental toxicology enzyme activity fibrosis genetic regulation genetic transcription laboratory rat lipid metabolism liver disorder metalloendopeptidases nuclear factor kappa beta peroxidation tissue /cell culture tissue inhibitor of metalloproteinases

项目摘要

DESCRIPTION: (Adapted from the Investigator's Abstract): The long-range goal of this project is to establish the mechanistic role(s) of aldehydic products of lipid peroxidation (4-hydroxynonenal [4-HNE] and malondialdehyde [MDA]) in chemical-induced liver fibrosis. It has become increasingly clear that these chemically reactive aldehydes display a spectrum of cellular effects ranging from enzyme inhibition to increasing transcription of specific genes involved in collagen synthesis and fibrosis. The investigator's working hypothesis predicts that AP-1 and NFkappaB are critical systems in Ito cells affected by 4-HNE and MDA the result(s) of which are manifested in an imbalance of collagen synthesis/degradation and dysregulation of hepatocyte death processes. This hypothesis will be systematically evaluated in 3 specific aims using hepatic nonparenchymal and parenchymal cells isolated form rat during the progression of CCl4-induced cirrhosis. In Specific Aim 1, the PI will establish the fibrogenic potential of 4-HNE and MDA as regulatory elements in AP-1 transcriptional dysregulation involving alpha1-(1) procollagen, metalloproteinase-1 (MMP-1) and tissue metalloproteinase inhibitor-1 (TIMP-1) genes in cultured Ito cells. Experiments proposed for Specific Aim 2 will investigate changes NFkB signaling pathways in Ito cells during the time course of CC14-induced hepatic fibrosis and determine how regulation of this pathway in Ito cells affects cell death in hepatocytes. In Specific Aim 3, experiments using co-cultures of hepatocytes, Kupffer cells and Ito cells will be performed to identify the cellular origin of 4-HNE or MDA and determine the ability of these aldehydes to diffuse into and effect adjacent cells. The effects of these aldehydes on Ito cells will be evaluated in terms of interactions with Kupffer cell-derived cytokines TNFalpha, and TGFbeta with the goal of identifying cellular sources and targets of cellular mediators that potentially influence pathways of cell death. These studies will greatly enhance the understanding of the mechanistic role(s) of aldehydic products of lipid peroxidation in chemical-induced liver injury and fibrosis. Identification of novel cellular targets and specific mechanisms could provide information leading to development of effective therapeutic interventions for ameliorating specific components of hepatic inflammatory and fibrotic processes.

描述：（改编自研究者摘要）：长期目标该项目旨在确定醛类产品的机械作用脂质过氧化（4-羟基壬烯醛 [4-HNE] 和丙二醛 [MDA]）化学物质引起的肝纤维化。越来越明显的是，这些具有化学活性的醛具有一系列细胞效应：从酶抑制到增加相关特定基因的转录胶原蛋白合成和纤维化。研究者的工作假设预测 AP-1 和 NFkappaB 是受 Ito 细胞影响的关键系统 4-HNE 和 MDA 其结果表现为胶原蛋白不平衡肝细胞死亡过程的合成/降解和失调。这将使用肝脏在 3 个特定目标中系统地评估假设在进展过程中从大鼠中分离出非实质细胞和实质细胞 CCl4 诱发的肝硬化。在具体目标 1 中，PI 将建立 4-HNE 和 MDA 作为 AP-1 调节元件的纤维化潜力涉及 alpha1-(1) 前胶原的转录失调，金属蛋白酶-1 (MMP-1) 和组织金属蛋白酶抑制剂-1 (TIMP-1) 培养的伊藤细胞中的基因。针对具体目标 2 提出的实验将研究时间过程中 Ito 细胞 NFkB 信号通路的变化 CC14 诱导的肝纤维化并确定该途径的调节方式在伊藤细胞中影响肝细胞的细胞死亡。在具体目标 3 中，实验将使用肝细胞、Kupffer 细胞和 Ito 细胞的共培养进行鉴定 4-HNE 或 MDA 的细胞起源并确定这些醛扩散到并影响邻近的细胞。这些的影响将根据与 Kupffer 的相互作用来评估 Ito 细胞上的醛细胞源性细胞因子 TNFα 和 TGFβ，目的是识别潜在影响细胞介质的细胞来源和目标细胞死亡的途径。这些研究将极大地增强对机制作用的理解化学性肝损伤中脂质过氧化醛产物的研究和纤维化。识别新的细胞靶点和具体机制可以提供有助于开发有效治疗方法的信息改善肝脏炎症特定成分的干预措施纤维化过程。