Collagenolysis and Elastolysis in Urinary Incontinence

尿失禁中的胶原蛋白溶解和弹性组织溶解

• 批准号: 6934518
• 负责人: BERTHA CHEN
• 金额: $ 31.74万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6934518
• 关键词: alpha 1 antitrypsin; biopsy; clinical research; collagen; elastases; elastin; enzyme activity; estradiol; fibroblasts; gene expression; hormone regulation /control mechanism; human subject; immunocytochemistry; menstrual cycle; metalloendopeptidases; microarray technology; molecular pathology; polymerase chain reaction; postmenopause; progesterone; proteolysis; relaxin; tissue /cell culture; tissue inhibitor of metalloproteinases; transforming growth factors; urinary incontinence; vagina; western blottings; women' s health

DESCRIPTION (provided by applicant): Urinary incontinence is a major health problem in the United States, resulting in an 11% lifetime risk of surgery for women with incontinence and/or prolapse. The molecular pathophysiology is poorly understood, inhibiting the development of medical prophylaxis or therapy for this growing area of disability. Collagen and elastin are major supporting elements of pelvic structures. Thus, expression of collagenases, elastases and their inhibitors controls proteolysis of the extracellular matrix and may negatively impact urinary function. We hypothesize that stress urinary incontinence (SUI) results from abnormal pelvic collagen and elastin metabolism as a function of differential expression of the matrix metalloproteinases (MMPs) and elastases and their respective protein inhibitors, the tissue inhibitors of metalloproteinases (TIMPs) and A-1 anti-trypsin. First, we propose to investigate expression of both the proteolytic enzymes and their inhibitors in vaginal wall tissue isolated from women with SUI and continent women during the follicular and luteal phases of the menstrual cycle using quantitative-competitive reverse transcription polymerase chain reaction (QC-RT-PCR), Western blot, immunohistochemical analysis, and zymography. Second, we will compare the expression of MMPs and TIMPs as well as neutrophil elastase and A-1 anti-trypsin in vaginal tissue samples from continent and incontinent postmenopausal women to determine the effect of estrogen deprivation on RNA and protein expression as well as on collagen and elastin proteolytic activity. In addition, we will compare broad patterns of gene expression between continent and incontinent women using microarray analysis. Third, to assess the role of growth factors in the etiology of urinary dysfunction, we will examine relaxin and transforming growth factor-B (TGF-B) modulation of both collagenolysis and elastolysis in cultured fibroblasts derived from premenopausal continent and incontinent women. Because urinary dysfunction develops as women age, our fourth goal is to examine the ability of 17-B estradiol and progesterone to alter in vitro expression of the ratio of MMPs/TIMPs and total elastase activity/A-1 anti-trypsin in vaginal fibroblast cultures. Two-thirds of the burden of urinary incontinence is borne by women with prevalence rates of 14-41%. Our goal is to identify specific pathophysiologic changes related to reproductive events and aging which underlie the development of SUI as a first step to identifying potential targets for therapeutic intervention.

描述(申请人提供)：尿失禁在美国是一个主要的健康问题，导致大小便失禁和/或脱垂的女性一生中有11%的手术风险。分子病理生理学知之甚少，阻碍了对这一日益增长的残疾领域的医学预防或治疗的发展。胶原蛋白和弹性蛋白是骨盆结构的主要支撑元素。因此，胶原酶、弹性蛋白酶及其抑制物的表达控制细胞外基质的蛋白分解，并可能对排尿功能产生负面影响。我们假设压力性尿失禁(SUI)是由于盆腔胶原和弹性蛋白代谢异常所致，这是由于基质金属蛋白酶(MMPs)和弹性蛋白酶及其各自的蛋白抑制物、金属蛋白酶组织抑制物(TIMPs)和A-1抗胰蛋白酶的差异表达所致。首先，我们建议利用定量-竞争性逆转录聚合酶链式反应(QC-RT-PCR)、Western印迹、免疫组织化学分析和酶谱技术，研究SUI患者和正常女性月经周期卵泡期和黄体期阴道壁组织中蛋白水解酶及其抑制物的表达。其次，我们将比较绝经后失禁和失禁妇女阴道组织中MMPs和TIMPs以及中性粒细胞弹性蛋白酶和A-1抗胰蛋白酶的表达，以确定雌激素剥夺对RNA和蛋白质表达以及胶原和弹性蛋白分解活性的影响。此外，我们将使用微阵列分析比较大小便失禁和大小便失禁女性之间基因表达的广泛模式。第三，为了评估生长因子在尿失禁病因中的作用，我们将检测松弛素和转化生长因子-B(TGF-B)对来自绝经前失禁和失禁妇女的培养成纤维细胞的胶原酶分解和弹性分解的调节。由于尿路功能障碍随着女性年龄的增长而发展，我们的第四个目标是检测17-B雌二醇和孕酮在体外改变阴道成纤维细胞培养中MMPs/TIMPs和总弹力酶活性/A-1抗胰蛋白酶的比率的能力。三分之二的尿失禁负担是由患病率为14-41%的妇女承担的。我们的目标是确定与生殖事件和衰老相关的特定病理生理变化，这些变化是SUI发生的基础，作为确定潜在治疗干预靶点的第一步。