权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

An in vitro system for the study of epithelial bicarbonate secretion

用于研究上皮碳酸氢盐分泌的体外系统

基本信息

批准号：
7743851
负责人：
Jonathan D. Kaunitz
金额：
$ 18.08万
依托单位：
BRENTWOOD BIOMEDICAL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-01 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7743851
关键词：
Acids Address Animals Anions Architecture Bicarbonates Biological Models Caco-2 Cells Cell Culture Techniques Cell Line Cell model Cells Chloride Ion Chlorides Cultured Cells Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Diarrhea Disease Duodenum Epithelial Epithelial Cells Epithelium Female Fibrinogen Functional disorder Heterogeneity Impairment In Vitro Intestinal Obstruction Intestines Kidney Knowledge Laboratory Animals Lead Life Liver Lung Lung diseases Male Genital Organs Male Infertility Measurable Measurement Methodology Methods Modeling Molecular Organ Pancreas Pancreatic duct Pancreatitis Pathogenesis Pathology Perfusion Pharmaceutical Preparations Physiological Play Procedures Proteins Renal tubule structure Research Personnel Role Salivary Glands Sodium Bicarbonate Stomach Structure of choroid plexus System Testing Time Tissue Viability Tissues Villous absorption animal tissue apical membrane base gastrointestinal in vitro Model male monolayer public health relevance ratiometric reproductive solute tool

项目摘要

DESCRIPTION (provided by applicant): Bicarbonate secretion plays an essential role in the physiological functioning of the duodenum, airway, pancreas, liver, male reproductive system, and kidney. Impairment of bicarbonate secretion, such as is manifest in the disease cystic fibrosis, can produce severe pathology in some or all of these organs. The mechanism of bicarbonate secretion is still not fully understood, despite decades of intensive study. Part of the problem is the methodology inherent in its study, which can involve intestinal, pancreatic duct, and renal tubule perfusion, and indirect methods, usually in live animals or in intact tissue. All of these methods suffer from excessive expense, cumbersome and time-consuming procedures, the use of laboratory animals which entails considerable expense and near- prohibitive regulatory requirements, and heterogeneity and lack of viability of the tissues studied. A bicarbonate-secreting in vitro system, although overcoming most of these obstacles, has not yet been developed. We propose to develop a bicarbonate secreting system based on commonly studied epithelial cell lines that have attributes similar to bicarbonate secreting intact epithelia and are thus likely to have measurable bicarbonate secretion. Our methodology includes culturing Caco-2 intestinal cells, or alternatively MDCK or other epithelial-derived cultured cells, on semi-permeable supports that can then be mounted in a perfusion chamber that enables the measurement of bicarbonate secretion, in addition to ratiometric measurement of intracellular pH, and short-circuit current across the monolayer. In this fashion, we plan to generate data that will enable us to test several key hypotheses. PUBLIC HEALTH RELEVANCE: Bicarbonate (baking soda) is secreted by the intestine in order to neutralize acid that is secreted by the stomach. Bicarbonate is also secreted by other important organs such as the pancreas, liver, lung, and male reproductive system. Disorders of bicarbonate secretion, as are found in the disease cystic fibrosis, can cause severe problems such as lung disease, pancreatitis, intestinal obstruction, and male infertility. Study of the mechanism of bicarbonate secretion has been hampered by the lack of a suitable model other than the use of live animals. We propose a cell-culture-based system suitable for the study of bicarbonate secretion that will help us and other investigators study how bicarbonate is secreted. With this knowledge, we hope to further understand why problems with bicarbonate secretion can lead to such sever diseases such as cystic fibrosis.

描述(申请人提供)：碳酸氢盐分泌物在十二指肠、呼吸道、胰腺、肝脏、男性生殖系统和肾脏的生理功能中起着重要作用。碳酸氢盐分泌的障碍，如囊性纤维化，可在这些器官的部分或全部产生严重的病理。尽管经过了几十年的深入研究，但对碳酸氢盐的分泌机制仍不完全清楚。部分问题是其研究中固有的方法，可能涉及肠道、胰管和肾小管灌注，以及间接方法，通常是在活体动物或完整组织中。所有这些方法都面临着昂贵、繁琐和耗时的程序，使用实验室动物需要相当大的费用和近乎令人望而却步的监管要求，以及所研究的组织的异质性和缺乏活力。虽然克服了这些障碍中的大部分，但体外分泌碳酸氢盐的系统还没有开发出来。我们建议在通常研究的上皮细胞系的基础上开发一种碳酸氢盐分泌系统，这些上皮细胞株具有类似于碳酸氢盐分泌的完整上皮细胞的属性，因此可能具有可测量的碳酸氢盐分泌。我们的方法包括将Caco-2肠道细胞或MDCK或其他上皮来源的培养细胞培养在半透性支架上，然后将其安装在灌注室中，以便能够测量碳酸氢盐的分泌，以及细胞内pH的比率测量和单层上的短路电流。通过这种方式，我们计划生成数据，使我们能够测试几个关键假设。与公共健康相关：小苏打(小苏打)是由肠道分泌的，目的是中和胃分泌的酸。碳酸氢盐也由其他重要器官分泌，如胰腺、肝脏、肺和男性生殖系统。在囊性纤维症中发现的碳酸氢盐分泌障碍会导致严重的问题，如肺部疾病、胰腺炎、肠梗阻和男性不育。除了使用活体动物外，由于缺乏合适的模型，对碳酸氢盐分泌机制的研究一直受到阻碍。我们提出了一种基于细胞培养的系统，适用于碳酸氢盐分泌的研究，这将有助于我们和其他研究人员研究碳酸氢盐是如何分泌的。有了这些知识，我们希望进一步了解为什么碳酸氢盐分泌问题会导致囊性纤维化等严重疾病。