NITRIC OXIDE AND INSULIN IN ISLET TRANSPLANTATION

胰岛移植中的一氧化氮和胰岛素

• 批准号: 6177536
• 负责人: CHARLES D MILLS
• 金额: $ 20.74万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-15 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6177536
• 关键词: RNase protection assay; autologous transplantation; blood glucose; cell type; diabetes mellitus; flow cytometry; gene targeting; genetically modified animals; homologous transplantation; hypoglycemia; insulin; laboratory mouse; laboratory rat; macrophage; nitric oxide; nitric oxide synthase; pancreatic islet transplantation; tissue /cell culture; transplant rejection; transplantation immunology

The goal of this investigation is to increase the success of clinical islet transplantation. Two interrelated hypotheses are put forth to achieve this goal. First, Nitric Oxide (NO) is postulated to be a prime mediator of islet dysfunction. Second, a previously unrecognized ability of insulin to decrease NO production is proposed to explain its islet-protective activity. Both of these hypotheses will be tested directly by focusing the experiments in this investigation on the use of inducible Nitric Oxide Synthase "knockout" mice (iNOS-/-). Specific Aim I will determine the role of macrophage NO in islet graft rejection. Based on preliminary and published results from this laboratory, NO is proposed to inhibit islets in 2 "waves": an early wave that occurs 1-2 days posttransplant of syngeneic or allogeneic islets is responsible for early islet dysfunction. A second wave NO-mediated islet dysfunction is proposed to occur during classical allograft rejection. The early wave will be investigated by comparing syngeneic islet function in diabetic iNOS-/- and iNOS-/+ mice; the second wave will be investigated using allogeneic islets in these hosts. Inhibitors of NO will then be used to determine how to best increase islet function. Preliminary and published evidence from this laboratory suggests that inhibiting NO in a clinically applicable way does increase the success of islet transplantation. Islets will be implanted in the intraperitoneal cavity in this investigation because intragraft NO production and allograft rejection responses can readily be followed at this site. Mice will be the primary species used because, like humans, they are "low" producers of NO; rats will be used in selected experiments because of their larger size. Specific Aim II will determine the mechanism by which insulin promotes islet function. A dominant paradigm has been that insulin allows transplanted beta-cells to "rest". However, preliminary evidence indicates that insulin administration to diabetic mice or rats decreases macrophage iNOS mRNA expression and NO production. Therefore, specific experiments will again compare iNOS- /- and iNOS-/+ mice to directly test the hypothesis that the islet- protective activity of insulin results from decreasing NO production. Finally, hyperglycemic clamps will be employed to maintain hyperglycemia in order to determine if the islet-protective activity of insulin depends on its ability to lower blood glucose. The results of this investigation should provide important new information that will increase the success of clinical islet transplantation as well as providing basic information on what role NO plays in rejection of cell transplants, and how NO production is regulated by insulin.

这项调查的目的是增加临床胰岛的成功 移植。 提出两个相互关联的假设以实现 这个目标。 首先，一氧化氮（NO）被认为是素数 胰岛功能障碍的介体。 第二，以前未被认可 胰岛素降低的能力不提出任何生产来解释其 胰岛保护活动。 这两个假设将进行测试 直接通过将实验集中在此调查中 诱导型一氧化氮合酶“基因敲除”小鼠（iNOS - / - ）。 具体的 目的我将确定巨噬细胞NO在胰岛移植中的作用 拒绝。 基于此的初步和发表的结果 实验室，提议在2个“波”中抑制胰岛：早期波 在同烯或同种异体胰岛移植后发生1-2天是 负责早期胰岛功能障碍。 第二波未介导的 提议在经典同种异体移植期间发生胰岛功能障碍 拒绝。 将通过比较同步性来研究早期浪潮 糖尿病iNOS - / - 和iNOS - /+小鼠中的胰岛功能；第二波 将使用这些宿主中的同种异体胰岛进行研究。 抑制剂 然后，将使用否来确定如何最好地增加胰岛功能。 该实验室的初步和已发表证据表明 以临床适用的方式抑制NO确实会增加 胰岛移植。 胰岛将植入腹膜内 这项调查的腔体是因为内部没有生产和 在此站点可以很容易地遵循同种异体移植拒绝反应。 老鼠 将是使用的主要物种，因为像人类一样，它们“低” No的生产者；大鼠将在选定的实验中使用 它们更大的尺寸。 具体目标II将通过 胰岛素促进胰岛功能。 主要的范式已经 该胰岛素允许移植的β细胞“休息”。 然而， 初步证据表明胰岛素给予糖尿病 小鼠或大鼠降低巨噬细胞mRNA的表达和无 生产。 因此，特定的实验将再次比较iNos- / - 和iNOS - /+小鼠直接检验胰岛的假设 胰岛素的保护活性导致没有生产降低。 最后，将采用高血糖夹来维持 高血糖以确定是否的胰岛保护活性是否 胰岛素取决于其降低血糖的能力。 结果 这项调查应提供重要的新信息 增加临床胰岛移植的成功并提供 关于在拒绝细胞中没有扮演的角色的基本信息 移植，以及如何通过胰岛素调节无生产。

项目成果

Insulin down-regulates the inducible nitric oxide synthase pathway: nitric oxide as cause and effect of diabetes?

• DOI: 10.4049/jimmunol.159.11.5329
• 发表时间: 1997-12
• 期刊: Journal of immunology
• 影响因子: 4.4
• 作者: R. Stevens;D. Sutherland;J. Ansite;M. Saxena;T. Rossini;B. Levay-Young;B. Hering;C. Mills

Augmentation of an antitumor CTL response In vivo by inhibition of suppressor macrophage nitric oxide.

• DOI: 10.4049/jimmunol.163.11.5877
• 发表时间: 1999-12
• 期刊: Journal of immunology
• 影响因子: 4.4
• 作者: Marianne Medot-Pirenne;M. Heilman;Malinee Saxena;Patricia E. McDermott;Charles D. Mills