Anti-Inflammatory effect of Cholinergic Pathway:Ischemia

胆碱能途径的抗炎作用：缺血

基本信息

批准号：
6838214
负责人：
ELAHE T CROCKETT
金额：
$ 14.95万
依托单位：
MICHIGAN STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-12-23 至 2007-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6838214
关键词：
Kupffer&apos s cell acetylcholine alanine transaminase cholinergic receptors cytokine enzyme linked immunosorbent assay histopathology immunocytochemistry inflammation laboratory mouse leukocyte activation /transformation liver ischemia /hypoxia liver pharmacology neutrophil polymerase chain reaction reperfusion western blottings

项目摘要

DESCRIPTION (provided by applicant): Ischemia- reperfusion (IR) injury occurs in a wide spectrum of clinically important disorders such as myocardial infarction, stroke, hemorrhagic shock, and organ transplantation. IR leads to an acute inflammatory tissue injury in which the inflammatory mediator cytokines play an important role. The Pl's lab has previously reported that inhibition of cytokine release from the macrophages significantly attenuated IR liver injury in mice (Mosher et al 2001). Recent studies have indicated that acetylcholine, the principal vagus nerve neurotransmitter, inhibited cytokine release by macrophages, through a novel physiological mechanism, termed the cholinergic antiinflammatory pathway (Tracey KJ, 2002).Thus, the objective of this proposal is to evaluate the effect of pharmacological stimulation of cholinergic anti-inflammatory pathway in liver injury following hepatic IR. The hypothesis is that the pharmacological stimulation of cholinergic pathway via acetylchofine receptors (AChRs) inhibits cytokine release, thereby inhibiting the development of inflammatory liver injury following IR. To test this hypothesis, the following specific aims are proposed: I. Determine the inhibitory effect of AChRs stimulation on liver injury in response to hepatic IR in mice. The liver injury will be evaluated by liver histopathology and changes of the plasma alanine transaminase (ALT) levels. II. Determine the effect of AChRs stimulation on production of inflammatory cytokines in mice subjected to hepatic IR; tissue mRNA expression and plasma levels of the cytokines will be measured. III. Determine the effect of AChRs stimulation on neutrophil infiltration into the ischemic liver of mice subjected to IR. IV. Determine if liver macrophages (i.e., Kupffer cells) are the major target of cytokine inhibition via AchR stimulation in hepatic IR. An ex vivo perfused liver model will be applied. An in vivo partial hepatic IR model in mice, with 90 min of ischemia followed by specific reperfusion periods, will be applied. Selective AChR agonists and antagonists will be used as the pharmacological stimulants to evaluate the role of anti-inflammatory cholinergic pathway in hepatic IR injury. The liver injury will be evaluated by monitoring changes in plasma ALT levels, neutrophil infiltration, and liver histopathology. The local (tissue) and systemic (plasma) cytokine levels will be measured using the Western and ELISA techniques, respectively. The ex vivo liver perfusion model will determine the role of Kupffer cells in cytokine production and inhibition by AChR agonists. The results of these studies will provide new insight into the role of the cholinergic anti-inflammatory pathway in hepatic IR injury. The generated knowledge, which may be applied to a wide range of other IR related diseases, will assist in the development of novel therapeutic approaches for inflammatory disease.

描述（由申请人提供）：缺血再灌注（IR）损伤发生在各种临床重要的疾病中，例如心肌梗塞，中风，出血性休克和器官移植。 IR导致急性炎症组织损伤，其中炎症介质细胞因子起着重要作用。 PL的实验室先前已经报道说，从巨噬细胞中抑制细胞因子释放，显着减弱了小鼠的IR肝损伤（Mosher等，2001）。最近的研究表明，主要的阴性神经神经递质乙酰胆碱通过一种新的生理机制抑制了巨噬细胞释放的细胞因子释放，称为胆碱能抗炎途径（Tracey KJ，2002）。肝ir。假设是，通过乙酰链蛋白受体（ACHR）对胆碱能途径的药理刺激抑制了细胞因子释放，从而抑制了IR后炎症性肝损伤的发展。为了检验该假设，提出了以下特定目的：I。确定小鼠肝脏刺激对肝脏损伤的抑制作用。肝损伤将通过肝组织病理学和血浆丙氨酸转氨酶（ALT）水平的变化进行评估。 ii。确定ACHRS刺激对经受肝IR的小鼠炎性细胞因子的产生的影响；将测量组织mRNA表达和血浆水平。 iii。确定ACHRS刺激对受IR的小鼠缺血性肝脏浸润的影响。 iv。确定肝巨噬细胞（即库普弗细胞）是否是通过肝IR中ACHR刺激的细胞因子抑制的主要靶标。将应用离体灌注肝模型。将应用小鼠的体内部分肝IR模型，缺血90分钟，然后使用特定的再灌注周期。选择性ACHR激动剂和拮抗剂将被用作药理兴奋剂，以评估抗炎胆碱能途径在肝IR损伤中的作用。肝损伤将通过监测血浆ALT水平，中性粒细胞浸润和肝组织病理学的变化来评估。将分别使用西方和ELISA技术测量局部（组织）和全身（血浆）细胞因子水平。离体肝灌注模型将确定库普弗细胞在细胞因子产生和ACHR激动剂抑制作用中的作用。这些研究的结果将为胆碱能抗炎途径在肝IR损伤中的作用提供新的见解。所产生的知识可以应用于多种其他相关疾病，将有助于开发新型的炎性疾病治疗方法。