ALCOHOL-INDUCED MODULATION OF CYTOKINES & NITRIC OXIDE

酗酒增加患者对肺炎和结核病的易感性 是一种未知的机制造成的。乙醇可能通过以下途径抑制肺宿主防御 下调肿瘤坏死因子(α)和一氧化氮合成 肺泡巨噬细胞(AM)和募集中性粒细胞(PMN)。此外， 乙醇可增加吞噬细胞内铁(FeI)，而FeI可降低 免费杀灭细菌否。为了验证这一假设，我们将使用病毒 自由SPF大鼠(n-5/GP)，非酒精等热量饮食(NAI)，NAI 给予标准酒精饮食中剂量的乙醇(AR)，NAI，共6周 (慢性)和慢性给药剂量(慢性AR)。我们规定 四个具体目标：(A)。确定乙醇是否介导了下调 PM和AM中一氧化氮合酶II的表达减弱肺宿主防御。计划中的步骤 包括：将肺炎克雷伯菌或内毒素(LPS)直接注入大鼠体内 阿龙。固定时间外周血灌洗循环获取中性粒细胞和AM 中性粒细胞和AM释放游离NO变化的组织学研究 PMN和AM表面微电极的测量量测 ~(14)C-L精氨酸转化为瓜氨酸和NO的NOS II活性。 竞争反转法测定中性粒细胞和AM中一氧化氮合酶II的mRNA 转录酶聚合酶链式反应。确定是否下调监管 NO的1)与细菌杀灭减少有关，2)通过 乙醇或乙醛或3)细胞因子介导的细胞变化 发信号。(B)确定乙醇介导的肿瘤坏死因子抑制和 皮质酮水平升高下调中性粒细胞和AM中一氧化氮合酶II我们生病了 上调和下调的肿瘤坏死因子和皮质酮的作用 乙醇介导的一氧化氮合酶II、细菌杀灭和肺组织的改变 清除感染的能力(见a)。(C)决定乙醇是否- 直接作用于基因导致一氧化氮合酶II的介导下调 NOS II或NOS II活性的表达。我们将上下调节 腺苷、cAMP和Ca~(2+)对乙醇或乙醛的影响 介导的超氧化物歧化抑制(铁细胞色素-C-还原酶试验)， NO II和细菌杀灭(见a)。(D)决定乙醇是否- 介导的吞噬细胞FeI的增加或生物素的减少可减弱 NO的杀菌活性。我们将上下规范费和 生物素，测量它们在PMN和AM中的浓度，并评估它们的 对细菌杀灭、一氧化氮和一氧化氮合酶II及肺功能的影响 清除感染(见a)。这个项目将产生新的方法 用于预防/治疗酗酒者的机会性感染。

Alcoholism increases patient susceptibility to pneumonia and tuberculosis by an unknown mechanism. Ethanol may inhibit pulmonary host defense by down-regulating tumor necrosis factor(alpha) (TNFalpha) and NO synthesis in alveolar macrophage (AM) and recruited neutrophils (PMN). Moreover, ETOH increases phagocyte intracellular iron (Fei) which can decrease bacterial killing by free No. To test this hypothesis we will use virus free SPF rats (n-5/gp) on a non-alcoholic, isocaloric diet (NAI), NAI given a bolus dose of ETOH (AR), NAI on a standard alcohol diet for 6 wk (Chronic) and Chronic given a bolus dose of (Chronic AR). We stipulate four Specific Aims: (a). Determine whether ETOH-mediated down regulation of NOS II in PM and AM attenuates pulmonary host defense. Planned steps include: give rats K. pneumonia or endotoxin (LPS) directly into the lung. Obtain PMN and AM by lavage and circulating PMN from blood at fixed histology in relation to changes in PMN and AM release of free NO measured with microelectrodes at the surface of the PMN and AM. Measure NOS II activity by conversion of 14 C-L-arginine to citrulline and NO. Measure mRNA for NOS II in PMN and AM using competitive reverse transcriptase polymerase chain reaction. Determine if down-regulation of NO is 1) associated with decreased bacterial killing, 2) mediated by ETOH or acetaldehyde or 3) results from changes in cytokine-mediated cell signalling. (b) Determine whether ETOH-mediated suppression of TNF and increases of corticosterone down regulates NOS II in PMN and AM. We ill up and down regulated TNF and corticosterone assessing their effects on ETOH-mediated alterations in NOS II, bacterial killing and the lung's ability to clear an infection (see a). (c) Determine whether ETOH- mediated down regulation of NOS II results from a direct effect on gene expression for NOS II or NOS II activity. We will up and down regulated adenosine, cAMP and Ca2+ assessing their effect on ETOH or acetaldehyde- mediated inhibition of superoxide (ferricytochrome-c-reductase assay), NOS II and bacterial killing (see a). (d) Determine whether ETOH- mediated increases in phagocyte Fei or decreases of biotin attenuate the bactericidal activity of NO. We will up and down regulate Fei and biotin, measure their concentrations in PMN and AM and assess their effects on bacterial killing, No and NOS II and the lung's ability to clear an infection (see a). This project will engender new approaches for preventing/treating opportunistic infections in alcoholics.