ALCOHOL-INDUCED MODULATION OF CYTOKINES & NITRIC OXIDE

酗酒增加病人对肺炎和肺结核的易感性 一种未知的机制。 乙醇可能通过以下方式抑制肺宿主防御： 下调肿瘤坏死因子（α）（TNF α）和NO合成 肺泡巨噬细胞（AM）和募集的中性粒细胞（PMN）。 此外，委员会认为， ETOH增加吞噬细胞胞内铁（Fei）， 为了验证这一假设，我们将使用病毒 游离SPF大鼠（n-5/gp）接受非酒精等热量饮食（NAI），NAI 给予ETOH（AR）、NAI和标准酒精饮食6周 （慢性）和慢性给予推注剂量的（慢性AR）。 我们规定 四个具体目标：（一）。确定ETOH介导的下调是否 PM和AM中NOS Ⅱ的表达减弱了肺宿主的防御功能。 计划采取的步骤 包括：给予大鼠K。肺炎或内毒素（LPS）直接进入 肺。灌洗法获得PMN和AM，并在固定时间从血液中循环PMN 组织学与PMN和AM释放NO变化关系 在PMN和AM的表面用微电极测量。 测量 14 C-L-精氨酸转化为瓜氨酸和NO的NOS II活性。 竞争性逆转录酶法检测PMN和AM中NOS Ⅱ mRNA的表达 转录酶聚合酶链反应。 确定是否下调 1）与减少的细菌杀伤有关，2）由 ETOH或乙醛，或3）由甜菜碱介导的细胞 信号装置. (b)确定ETOH介导的TNF抑制和 皮质酮升高可下调PMN和AM中NOS Ⅱ的表达。 我们会 上调和下调TNF和皮质酮，评估其对 ETOH介导的NOS II、细菌杀伤和肺组织的改变 清除感染的能力（见a）。(c)确定ETOH是否- NOSII介导的下调是由于基因表达的直接影响， NOS II表达或NOS II活性。 我们将上下调节 腺苷、cAMP和Ca 2+评估其对ETOH或乙醛的作用- 介导的超氧化物抑制（铁细胞色素-C-还原酶测定）， NOS II和细菌杀灭（见a）。 (d)确定ETOH是否- 介导的吞噬细胞Fei的增加或生物素的减少会减弱 杀菌活性NO。我们将上下调节费， 生物素，测量其在PMN和AM中的浓度，并评估其 对细菌杀灭、NO和NOS II以及肺的 清除感染（见a）。 该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.