ETHANOL-INDUCED ALTERATIONS OF MONOKINE PRODUCTION

单核细胞功能的改变是导致 酒精引起的免疫和代谢异常。 乙醇诱导 前列腺素E2（PGE 2）是许多酒精的介质 相关的M0和T淋巴细胞缺陷。 的产生减少 据报道，在大鼠中发现了炎症单核因子，例如TNFalpha， 急性乙醇摄取 我们对人类M0的数据表明， 乙醇处理降低了生物活性M0的产生 TNF α和IL-6的产生伴随着TNF α和IL-6的产生增加， 抑制性M0介质，如PGE 2和转化生长因子β （TGF β）。 高PGE 2水平下调Mo TNF α的产生， 环腺苷酸（cAMP）升高。 然而，我们的数据表明， 乙醇调节单核因子的PGE 2非依赖性途径。 乙醇 已显示增加淋巴细胞中的cAMP水平。 增加 M0 cAMP水平具有调节生物活性TNF α产生的潜力 和基因表达。 因此，我们的假设是，乙醇可以 调节单核因子的产生，特别是TNF α的产生， 通过直接增加M0 cAMP水平的cAMP依赖性途径。 此外，慢性乙醇治疗对M0的差异效应 TNF α的产生可能与酒精诱导的重复性PGE 2有关 升高可以使M0对PGE 2介导的下调 慢性酒精治疗期间TNF α的产生。 因此我们 建议研究cAMP参与急性心肌梗死的机制， 慢性乙醇处理改变了MO TNF α、IL-6的产生， 和TGF β在mRNA和生物活性水平上的表达。 变化 乙醇暴露后细胞内cAMP以及 cAMP激动剂和拮抗剂对乙醇诱导的 将评估单核因子的产生。 PGE 2在乙醇中的作用 还将研究诱导的单核因子和cAMP改变。 在 除了影响cAMP信号转导，乙醇还可以放大 通过Ca++动员进行信号转导。 细胞内Ca++水平 在大鼠M0中对乙醇的反应迅速增加。 CA++ 增加导致M0 PGE 2的产生，这有可能 调节单核因子的产生。 因此，我们假设， M0中乙醇相关的细胞内Ca++水平增加有助于 酒精治疗后M0反应的改变 所以我们提出 研究乙醇引起的细胞内Ca ~（++）水平的变化 并将它们与其对M0 PGE 2产生的影响相关联。 醇 将在个体M0中评价诱导的细胞内Ca++水平 在阿托氟上

Altered monocyte functions are major contributors to the development of alcohol induced immune and metabolic abnormalities. Ethanol-induced elevated M0 Prostaglandin E2 (PGE2) is the mediator of many alcohol related M0 and T lymphocyte defects. Decreased production of inflammatory monokines, such as TNFalpha, has been reported in rats after acute ethanol uptake. Our data on human M0 indicate that single in vitro ethanol treatment decreases the production of biologically active M0 TNFalpha and IL-6 with concomitant increase in the production of inhibitory M0 mediators, such as PGE2 and Transforming Growth Factorbeta (TGFbeta). High PGE2 levels downregulate Mo TNFalpha production via elevated cyclic AMP (cAMP). However our data indicate the existence of a PGE2-independent pathway for monokine regulation by ethanol. Ethanol has been shown to increase cAMP levels in lymphoid cells. Increases in M0 cAMP level has the potential to regulate bioactive TNFalpha production and gene expression. Therefore, our hypothesis is that ethanol can regulate the production of monokines, particularly that of TNFalpha, via a cAMP dependent pathway by directly increasing M0 cAMP levels. furthermore, the differential effect of chronic ethanol treatment on M0 TNFalpha production may be related to alcohol induced repetitive PGE2 elevations which can desensitize M0 to PGE2 mediated downregulation of TNFalpha production during chronic alcohol treatment. Consequently, we propose to study the involvement of cAMP in the mechanism by which acute and chronic ethanol treatment alters the production of MO TNFalpha, IL-6 and TGFbeta at the levels of mRNA and bioactivity. Changes in the intracellular cAMP following ethanol exposure as well as the effect of cAMP agonists and antagonists on the ethanol induced alterations of monokine production will be assessed. The role of PGE2 in ethanol induced monokine and cAMP alterations will also be investigated. In addition to affecting cAMP signal transduction, ethanol can also amplify signal transduction via Ca++ mobilization. Intracellular Ca++ levels were shown to rapidly increase in response to ethanol in rat M0. Ca++ increase results in M0 PGE2 production which has the potential for regulation of monokine production. Consequently, we hypothesize that ethanol related increases in intracellular Ca++ levels in M0 contribute to altered M0 responses after alcohol treatment. Therefore, we propose to study the ethanol induced changes in the level of intracellular Ca++ and correlate them with its effect on M0 PGE2 production. Alcohol induced intracellular Ca++ levels will be evaluated in the individual M0 on the Attofluor.