Mandrekar Pranoti, Dolganiuc Angela, Bellerose Gary, Kodys Karen, Romics Laszlo, Nizamani Rabia, Szabo Gyongyi
University of Massachusetts Medical School, Department of Medicine, Worcester 01605-2324, USA.
Alcohol Clin Exp Res. 2002 Nov;26(11):1609-14. doi: 10.1097/01.ALC.0000036926.46632.57.
Nuclear translocation and DNA binding of the nuclear factor kappaB (NF-kappaB) is an early event in inflammatory cell activation in response to stimulation with bacterial components or cytokines. Cell activation via different receptors culminates in a common pathway leading to NF-kappaB activation and proinflammatory cytokine induction. We have previously shown that acute alcohol inhibits NF-kappaB activation by lipopolysaccharide (LPS) in human monocytes. Here we investigated whether acute alcohol treatment of human monocytes also inhibits NF-kappaB when induced through activation of the interleukin (IL)-1 or tumor necrosis factor (TNF) receptors.
Human peripheral blood monocytes were treated with LPS, TNFalpha, and IL-1beta in the presence or absence of 25mM alcohol for 1 hr. NF-kappaB activation was determined by electrophoretic mobility shift assays using nuclear extracts. Inhibitory kappaB(alpha) (IkappaB(alpha)) was estimated by Western blotting in cytoplasmic extracts. Chinese hamster ovary cells expressing human CD14 were treated with LPS in the presence or absence of alcohol to study NF-kappaB and IkappaB(alpha) regulation.
Our results indicate that acute alcohol inhibits IL-1beta- and TNFalpha-induced NF-kappaB activation. We further show in CD14/toll-like receptor 4-expressing Chinese hamster ovary cells the specificity of alcohol-mediated inhibition of NF-kappaB via the toll-like receptor 4/CD14 receptors. Inhibition of NF-kappaB by acute alcohol was concomitant with decreased levels of the IkappaB(alpha) molecule in the cytoplasm of LPS, IL-1, and TNFalpha-activated monocytes.
These data suggest a unique, IkappaB(alpha)-independent pathway for the inhibition of NF-kappaB activation by acute alcohol in monocytes. Universal inhibition of NF-kappaB by acute alcohol via these various receptor systems suggests a target for the effects of alcohol in the NF-kappaB activation cascade that is downstream from IkappaB(alpha) degradation. Further, these results demonstrate that acute alcohol is a potent inhibitor of NF-kappaB activation by mediators of early (LPS) or late (IL-1, TNF(alpha)) stages of inflammation in monocytes.
核因子κB(NF-κB)的核转位及DNA结合是炎症细胞在受到细菌成分或细胞因子刺激后激活过程中的早期事件。通过不同受体激活细胞最终会汇聚到一条共同途径,导致NF-κB激活及促炎细胞因子的诱导。我们之前已经表明,急性酒精可抑制人单核细胞中脂多糖(LPS)诱导的NF-κB激活。在此,我们研究了急性酒精处理人单核细胞在通过白细胞介素(IL)-1或肿瘤坏死因子(TNF)受体激活而诱导NF-κB时是否也会产生抑制作用。
在存在或不存在25mM酒精的情况下,用人外周血单核细胞与LPS、TNFα和IL-1β一起处理1小时。使用核提取物通过电泳迁移率变动分析来测定NF-κB激活。通过蛋白质印迹法在细胞质提取物中估计抑制性κBα(IkappaBα)。在存在或不存在酒精的情况下,用LPS处理表达人CD14的中国仓鼠卵巢细胞,以研究NF-κB和IkappaBα的调节。
我们的结果表明,急性酒精可抑制IL-1β和TNFα诱导的NF-κB激活。我们进一步在表达CD14/ Toll样受体4的中国仓鼠卵巢细胞中表明,酒精通过Toll样受体4/CD14受体介导的对NF-κB的抑制具有特异性。急性酒精对NF-κB的抑制与LPS、IL-1和TNFα激活的单核细胞细胞质中IkappaBα分子水平的降低同时发生。
这些数据表明,急性酒精在单核细胞中抑制NF-κB激活存在一条独特的、不依赖IkappaBα的途径。急性酒精通过这些各种受体系统对NF-κB的普遍抑制表明,酒精在NF-κB激活级联反应中的作用靶点位于IkappaBα降解的下游。此外,这些结果表明,急性酒精是单核细胞中炎症早期(LPS)或晚期(IL-1、TNFα)介质激活NF-κB的有效抑制剂。