Daniel Baugh Institute for Functional Genomics and Computational Biology, Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA, 19107, USA.
J Neuroinflammation. 2012 May 24;9:97. doi: 10.1186/1742-2094-9-97.
Chronic alcohol use changes the brain's inflammatory state. However, there is little work examining the progression of the cytokine response during alcohol withdrawal, a period of profound autonomic and emotional upset. This study examines the inflammatory response in the central nucleus of the amygdala (CeA) and dorsal vagal complex (DVC), brain regions neuroanatomically associated with affective and cardiorespiratory regulation in an in vivo rat model of withdrawal following a single chronic exposure.
For qRT-PCR studies, we measured the expression of TNF-α, NOS-2, Ccl2 (MCP-1), MHC II invariant chain CD74, and the TNF receptor Tnfrsf1a in CeA and DVC samples from adult male rats exposed to a liquid alcohol diet for thirty-five days and in similarly treated animals at four hours and forty-eight hours following alcohol withdrawal. ANOVA was used to identify statistically significant treatment effects. Immunohistochemistry (IHC) and confocal microscopy were performed in a second set of animals during chronic alcohol exposure and subsequent 48-hour withdrawal.
Following a chronic alcohol exposure, withdrawal resulted in a statistically significant increase in the expression of mRNAs specific for innate immune markers Ccl2, TNF-α, NOS-2, Tnfrsf1a, and CD74. This response was present in both the CeA and DVC and most prominent at 48 hours. Confocal IHC of samples taken 48 hours into withdrawal demonstrate the presence of TNF-α staining surrounding cells expressing the neural marker NeuN and endothelial cells colabeled with ICAM-1 (CD54) and RECA-1, markers associated with an inflammatory response. Again, findings were consistent in both brain regions.
This study demonstrates the rapid induction of Ccl2, TNF-α, NOS-2, Tnfrsf1a and CD74 expression during alcohol withdrawal in both the CeA and DVC. IHC dual labeling showed an increase in TNF-α surrounding neurons and ICAM-1 on vascular endothelial cells 48 hours into withdrawal, confirming the inflammatory response at the protein level. These findings suggest that an abrupt cessation of alcohol intake leads to an acute central nervous system (CNS) inflammatory response in these regions that regulate autonomic and emotional state.
慢性酒精使用会改变大脑的炎症状态。然而，在酒精戒断期间，细胞因子反应的进展研究甚少，这是一个自主神经和情绪失调的深刻时期。本研究在体内大鼠模型中检查了中央杏仁核（CeA）和背侧迷走神经复合体（DVC）的炎症反应，这些脑区在神经解剖学上与情感和心肺调节有关，在单次慢性暴露后戒断。
对于 qRT-PCR 研究，我们测量了 TNF-α、NOS-2、Ccl2（MCP-1）、MHC II 不变链 CD74 和 TNF 受体 Tnfrsf1a 在 CeA 和 DVC 样本中的表达，这些样本来自接受液体酒精饮食 35 天的成年雄性大鼠，以及在酒精戒断后 4 小时和 48 小时接受类似治疗的动物。使用 ANOVA 确定具有统计学意义的治疗效果。在第二组动物中进行了慢性酒精暴露和随后的 48 小时戒断期间的免疫组织化学（IHC）和共聚焦显微镜检查。
在慢性酒精暴露后，戒断导致特定于先天免疫标记物 Ccl2、TNF-α、NOS-2、Tnfrsf1a 和 CD74 的 mRNA 表达显著增加。这种反应存在于 CeA 和 DVC 中，在 48 小时时最为明显。在戒断 48 小时时取出的样本的共聚焦 IHC 显示 TNF-α 染色周围存在表达神经标记物 NeuN 的细胞和与 ICAM-1（CD54）和 RECA-1 共标记的内皮细胞，这些标记物与炎症反应有关。同样，这两种大脑区域的结果都是一致的。
本研究表明，在 CeA 和 DVC 中，酒精戒断期间 Ccl2、TNF-α、NOS-2、Tnfrsf1a 和 CD74 的表达迅速诱导。48 小时时，IHC 双重标记显示 TNF-α 围绕神经元和血管内皮细胞上的 ICAM-1 增加，证实了蛋白质水平的炎症反应。这些发现表明，突然停止饮酒会导致这些调节自主神经和情绪状态的区域的中枢神经系统（CNS）急性炎症反应。
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