Increased expression of M1 and M2 phenotypic markers in isolated microglia after four-day binge alcohol exposure in male rats.
Increased expression of M1 and M2 phenotypic markers in isolated microglia after four-day binge alcohol exposure in male rats.
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DOI：
10.1016/j.alcohol.2017.02.175
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发表时间：
2017-08
期刊：
影响因子：
--
通讯作者：
Nixon K
中科院分区：
文献类型：
--
作者：
Peng H;Geil Nickell CR;Chen KY;McClain JA;Nixon K
小胶质细胞活化和神经炎症是神经退行性疾病的常见特征，包括酒精使用障碍（AUD）。当被激活时，小胶质细胞跨越一系列不同的表型，从经典激活的促炎（M1）小胶质细胞/巨噬细胞到交替激活的促生长（M2）小胶质细胞/巨噬细胞。识别小胶质细胞表型对于理解小胶质细胞在AUD发病机制中的作用至关重要。因此，雄性大鼠每8小时灌胃25%（w/v）乙醇或等热量对照饮食，持续4天，并在酒精暴露后0、2、4和7天处死（例如，T0、T2等）。采用Percoll密度梯度离心法从海马和内嗅皮质分离小胶质细胞。用小胶质细胞表面抗原标记细胞并通过流式细胞术分析。与先前的研究一致，分离的细胞产生了高度富集的脑巨噬细胞/小胶质细胞群（纯度>95%），通过巨噬细胞/小胶质细胞抗原CD 11b染色证明。通过M1表面标志物、主要组织相容性复合物（MHC）II、CD 32、CD 86和M2表面标志物、CD 206（甘露糖受体）的表达来评价CD 11b + CD 45低水平小胶质细胞的极化状态。乙醇处理动物开始显示T0时M1和M2标记物表达增加（p = n.s.），在T2时间点有显著变化。在T2，M1标记物、MHC-II、CD 86和CD 32在海马和内嗅皮质中的表达增加（p < 0.05），而M2标记物、CD 206仅在内嗅皮质中显著增加（p < 0.05）。到T4时，所有效应均恢复至对照水平。总之，4天的酗酒暴露产生了短暂的增加M1（MHC-II，CD 32，和CD 86）和M2（CD 206）的小胶质细胞从内嗅皮层和海马分离的人口。因此，这些发现，既促炎和潜在的有益的，促进恢复的小胶质细胞表型，可以观察到后，酒精的破坏性暴露是至关重要的，我们的小胶质细胞的作用的理解在AUD的发病机制。
Microglia activation and neuroinflammation are common features of neurodegenerative conditions, including alcohol use disorders (AUDs). When activated, microglia span a continuum of diverse phenotypes ranging from classically activated, pro-inflammatory (M1) microglia/macrophages to alternatively activated, growth-promoting (M2) microglia/macrophages. Identifying microglia phenotypes is critical for understanding the role of microglia in the pathogenesis of AUDs. Therefore, male rats were gavaged with 25% (w/v) ethanol or isocaloric control diet every 8 h for 4 days and sacrificed at 0, 2, 4, and 7 days after alcohol exposure (e.g., T0, T2, etc.). Microglia were isolated from hippocampus and entorhinal cortices by Percoll density gradient centrifugation. Cells were labeled with microglia surface antigens and analyzed by flow cytometry. Consistent with prior studies, isolated cells yielded a highly enriched population of brain macrophages/microglia (>95% pure), evidenced by staining for the macrophage/microglia antigen CD11b. Polarization states of CD11b+CD45low microglia were evaluated by expression of M1 surface markers, major histocompatibility complex (MHC) II, CD32, CD86, and M2 surface marker, CD206 (mannose receptor). Ethanol-treated animals begin to show increased expression of M1 and M2 markers at T0 (p = n.s.), with significant changes at the T2 time point. At T2, expression of M1 markers, MHC-II, CD86, and CD32 were increased (p < 0.05) in hippocampus and entorhinal cortices, while M2 marker, CD206, was increased significantly only in entorhinal cortices (p < 0.05). All effects resolved to control levels by T4. In summary, four-day binge alcohol exposure produces a transient increase in both M1 (MHC-II, CD32, and CD86) and M2 (CD206) populations of microglia isolated from the entorhinal cortex and hippocampus. Thus, these findings that both pro-inflammatory and potentially beneficial, recovery-promoting microglia phenotypes can be observed after a damaging exposure of alcohol are critically important to our understanding of the role of microglia in the pathogenesis of AUDs.
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影响因子：
9.3
作者：
Cherry JD;Olschowka JA;O'Banion MK
通讯作者：
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影响因子：
10.6
作者：
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影响因子：
3
作者：
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