P2X4 receptor regulates alcohol-induced responses in microglia.
P2X4 receptor regulates alcohol-induced responses in microglia.
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DOI：
10.1007/s11481-014-9559-8
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发表时间：
2014-12
影响因子：
6.2
通讯作者：
Potula, Raghava
中科院分区：
文献类型：
--
作者：
Gofman, Larisa;Cenna, Jonathan M.;Potula, Raghava
越来越多的证据表明，酒精诱导的神经病理可能是由多细胞反应，其中小胶质细胞发挥了突出的作用。嘌呤能受体信号在调节小胶质细胞功能中起关键作用，更重要的是，介导酒精诱导的效应。我们的研究结果表明，酒精增加P2 X4受体（P2 X4 R）的表达，这改变了小胶质细胞的功能，包括钙动员，迁移和吞噬。我们的结果显示，与未处理的对照相比，在酒精处理48小时后，在胚胎干细胞衍生的小胶质细胞（ESdM）中，通过实时qPCR分析的P2 X4基因表达（* p <0.002）和通过流式细胞术分析的蛋白质表达（**p<0.004）显著上调。用P2 X4 R选择性拮抗剂5-BDBD发现乙醇处理的ESdM细胞中的钙动员是P2 X4 R依赖性的。与对照相比，在处理48小时后，酒精使小胶质细胞向fractalkine（CX 3CL 1）的迁移减少75%（*p<0.001）。使用拮抗剂5-BDBD证实CX 3CL 1依赖性迁移是P2 X4受体依赖性的，与单独的酒精相比，拮抗剂5-BDBD逆转了这种作用（*p<0.001）。类似地，与对照相比，48小时的酒精处理使小胶质细胞的吞噬作用显著降低15%（*p<0.05）。5-在酒精处理之前的BDBD预处理显著增加了小胶质细胞吞噬作用（*p<0.001）。与单独乙醇处理相比，用5-BDBD阻断P2 X4 R信号传导降低了钙动员水平。这些发现表明，P2 X4受体可能在酒精滥用的背景下调节小胶质细胞功能中发挥作用。
Mounting evidence indicates that alcohol-induced neuropathology may result from multicellular responses in which microglia cells play a prominent role. Purinergic receptor signaling plays a key role in regulating microglial function and, more importantly, mediates alcohol-induced effects. Our findings demonstrate that alcohol increases expression of P2X4 receptor (P2X4R), which alters the function of microglia, including calcium mobilization, migration and phagocytosis. Our results show a significant up-regulation of P2X4 gene expression as analyzed by real-time qPCR (***p<0.002) and protein expression as analyzed by flow cytometry (**p<0.004) in embryonic stem cell-derived microglial cells (ESdM) after 48 hours of alcohol treatment, as compared to untreated controls. Calcium mobilization in ethanol treated ESdM cells was found to be P2X4R dependent using 5-BDBD, a P2X4R selective antagonist. Alcohol decreased migration of microglia towards fractalkine (CX3CL1) by 75% following 48 hours of treatment compared to control (***p<0.001). CX3CL1-dependent migration was confirmed to be P2X4 receptor-dependent using the antagonist 5-BDBD, which reversed the effects as compared to alcohol alone (***p<0.001). Similarly, 48 hours of alcohol treatment significantly decreased phagocytosis of microglia by 15% compared to control (*p<0.05). 5-BDBD pre-treatment prior to alcohol treatment significantly increased microglial phagocytosis (***p<0.001). Blocking P2X4R signaling with 5-BDBD decreased the level of calcium mobilization compared to ethanol treatment alone. These findings demonstrate that P2X4 receptor may play a role in modulating microglial function in the context of alcohol abuse.
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DOI：
10.1186/1478-811x-11-12
发表时间：
2013-02-17
期刊：
Cell communication and signaling : CCS
影响因子：
--
作者：
Glaser T;Resende RR;Ulrich H
通讯作者：
Ulrich H
影响因子：
2.4
作者：
Dooley R;Mashukova A;Toetter B;Hatt H;Neuhaus EM
通讯作者：
Neuhaus EM
影响因子：
2.3
作者：
Aroor, AR;Baker, RC
通讯作者：
Baker, RC
影响因子：
6.1
作者：
Ko, WH;Au, CL;Yip, CY
通讯作者：
Yip, CY
影响因子：
3
作者：
Inoue, Kazuhide;Tsuda, Makoto
通讯作者：
Tsuda, Makoto