Gofman Larisa, Fernandes Nicole C, Potula Raghava
Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, PA, USA Center for Substance Abuse Research, Temple University School of Medicine, Philadelphia, PA, USA.
Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, PA, USA.
Alcohol Alcohol. 2016 Nov;51(6):647-654. doi: 10.1093/alcalc/agw009. Epub 2016 Mar 5.
Previously we have demonstrated altered microglia P2X4R expression in response to alcohol and pharmacological blockade with a selective P2X4R antagonist can reverse the action, suggesting that P2X4R play a role in mediating alcohol-induced effects on microglia. In the present study, we investigated the underlying signaling mediators, which may play a role in modulating P2X4R expression in microglia cells in response to alcohol.
Embryonic stem cell-derived microglia (ESdM) cells were used to investigate the potential mechanisms involved in the regulation of P2X4R in response to alcohol. Selective P2X4R antagonist and kinase inhibitors were used to further corroborate the signal transduction pathway through which alcohol modulates P2X4R expression in microglia.
Alcohol (100 mM) suppressed phosphorylated AKT and ERK cascades in native ESdM cells. This alcohol-induced suppression was confirmed to be P2X4R-dependent through the use of a selective P2X4R antagonist and knockdown of P2XR4 by siRNA. Alcohol increased transcriptional activity of CREB. P2X4R antagonist blocked alcohol-induced effects on CREB, suggesting a P2X4R-mediated effect.
These findings provide important clues to the underlying mechanism of purinoceptors in alcohol-induced microglia immune suppression.
此前我们已经证明，小胶质细胞P2X4R的表达会因酒精而改变，使用选择性P2X4R拮抗剂进行药理学阻断可逆转这种作用，这表明P2X4R在介导酒精对小胶质细胞的影响中发挥作用。在本研究中，我们调查了潜在的信号传导介质，它们可能在调节小胶质细胞中P2X4R表达以响应酒精方面发挥作用。
使用胚胎干细胞衍生的小胶质细胞（ESdM）来研究参与调节P2X4R以响应酒精的潜在机制。使用选择性P2X4R拮抗剂和激酶抑制剂来进一步证实酒精调节小胶质细胞中P2X4R表达的信号转导途径。
酒精（100 mM）抑制了天然ESdM细胞中磷酸化的AKT和ERK级联反应。通过使用选择性P2X4R拮抗剂和siRNA敲低P2XR4，证实这种酒精诱导的抑制是P2X4R依赖性的。酒精增加了CREB的转录活性。P2X4R拮抗剂阻断了酒精对CREB的诱导作用，表明这是一种P2X4R介导的效应。
这些发现为嘌呤受体在酒精诱导的小胶质细胞免疫抑制中的潜在机制提供了重要线索。
