The role of microglial P2X7: modulation of cell death and cytokine release.
The role of microglial P2X7: modulation of cell death and cytokine release.
复制标题
DOI：
10.1186/s12974-017-0904-8
复制
发表时间：
2017-07-17
影响因子：
9.3
通讯作者：
Bhattacharya A
中科院分区：
文献类型：
--
作者：
He Y;Taylor N;Fourgeaud L;Bhattacharya A
ATP门控P2 X7是一种非选择性阳离子通道，参与广泛的细胞功能以及病理生理过程，包括神经病理性疼痛、免疫反应和神经炎症。尽管P2 X7在小胶质细胞中表达丰富，但其在神经炎症中的作用仍不清楚。从P0-2 C57 BL/6野生型或P2 X7敲除（P2 X7-/-）小鼠幼仔的皮质中分离出原代小胶质细胞。脂多糖、脂多糖加IFNγ或IL 4加IL 13用于使小胶质细胞处于促炎或抗炎状态。通过RNA测序和定量实时PCR测量静息或活化的小鼠和人小胶质细胞中的P2 rx 7表达水平。通过细胞计数试剂盒-8和免疫细胞化学测量小胶质细胞死亡，并通过Luminex多重测定或ELISA使用P2 X7激动剂BzATP或P2 X7拮抗剂A-804598检查野生型或P2 X7 −/−小胶质细胞中的小胶质细胞分泌。通过蛋白质印迹分析P2 X7信号传导。首先，我们证实了P2 rx 7在小鼠和人的原代小胶质细胞中组成型表达。此外，在促炎和抗炎条件下，小鼠小胶质细胞中P2 rx 7 mRNA水平下调。第二，P2 X7激动剂BzATP引起小鼠小胶质细胞的细胞死亡，而这种作用在基础和促炎条件下被P2 X7敲除或被A-804598抑制，这表明P2 X7在BzATP诱导的小胶质细胞死亡中的介导作用。第三，BzATP诱导的IL 1家族细胞因子（包括IL 1 α、IL 1 β和IL 18）的释放在P2 X7 −/−小胶质细胞中被阻断，或在促炎性小胶质细胞中被A-804598阻断，而其他细胞因子/趋化因子的释放与P2 X7激活无关。这些发现支持P2 X7在IL 1家族细胞因子释放中的特异性作用。最后，发现P2 X7激活与AKT和ERK通路相关，这可能是P2 X7在小胶质细胞中功能的潜在机制。这些结果表明P2 X7介导BzATP诱导的小胶质细胞死亡和IL 1家族细胞因子的特异性释放，表明P2 X7在神经炎症中的重要作用，并暗示靶向P2 X7用于治疗神经炎症性疾病的潜力。本文的在线版本（doi：10.1186/s12974-017-0904-8）包含补充材料，可供授权用户使用。
ATP-gated P2X7 is a non-selective cation channel, which participates in a wide range of cellular functions as well as pathophysiological processes including neuropathic pain, immune response, and neuroinflammation. Despite its abundant expression in microglia, the role of P2X7 in neuroinflammation still remains unclear. Primary microglia were isolated from cortices of P0-2 C57BL/6 wild-type or P2X7 knockout (P2X7−/−) mouse pups. Lipopolysaccharide, lipopolysaccharide plus IFNγ, or IL4 plus IL13 were used to polarize microglia to pro-inflammatory or anti-inflammatory states. P2rx7 expression level in resting or activated mouse and human microglia was measured by RNA-sequencing and quantitative real-time PCR. Microglial cell death was measured by cell counting kit-8 and immunocytochemistry, and microglial secretion in wild-type or P2X7−/− microglia was examined by Luminex multiplex assay or ELISA using P2X7 agonist BzATP or P2X7 antagonist A-804598. P2X7 signaling was analyzed by Western blot. First, we confirmed that P2rx7 is constitutively expressed in mouse and human primary microglia. Moreover, P2rx7 mRNA level was downregulated in mouse microglia under both pro- and anti-inflammatory conditions. Second, P2X7 agonist BzATP caused cell death of mouse microglia, while this effect was suppressed either by P2X7 knockout or by A-804598 under both basal and pro-inflammatory conditions, which suggests the mediating role of P2X7 in BzATP-induced microglial cell death. Third, BzATP-induced release of IL1 family cytokines including IL1α, IL1β, and IL18 was blocked in P2X7−/− microglia or by A-804598 in pro-inflammatory microglia, while the release of other cytokines/chemokines was independent of P2X7 activation. These findings support the specific role of P2X7 in IL1 family cytokine release. Finally, P2X7 activation was discovered to be linked to AKT and ERK pathways, which may be the underlying mechanism of P2X7 functions in microglia. These results reveal that P2X7 mediates BzATP-induced microglial cell death and specific release of IL1 family cytokines, indicating the important role of P2X7 in neuroinflammation and implying the potential of targeting P2X7 for the treatment of neuroinflammatory disorders. The online version of this article (doi:10.1186/s12974-017-0904-8) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
5.8
作者：
Mosher, Kira Irving;Wyss-Coray, Tony
通讯作者：
Wyss-Coray, Tony
巨噬细胞激活和极化改变 P2X7 受体分泌组，影响炎症过程。
影响因子：
4.6
作者：
de Torre-Minguela C;Barberà-Cremades M;Gómez AI;Martín-Sánchez F;Pelegrín P
通讯作者：
Pelegrín P
影响因子：
56.9
作者：
Nimmerjahn, A;Kirchhoff, F;Helmchen, F
通讯作者：
Helmchen, F
影响因子：
7.3
作者：
Bhattacharya, Anindya;Wang, Qi;Letavic, Michael A.
通讯作者：
Letavic, Michael A.
影响因子：
7.3
作者：
Able, S. L.;Fish, R. L.;Katugampola, S. D.
通讯作者：
Katugampola, S. D.