The noble gas argon modifies extracellular signal-regulated kinase 1/2 signaling in neurons and glial cells
The noble gas argon modifies extracellular signal-regulated kinase 1/2 signaling in neurons and glial cells
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DOI：
10.1016/j.ejphar.2011.10.045
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发表时间：
2012-01-15
影响因子：
5
通讯作者：
Coburn, Mark
中科院分区：
文献类型：
--
作者：
Fahlenkamp, Astrid V.;Rossaint, Rolf;Coburn, Mark
最近，惰性气体氩已被确定为一种有效的神经保护剂，但对其细胞效应知之甚少。在这项体外研究中，我们研究了氩气对细胞外信号调节激酶（ERK）1/2的影响，细胞外信号调节激酶（ERK）1/2是一种普遍存在的酶，在细胞增殖和生存中具有多种功能。将原代神经元和星形胶质细胞培养物以及小胶质细胞系BV-2暴露于50 vol. %氩气在用50 ng/ml LPS刺激小胶质细胞后研究了进一步的可能作用。通过磷酸化状态特异性蛋白质印迹法评估ERK 1/2活化，通过实时PCR和蛋白质印迹法评估细胞因子水平。总磷酸酪氨酸磷酸酶活性与对硝基苯基磷酸盐进行了检查。氩暴露30 min后，小胶质细胞ERK 1/2信号显著激活。在氩暴露后星形胶质细胞和神经元中也发现ERK 1/2磷酸化增强，但缺乏统计学意义。在小胶质细胞中，氩气基本上不干扰LPS诱导的ERK 1/2激活和炎性细胞因子诱导。加入MEK抑制剂U 0126可消除诱导的ERK 1/2磷酸化。细胞磷酸酶活性和磷酸化ERK 1/2的失活没有被氩气改变。总之，氩气增强ERK 1/2活性的小胶质细胞通过上游激酶MEK，可能是通过直接激活模式。在体外星形胶质细胞和神经元中的ERK 1/2信号也受到影响，尽管没有统计学意义。氩气激活ERK 1/2是否会影响体内脑中的分化和存活等细胞功能，必须在未来的实验中确定。(C)2011爱思唯尔有限公司版权所有。
Recently, the noble gas argon has been identified as a potent neuroprotective agent, but little is known about its cellular effects. In this in vitro study, we investigated argon's influence on the extracellular signal-regulated kinase (ERK) 1/2, a ubiquitous enzyme with numerous functions in cell proliferation and survival. Primary neuronal and astroglial cell cultures and the microglial cell line BV-2 were exposed to 50 vol.% argon. Further possible effects were studied following stimulation of microglia with 50 ng/ml LPS. ERK 1/2 activation was assessed by phosphorylation state-specific western blotting, cytokine levels by real-time PCR and western blotting. Total phosphotyrosine phosphatase activity was examined with p-nitrophenylphosphate. After 30 min exposure, argon significantly activated ERK 1/2 signaling in microglia. Enhanced phosphorylation of ERK 1/2 was also found in astrocytes and neurons following argon exposure, but it lacked statistical significance. In microglia, argon did not substantially interfere with LPS-induced ERK1/2 activation and inflammatory cytokine induction. Addition of the MEK-Inhibitor U0126 abolished the induced ERK 1/2 phosphorylation. Cellular phosphatase activity and the inactivation of phosphorylated ERK 1/2 were not altered by argon. In conclusion, argon enhanced ERK 1/2 activity in microglia via the upstream kinase MEK, probably through a direct mode of activation. ERK 1/2 signaling in astrocytes and neurons in vitro was also influenced, although not with statistical significance. Whether ERK 1/2 activation by argon affects cellular functions like differentiation and survival in the brain in vivo will have to be determined in future experiments. (C) 2011 Elsevier B.V. All rights reserved.