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首页> 美国卫生研究院文献>Biochemical Journal >Peroxynitrite modulates tyrosine phosphorylation and phosphoinositide signalling in human neuroblastoma SH-SY5Y cells: attenuated effects in human 1321N1 astrocytoma cells.
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Peroxynitrite modulates tyrosine phosphorylation and phosphoinositide signalling in human neuroblastoma SH-SY5Y cells: attenuated effects in human 1321N1 astrocytoma cells.

机译：过氧亚硝酸盐调节人神经母细胞瘤SH-SY5Y细胞中的酪氨酸磷酸化和磷酸肌醇信号转导：在人类1321N1星形细胞瘤细胞中的减弱作用。

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Peroxynitrite may contribute to oxidative stress involving neurodegeneration in several disorders, including Alzheimer's disease. As with other reactive oxygen species, peroxynitrite might affect neuronal signalling systems, actions that could contribute to adaptive or deleterious cellular outcomes, but such effects have not previously been studied. To address this issue directly, peroxynitrite (50-500 microM) was administered to human neuroblastoma SH-SY5Y cells to assess its effects on protein tyrosine nitration, phosphoinositide signalling and protein tyrosine phosphorylation. Peroxynitrite rapidly increased the nitrotyrosine immunoreactivity of numerous proteins, primarily in the cytosol. Peroxynitrite inhibited, in a concentration-dependent manner, phosphoinositide hydrolysis stimulated by activation of muscarinic receptors with carbachol and the inhibition was greater after the depletion of cellular glutathione. In comparison, muscarinic receptor-stimulated phosphoinositide hydrolysis in human astrocytoma 1321N1 cells was less vulnerable to inhibition by peroxynitrite either without or with prior depletion of glutathione. There was a large, rapid and reversible increase in the tyrosine phosphorylation of the p120 Src substrate in peroxynitrite-treated SH-SY5Y cells, a response that was potentiated by glutathione depletion; in contrast, peroxynitrite decreased the tyrosine phosphorylation of focal adhesion kinase and paxillin. Tyrosine phosphorylation of p120 in 1321N1 astrocytoma cells was less sensitive to modulation by peroxynitrite. Thus alterations in phosphoinositide signalling and protein tyrosine phosphorylation were greater in neuroblastoma than astrocytoma cells, and modulation of these signalling processes probably contributes to neuronal mechanisms of the response to peroxynitrite.
机译：过氧亚硝酸盐可能导致包括阿尔茨海默氏病在内的多种疾病中涉及神经变性的氧化应激。与其他活性氧一样，过亚硝酸盐可能会影响神经元信号传导系统，这些行为可能有助于适应性或有害性细胞结果，但以前尚未研究过这种作用。为了直接解决这个问题，将过氧亚硝酸盐（50-500 microM）应用于人神经母细胞瘤SH-SY5Y细胞，以评估其对蛋白质酪氨酸硝化，磷酸肌醇信号传导和蛋白质酪氨酸磷酸化的影响。过氧亚硝酸盐迅速增加了许多蛋白质（主要是在细胞质中）的硝基酪氨酸免疫反应性。过氧亚硝酸盐以浓度依赖的方式抑制由毒蕈碱受体用碳酰胆碱活化而刺激的磷酸肌醇水解，并且在细胞内谷胱甘肽耗竭后，抑制作用更大。相比之下，人星形细胞瘤1321N1细胞中毒蕈碱受体刺激的磷酸肌醇水解在不使用或事先耗尽谷胱甘肽的情况下较不易受到过氧亚硝酸盐的抑制。在过氧亚硝酸盐处理的SH-SY5Y细胞中，p120 Src底物的酪氨酸磷酸化有大量，快速和可逆的增加，谷胱甘肽耗竭增强了这种反应。相反，过氧亚硝酸盐减少了粘着斑激酶和Paxillin的酪氨酸磷酸化。在1321N1星形细胞瘤细胞中p120的酪氨酸磷酸化对过亚硝酸盐的调节较不敏感。因此，在神经母细胞瘤中，磷酸肌醇信号转导和蛋白酪氨酸磷酸化的改变要比星形细胞瘤细胞大，并且这些信号转导过程的调节可能有助于对过亚硝酸盐反应的神经元机制。

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