Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastoma cells:: role of ERK1/2 in H2O2-induced cell death
Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastoma cells:: role of ERK1/2 in H2O2-induced cell death
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DOI：
10.1016/j.ejphar.2003.10.032
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发表时间：
2004-01-12
影响因子：
5
通讯作者：
Dickenson, JM
中科院分区：
文献类型：
--
作者：
Ruffels, J;Griffin, M;Dickenson, JM
包括H2 O2在内的活性氧物质激活一系列与细胞死亡和细胞生存途径密切相关的细胞内信号级联反应。人神经母细胞瘤SH-SY 5 Y细胞系被广泛用作研究氧化应激诱导的神经元细胞死亡的模型细胞系统。然而，目前很少有人知道过氧化氢在SH-SY 5 Y细胞中激活的信号通路。因此，在本研究中，我们研究了过氧化氢对细胞外信号调节激酶1/2（ERK 1/2），c-Jun N-末端激酶（JNK），p38丝裂原活化蛋白激酶（p38 MAPK）和蛋白激酶B（PKB）在未分化和分化的SH-SY 5 Y细胞中的活化的影响。H2 O2刺激SH-SY 5 Y细胞ERK 1/2、JNK和PKB磷酸化的时间和浓度增加。H2 O2处理后p38 MAPK磷酸化没有增加。磷脂酰肌醇3-激酶（PI-3 K）抑制剂wortmannin和LY 294002（（2-（4-morpholinyl）-8-phenyl-4 H-1-benzopyran-4-one））抑制H2 O2诱导的ERK 1/2和PKB磷酸化的增加。此外，H2 O2介导的ERK 1/2活化的增加对MAPK激酶1（MEK 1）抑制剂PD 98059（2 '-氨基-3'-甲氧基黄酮）敏感，而JNK反应被JNK抑制剂SP 600125（蒽[1-9-cd]吡唑-6（2 H）-酮）阻断。用H2 O2（1 mM; 16 h）处理SH-SY 5 Y细胞显著增加了乳酸脱氢酶（LDH）向培养基中的释放，表明细胞活力降低。用渥曼青霉素、SP 600125或SB 203580（4-（4-氟苯基）-2-（4-甲基亚磺酰基苯基）-5-（4-吡啶基）1H-咪唑; p38 MAPK抑制剂）预处理对H2 O2诱导的未分化或分化SH-SY 5 Y细胞LDH释放无影响。相比之下，PD 98059和LY 294002显著降低未分化和分化的SH-SY 5 Y细胞中H2 O2诱导的细胞死亡。总之，我们已经表明，H2 O2刺激了未分化和分化的SH-SY 5 Y细胞中ERK 1/2，JNK和PKB的稳健增加。此外，所呈现的数据清楚地表明，ERK 1/2通路的抑制保护SH-SY 5 Y细胞免于H2 O2诱导的细胞死亡。(C)2003 Elsevier B. V.保留所有权利。
Reactive oxygen species including H2O2 activate an array of intracellular signalling cascades that are closely associated with cell death and cell survival pathways. The human neuroblastoma SH-SY5Y cell line is widely used as model cell system for studying neuronal cell death induced by oxidative stress. However, at present very little is known about the signalling pathways activated by H2O2 in SH-SY5Y cells. Therefore, in this study we have investigated the effect of H2O2 on extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38 MAPK) and protein kinase B (PKB) activation in undifferentiated and differentiated SH-SY5Y cells. H2O2 stimulated time and concentration increases in ERK1/2, JNK and PKB phosphorylation in undifferentiated and differentiated SH-SY5Y cells. No increases in p38 MAPK phosphorylation were observed following H2O2 treatment. The phosphatidylinositol 3-kinase (PI-3K) inhibitors wortmannin and LY 294002 ((2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one) inhibited H2O2-induced increases in ERK1/2 and PKB phosphorylation. Furthermore, H2O2-mediated increases in ERK1/2 activation were sensitive to the MAPK kinase 1 (MEK1) inhibitor PD 98059 (2'-amino-3'-methoxyflavone), whereas JNK responses were blocked by the JNK inhibitor SP 600125 (anthra[1-9-cd]pyrazol-6(2H)-one). Treatment of SH-SY5Y cells with H2O2 (1 mM; 16 h) significantly increased the release of lactate dehydrogenase (LDH) into the culture medium indicative of a decrease in cell viability. Pre-treatment with wortmannin, SP 600125 or SB 203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole; p38 MAPK inhibitor) had no effect on H2O2-induced LDH release from undifferentiated or differentiated SH-SY5Y cells. In contrast, PD 98059 and LY 294002 significantly decreased H2O2-induced cell death in both undifferentiated and differentiated SH-SY5Y cells. In conclusion, we have shown that H2O2 stimulates robust increases in ERK1/2, JNK and PKB in undifferentiated and differentiated SH-SY5Y cells. Furthermore, the data presented clearly suggest that inhibition of the ERK1/2 pathway protects SH-SY5Y cells from H2O2-induced cell death. (C) 2003 Elsevier B.V. All rights reserved.