Do β-Cells Generate Peroxynitrite in Response to Cytokine Treatment?
Do β-Cells Generate Peroxynitrite in Response to Cytokine Treatment?
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DOI：
10.1074/jbc.m113.522243
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发表时间：
2013-12-20
影响因子：
4.8
通讯作者：
Corbett, John A.
中科院分区：
文献类型：
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作者：
Broniowska, Katarzyna A.;Mathews, Clayton E.;Corbett, John A.
背景：介导细胞因子诱导的细胞死亡的物质是一氧化氮还是过氧亚硝酸盐，目前尚不清楚。结果：细胞因子刺激一氧化氮的形成，但不能刺激-细胞产生过氧亚硝酸盐。结论：终末分化细胞与巨噬细胞不同，不产生过氧亚硝酸盐。意义：过氧亚硝酸盐不会导致细胞因子诱导的细胞损伤。本研究的目的是确定负责细胞因子介导的细胞死亡的反应性物种。诱导型一氧化氮合酶抑制剂可防止这种死亡，而使用供体添加外源性一氧化氮可诱导细胞死亡。一氧化氮与超氧化物的反应产生过氧亚硝酸盐，这种强大的氧化剂被认为是细胞因子治疗反应中细胞死亡的中介。近年来，香豆素-7-硼酸盐作为一种选择性检测过氧亚硝酸盐的探针得到了发展。使用该试剂，我们发现在产生一氧化氮的巨噬细胞中加入NADPH氧化酶激活剂phorbol 12-肉豆酸酯13-醋酸酯会导致过氧亚硝酸盐的产生。使用类似的方法，我们证明细胞因子不能刺激大鼠胰岛细胞和胰岛素瘤细胞产生过氧亚硝酸盐，无论是否使用phorbol 12-肉豆蔻酸13-乙酸处理。当使用氧化还原循环器被迫产生超氧化物时，这一生成与保护一氧化氮毒性有关。这些发现表明：(i)一氧化氮可能是细胞因子毒性作用的介质，（ii）细胞不产生过氧亚硝酸盐以响应细胞因子，（iii）当被迫产生超氧化物时，超氧化物清除一氧化氮与保护细胞免受一氧化氮介导的毒性有关。
Background: The species that mediates cytokine-induced -cell death, be it nitric oxide or peroxynitrite, is unknown. Results: Cytokines stimulate the formation of nitric oxide but fail to stimulate peroxynitrite production by -cells. Conclusion: Terminally differentiated -cells, unlike macrophages, do not produce peroxynitrite. Significance: Peroxynitrite does not contribute to cytokine-induced -cell damage.The purpose of this study was to determine the reactive species that is responsible for cytokine-mediated -cell death. Inhibitors of inducible nitric oxide synthase prevent this death, and addition of exogenous nitric oxide using donors induces -cell death. The reaction of nitric oxide with superoxide results in the generation of peroxynitrite, and this powerful oxidant has been suggested to be the mediator of -cell death in response to cytokine treatment. Recently, coumarin-7-boronate has been developed as a probe for the selective detection of peroxynitrite. Using this reagent, we show that addition of the NADPH oxidase activator phorbol 12-myristate 13-acetate to nitric oxide-producing macrophages results in peroxynitrite generation. Using a similar approach, we demonstrate that cytokines fail to stimulate peroxynitrite generation by rat islets and insulinoma cells, either with or without phorbol 12-myristate 13-acetate treatment. When forced to produce superoxide using redox cyclers, this generation is associated with protection from nitric oxide toxicity. These findings indicate that: (i) nitric oxide is the likely mediator of the toxic effects of cytokines, (ii) -cells do not produce peroxynitrite in response to cytokines, and (iii) when forced to produce superoxide, the scavenging of nitric oxide by superoxide is associated with protection of -cells from nitric oxide-mediated toxicity.