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Mechanism for generation of hydrogen peroxide and change of mitochondrial membrane potential during rotenone-induced apoptosis.

机译:鱼藤酮诱导的细胞凋亡过程中过氧化氢的产生和线粒体膜电位的变化机制。

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Rotenone, an inhibitor of NADH dehydrogenase complex, is a naturally occurring insecticide, which is capable of inducing apoptosis. Rotenone-induced apoptosis is considered to contribute to its anticancer effect and the etiology of Parkinson's disease (PD). We demonstrated that rotenone induced internucleosomal DNA fragmentation, DNA ladder formation, in human cultured cells, HL-60 (promyelocytic leukemia) and BJAB cells (B-cell lymphoma). Flow cytometry showed that rotenone induced H(2)O(2) generation, followed by significant changes in the mitochondrial membrane potential (DeltaPsim). Caspase-3 activity increased in HL-60 cells in a time-dependent manner. These apoptotic events were delayed in HP100 cells, an H(2)O(2)-resistant clone of HL-60, confirming the involvement of H(2)O(2) in apoptosis. Expression of anti-apoptotic protein, Bcl-2, in BJAB cells drastically inhibited DeltaPsim change and DNA ladder formation but not H(2)O(2) generation, confirming the participation of mitochondrial dysfunction in apoptosis. NAD(P)H oxidase inhibitors prevented H(2)O(2) generation and DNA ladder formation. These results suggest that rotenone induces O(2)(-)-derived H(2)O(2) generation through inhibition of NADH dehydrogenase complex and/or activation of NAD(P)H oxidase, and H(2)O(2) generation causes the disruption of mitochondrial membrane in rotenone-induced apoptosis.
机译:鱼藤酮是NADH脱氢酶复合物的抑制剂,是一种天然杀虫剂,能够诱导细胞凋亡。鱼藤酮诱导的细胞凋亡被认为有助于其抗癌作用和帕金森氏病(PD)的病因。我们证明鱼藤酮在人类培养的细胞,HL-60(早幼粒细胞白血病)和BJAB细胞(B细胞淋巴瘤)中诱导核小体间DNA片段化,DNA阶梯形成。流式细胞仪显示鱼藤酮诱导H(2)O(2)代,然后在线粒体膜电位(DeltaPsim)的显着变化。 Caspase-3活性在HL-60细胞中呈时间依赖性增加。这些凋亡事件在HP100细胞,HL-60的H(2)O(2)耐药克隆中被延迟,证实H(2)O(2)参与凋亡。 BJAB细胞中抗凋亡蛋白Bcl-2的表达大大抑制了DeltaPsim的变化和DNA梯形的形成,但没有抑制H(2)O(2)的生成,从而证实了线粒体功能障碍参与了细胞凋亡。 NAD(P)H氧化酶抑制剂阻止了H(2)O(2)的产生和DNA阶梯的形成。这些结果表明鱼藤酮通过抑制NADH脱氢酶复合物和/或激活NAD(P)H氧化酶和H(2)O(2)诱导O(2)(-)衍生的H(2)O(2)生成。 )的生成导致鱼藤酮诱导的细胞凋亡中线粒体膜的破坏。

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