Neuroglobin overexpression inhibits oxygen-glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons.
Neuroglobin overexpression inhibits oxygen-glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons.
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DOI：
10.1016/j.nbd.2013.04.015
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发表时间：
2013-08
影响因子：
6.1
通讯作者：
Wang X
中科院分区：
文献类型：
--
作者：
Yu Z;Liu N;Li Y;Xu J;Wang X
脑红蛋白(NGB)是一种内源性神经保护分子，对缺氧/缺血性脑损伤具有保护作用，但其作用机制尚不清楚。我们最近的研究发现，NGB可以与线粒体通透性转换(MPT)调节因子--电压依赖性阴离子通道(VDAC)结合。在这项研究中，我们研究了NGB在原代培养的小鼠皮质神经元缺氧缺糖(OGD)后MPT孔(MPTP)开放中的作用。免疫共沉淀(Co-IP)和免疫细胞化学结果显示，缺氧后NGB与VDAC的结合增强，说明OGD增强了NGB-VDAC的相互作用。NGB过表达保护原代培养的小鼠皮质神经元免受OGD诱导的神经元死亡，其程度与MPTP开放抑制剂环孢素A(CsA)预处理相当。我们进一步通过NGB过表达和敲除方法在原代培养的神经元中检测了NGB在OGD诱导的MPTP开放中的作用，并将重组的NGB暴露于分离的线粒体。与CsA处理相同，NGB过表达显著降低OGD诱导的原代培养神经元线粒体肿胀、线粒体NAD+释放和细胞色素c(Cytc)释放等MPTP开放标志物。重组NGB孵育可显著降低OGD诱导的线粒体NAD+释放和Cytc释放。相反，NGB基因敲除显著增加了OGD诱导的神经元死亡，也增加了OGD诱导的线粒体NAD+释放和Cytc释放，这些结果可被CsA预处理所挽救。综上所述，我们的结果表明，NGB过表达可以抑制OGD诱导的原代培养小鼠皮质神经元MPTP的开放，这可能是NGB神经保护的分子机制之一。
Neuroglobin (Ngb) is an endogenous neuroprotective molecule against hypoxic/ischemic brain injury, but the underlying mechanisms remain largely undefined. Our recent study revealed that Ngb can bind to voltage-dependent anion channel (VDAC), a regulator of mitochondria permeability transition (MPT). In this study we examined the role of Ngb in MPT pore (mPTP) opening following oxygen-glucose deprivation (OGD) in primary cultured mouse cortical neurons. Co-immunoprecipitation (Co-IP) and immuocytochemistry showed that the binding between Ngb and VDAC was increased after OGD compared to normoxia, indicating the OGD-enhanced Ngb-VDAC interaction. Ngb overexpression protected primary mouse cortical neurons from OGD-induced neuronal death, to an extent comparable to mPTP opening inhibitor, cyclosporine A (CsA) pretreatment. We further measured the role of Ngb in OGD-induced mPTP opening using Ngb overexpression and knockdown approaches in primary cultured neurons, and recombinant Ngb exposure to isolated mitochondria. Same as CsA pretreatment, Ngb overexpression significantly reduced OGD-induced mPTP opening markers including mitochondria swelling, mitochondrial NAD+ release, and cytochrome c (Cyt c) release in primary cultured neurons. Recombinant Ngb incubation significantly reduced OGD-induced NAD+ release and Cyt c release from isolated mitochondria. In contrast, Ngb knockdown significantly increased OGD-induced neuron death, and increased OGD-induced mitochondrial NAD+ release and Cyt c release as well, and these outcomes could be rescued by CsA pretreatment. In summary, our results demonstrated that Ngb overexpression can inhibit OGD-induced mPTP opening in primary cultured mouse cortical neurons, which may be one of the molecular mechanisms of Ngb's neuroprotection.
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10.1152/ajpheart.01012.2008
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影响因子：
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