D-β-hydroxybutyrate protects dopaminergic SH-SY5Y cells in a rotenone model of Parkinson's disease
D-β-hydroxybutyrate protects dopaminergic SH-SY5Y cells in a rotenone model of Parkinson's disease
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DOI：
10.1002/jnr.21021
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发表时间：
2006-11-01
影响因子：
4.2
通讯作者：
Nakashima, Kenji
中科院分区：
文献类型：
--
作者：
Imamura, Keiko;Takeshima, Takao;Nakashima, Kenji
据推测，帕金森病（PD）的发病机制与线粒体功能障碍有关。鱼藤酮是线粒体复合物I的抑制剂，提供了体内和体外PD模型。我们使用SH-SY 5 Y多巴胺能神经母细胞瘤细胞研究了D-β-羟基丁酸（bHB）（一种酮体）对鱼藤酮毒性的神经保护作用。将通过全反式视黄酸分化的SH-SY 5 Y细胞暴露于浓度范围为0至1，000 nM的鱼藤酮。我们通过alamarBlue测定法评价细胞氧化还原，通过乳酸脱氢酶（LDH）测定法评价细胞活力，通过活/死测定法评价存活/死亡比。暴露于鱼藤酮48小时氧化细胞，并以浓度依赖性方式降低其活力和存活率。用8 mM bHB预处理细胞对SH-SY 5 Y细胞提供了显著的保护。而鱼藤酮引起的线粒体膜电位的损失，释放细胞色素c到胞质溶胶中，并减少细胞色素c的含量在线粒体中，除了bHB阻断这种毒性作用。bHB还减弱鱼藤酮诱导的caspase-9和caspase-3的活化。施用0-10 mM 3-硝基丙酸（复合物II抑制剂）也降低了通过alamarBlue测定法测量的SH-SY 5 Y细胞的还原能力。用8 mM bHB预处理减弱了alamarBlue荧光的降低。这些数据表明，bHB具有神经保护作用，通过逆转复合物I或II的抑制来支持线粒体呼吸系统。酮体是哺乳动物大脑中的替代能量来源，似乎在PD中具有治疗潜力。(c)2006威利-利斯公司
It has been postulated that the pathogenesis of Parkinson's disease (PD) is associated with mitochondrial dysfunction. Rotenone, an inhibitor of mitochondrial complex I, provides models of PD both in vivo and in vitro. We investigated the neuroprotective effect of D-beta-hydroxybutyrate (bHB), a ketone body, against rotenone toxicity by using SH-SY5Y dopaminergic neuroblastoma cells. SH-SY5Y cells, differentiated by all-trans-retinoic acid, were exposed to rotenone at concentrations ranging from 0 to 1,000 nM. We evaluated cellular oxidation reduction by the alamarBlue assay, viability by lactate dehydrogenase (LDH) assay, and survival/death ratio by live/dead assays. Exposure to rotenone for 48 hr oxidized cells and decreased their viability and survival rate in a concentration-dependent manner. Pretreatment of cells with 8 mM bHB provided significant protection to SH-SY5Y cells. Whereas rotenone caused the loss of mitochondrial membrane potential, released cytochrome c into the cytosol, and reduced cytochrome c content in mitochondria, addition of bHB blocked this toxic effect. bHB also attenuated the rotenone-induced activation of caspase-9 and caspase-3. Administration of 0-10 mM 3-nitropropionic acid, a complex II inhibitor, also decreased the reducing power of SH-SY5Y cells measured by alamarBlue assay. Pretreatment with 8 mM bHB attenuated the decrease of alamarBlue fluorescence. These data demonstrated that bHB had a neuroprotective effect that supported the mitochondrial respiration system by reversing the inhibition of complex I or II. Ketone bodies, the alternative energy source in the mammalian brain, appear to have therapeutic potential in PD. (c) 2006 Wiley-Liss, Inc.