Chronic gestational exposure to ethanol impairs insulin-stimulated survival and mitochondrial function in cerebellar neurons
Chronic gestational exposure to ethanol impairs insulin-stimulated survival and mitochondrial function in cerebellar neurons
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DOI：
10.1007/s00018-002-8475-x
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发表时间：
2002-05-01
影响因子：
8
通讯作者：
Wands, JR
中科院分区：
文献类型：
--
作者：
de la Monte, SM;Wands, JR
孕期长期接触乙醇对大脑发育有深远的不利影响。在这方面，使用酒精暴露的体外模型的研究表明，胰岛素信号转导机制受损，与神经细胞凋亡增加和线粒体功能降低有关。为了确定这些发现与胎儿酒精综合征的相关性，我们使用慢性妊娠乙醇暴露的大鼠模型，研究了胰岛素刺激的神经元存活和线粒体功能的机制。在酒精暴露的幼鼠中，小脑半球发育不良，细胞凋亡增加。分离培养小脑神经元，选择性地评价胰岛素的反应性。孕期乙醇暴露抑制胰岛素刺激的神经元活性、线粒体功能、钙调素AM保留(膜完整性)和GAPDH的表达，并增加二氢核糖胺荧光(氧化应激)和促凋亡基因(P53、Fas受体和Fas配体)的表达。此外，乙醇暴露的幼鼠产生的神经元培养降低了胰岛素刺激的Akt、GSK-3β和BAD磷酸化水平，增加了非磷酸化(激活)的GSK-3β和BAD蛋白表达水平。综上所述，这些结果表明，长期酒精暴露会显著损害胰岛素刺激的中枢神经系统神经元的存活机制，胰岛素信号机制的异常持续到出生后早期，这对大脑发育至关重要。
Chronic gestational exposure to ethanol has profound adverse effects on brain development. In this regard, studies using in vitro models of ethanol exposure demonstrated impaired insulin signaling mechanisms associated with increased apoptosis and reduced mitochondrial function in neuronal cells. To determine the relevance of these findings to fetal alcohol syndrome, we examined mechanisms of insulin-stimulated neuronal survival and mitochondrial function using a rat model of chronic gestational exposure to ethanol. In ethanol-exposed pups, the cerebellar hemispheres were hypoplastic and exhibited increased apoptosis. Isolated cerebellar neurons were cultured to selectively evaluate insulin responsiveness. Gestational exposure to ethanol inhibited insulin-stimulated neuronal viability, mitochondrial function, Calcein AM retention (membrane integrity), and GAPDH expression, and increased dihydrorosamine fluorescence (oxidative stress) and pro-apoptosis gene expression (p53, Fas-receptor, and Fas-ligand). In addition, neuronal cultures generated from ethanol-exposed pups had reduced levels of insulin-stimulated Akt, GSK-3beta, and BAD phosphorylation, and increased levels of non-phosphorylated (activated) GSK-3beta and BAD protein expression. The aggregate results suggest that insulin-stimulated central nervous system neuronal survival mechanisms are significantly impaired by chronic gestational exposure to ethanol, and that the abnormalities in insulin signaling mechanisms persist in the early postnatal period, which is critical for brain development.