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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >Effects of homocysteine on metabolic pathways in cultured astrocytes.
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Effects of homocysteine on metabolic pathways in cultured astrocytes.

机译：同型半胱氨酸对培养的星形胶质细胞代谢途径的影响。

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Homocysteine is an amino acid that is an important risk factor for several neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Increased homocysteine levels induce neuronal cell death in a variety of neuronal types. However, very few studies have probed the effects of homocysteine in astrocytes. The present study investigated the effects of homocysteine on primary cultures of astrocytes by exposing astrocytes to 400muM homocysteine for 20h. Metabolic extracts of cells were prepared following a 4-h incubation in minimum medium with 5.5mM [U-(13)C]glucose in the presence or absence of homocysteine and analysed using (13)C NMR. The expression level of pyruvate dehydrogenase kinase isoform 2 (PDK-2), NAD(P)H levels and mitochondrial membrane potential responses were investigated following culture with homocysteine. Metabolomic analysis was performed using (1)H NMR spectroscopy and pattern recognition analysis. Following incubation with homocysteine there was a significant decrease (48%)in the ratio of flux through pyruvate carboxylase (PC) and pyruvate dehydrogenase (PDH) which was due to an increased flux through PDH. In addition, homocysteine culture resulted in a significant reduction in PDK-2 protein expression. Following stimulation with glucose there was a significant increase in NAD(P)H levels and an impaired hyperpolarisation of the mitochondrial membrane in homocysteine-treated cells. Metabolomic analysis showed that the most discriminating metabolites following homocysteine treatment were choline and hypotaurine. In summary, the results demonstrated that sub-lethal concentrations of homocysteine caused significant metabolic changes and altered mitochondrial function in primary cultures of astrocytes.
机译：同型半胱氨酸是氨基酸，它是多种神经退行性疾病（例如阿尔茨海默氏病和帕金森氏病）的重要危险因素。同型半胱氨酸水平升高会导致多种神经元类型的神经元细胞死亡。但是，很少有研究探讨同型半胱氨酸在星形胶质细胞中的作用。本研究通过将星形胶质细胞暴露于400μM同型半胱氨酸20h来研究同型半胱氨酸对星形胶质细胞原代培养的影响。在存在或不存在同型半胱氨酸的情况下，在含有5.5mM [U-（13）C]葡萄糖的基本培养基中孵育4小时后，制备细胞的代谢提取物，并使用（13）C NMR进行分析。用高半胱氨酸培养后，研究了丙酮酸脱氢酶激酶同工型2（PDK-2）的表达水平，NAD（P）H水平和线粒体膜电位反应。代谢组学分析使用（1）H NMR光谱和模式识别分析进行。与高半胱氨酸孵育后，丙酮酸羧化酶（PC）和丙酮酸脱氢酶（PDH）的通量比率显着下降（48％），这是由于通过PDH的通量增加所致。另外，高半胱氨酸培养导致PDK-2蛋白表达显着降低。葡萄糖刺激后，同型半胱氨酸处理的细胞中NAD（P）H水平显着增加，线粒体膜的超极化受损。代谢组学分析显示，同型半胱氨酸治疗后最具区分性的代谢产物是胆碱和次牛磺酸。总之，结果表明亚致死浓度的高半胱氨酸在星形胶质细胞的原代培养物中引起了显着的代谢变化并改变了线粒体功能。

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