Copetti-Santos Daniela, Moraes Vitoria, Weiler Dácio Franco, de Mello Alexandre Silva, Machado Fernanda de Souza, Marinho Jéssica Pereira, Siebert Cassiana, Kolling Janaina, Funchal Cláudia, Wyse Angela T S, Coelho Janice Carneiro
Biochemistry Department, Lysosomal Storage Diseases Laboratory, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.
Biosciences and Rehabilitation, Centro Universitário Metodista (IPA), Porto Alegre, RS, Brazil.
Lipids. 2015 Oct;50(10):937-44. doi: 10.1007/s11745-015-4062-4. Epub 2015 Sep 7.
The objective of this study was to determine the effect of U18666A, an inhibitor of cholesterol synthesis and its intracellular transport, on oxidative stress parameters in cortical astrocytes cultured from Wistar rats (0-3 days old). The cultures were incubated with U18666A (0.25 µg/mL) for 48 h, conditions that are considered ideal to mimic Niemann-Pick type C disease. A variety of indicators of oxidative stress were measured. U18666A treatment increased cholesterol 2-fold in treated compared to control astrocytes. Oxidative stress was significantly elevated in treated cells as demonstrated by a 1.7-fold increase in thiobarbituric acid reactive substances, a 60% decrease is sulfhydral groups, and a 3.7-fold increase in carbonyl groups, indicative of increased lipid and protein oxidation following U18666A treatment. Consistent with these changes, both catalase and superoxide dismutase activities were significantly reduced nearly 50% in treated compared to control astrocytes. Collectively, these change resulted in a 50% reduction in Na(+), K(+)-ATPase specific activity following U18666A treatment, suggesting a significant alteration in its plasma membrane environment. Overall, these changes indicate that U18666A treatment results in increased cholesterol levels and an increased level of oxidative stress in cortical astrocytes, consistent with what is observed in Niemann-Pick type C disease.
本研究的目的是确定胆固醇合成及其细胞内转运抑制剂U18666A对从Wistar大鼠（0 - 3日龄）培养的皮质星形胶质细胞氧化应激参数的影响。将培养物与U18666A（0.25 µg/mL）孵育48小时，这些条件被认为是模拟尼曼-匹克C型病的理想条件。测量了多种氧化应激指标。与对照星形胶质细胞相比，U18666A处理使处理后的细胞胆固醇增加了2倍。处理后的细胞氧化应激显著升高，硫代巴比妥酸反应性物质增加了1.7倍、巯基减少了60%、羰基增加了3.7倍，表明U18666A处理后脂质和蛋白质氧化增加。与这些变化一致，与对照星形胶质细胞相比，处理后的过氧化氢酶和超氧化物歧化酶活性均显著降低近50%。总体而言，这些变化导致U18666A处理后Na(+)，K(+)-ATP酶比活性降低了50%，表明其质膜环境发生了显著改变。总的来说，这些变化表明U18666A处理导致皮质星形胶质细胞胆固醇水平升高和氧化应激水平增加，这与尼曼-匹克C型病中观察到的情况一致。
