Prence E M, Chaturvedi P, Newburg D S
Division of Medical Genetics, Shriver Center for Mental Retardation, Waltham, Massachusetts, USA.
J Neurosci Res. 1996 Feb 1;43(3):365-71. doi: 10.1002/(SICI)1097-4547(19960201)43:3<365::AID-JNR11>3.0.CO;2-4.
Gaucher disease is the most common lysosomal glycosphingolipid storage disease; decreased activity of glucosylceramide beta-glucosidase (GCase) results in the accumulation of glucocerebroside (GlcCer) in macrophage-derived cells. The most devastating types of Gaucher disease also involve neuronopathology, thought to be mediated by intracellular GlcCer accumulation in the brain. In this study, we developed an in vitro neuronal cell model for accumulation of endogenous GlcCer to enable studies on the cellular basis for the neuronopathology of this disease. A human neuroblastoma cell line (SH-SY5Y) was selected because it produced appreciable GCase. When these cells were treated with conduritol B epoxide (CBE), a competitive, irreversible inhibitor of this enzyme, GCase levels fell precipitously, while other lysosomal hydrolase levels were unaffected. Relative to untreated control cells, the CBE-treated cells accumulated higher levels of GlcCer, but not other related glycolipids, over time. Thus, this in vitro system displayed many essential biological parameters relevant for studies on cellular events responsible for the neurologic damage that occurs in some types of Gaucher disease. This model should also be useful in investigations of the normal role of sphingolipids in neuronal cell function.
戈谢病是最常见的溶酶体糖鞘脂贮积病；葡糖脑苷脂β-葡萄糖苷酶（GCase）活性降低导致葡糖脑苷脂（GlcCer）在巨噬细胞衍生细胞中蓄积。最严重类型的戈谢病还涉及神经病理学，被认为是由大脑中细胞内GlcCer蓄积介导的。在本研究中，我们建立了一个用于内源性GlcCer蓄积的体外神经元细胞模型，以便在细胞基础上研究该疾病的神经病理学。选择了一种人神经母细胞瘤细胞系（SH-SY5Y），因为它能产生可观的GCase。当用该酶的竞争性不可逆抑制剂环氧康杜立醇（CBE）处理这些细胞时，GCase水平急剧下降，而其他溶酶体水解酶水平未受影响。随着时间的推移，相对于未处理的对照细胞，经CBE处理的细胞蓄积了更高水平的GlcCer，但没有蓄积其他相关糖脂。因此，这个体外系统显示出许多与研究某些类型戈谢病中发生的神经损伤相关的细胞事件相关的重要生物学参数。该模型在研究鞘脂在神经元细胞功能中的正常作用方面也应该是有用的。
