Vered A, Eugenio M, Arie D, Augusto P, Sergio M, Shimon G
Department of Membrane Biochemistry and Neurochemistry, Hebrew University, Hadassah School of Medicine, Jerusalem, Israel.
Clin Chim Acta. 1993 Sep 30;218(2):139-47. doi: 10.1016/0009-8981(93)90178-7.
Glycosphingolipids, labeled with the fluorescent probe lissamine rhodamine were administered to skin fibroblasts in culture and were hydrolyzed in the intact cells to the corresponding lissamine rhodamine ceramide (N-acylsphingosine). This fluorescent ceramide was converted in the intact cells to the corresponding sphingomyelin which was secreted into the culture medium. In comparison, ceramide is not formed in cells derived from patients with lipid storage diseases, because of deficiencies in lysosomal glycolipid hydrolases. Consequently, fluorescent sphingomyelin was absent from the culture medium or present in considerably reduced quantities. This provided a procedure for diagnosing lipidoses, by analyzing the lissamine rhodamine sphingomyelin content in the culture medium, while maintaining the cells intact.
用荧光探针丽丝胺罗丹明标记的糖鞘脂被给予培养中的皮肤成纤维细胞，并在完整细胞中水解为相应的丽丝胺罗丹明神经酰胺（N-酰基鞘氨醇）。这种荧光神经酰胺在完整细胞中转化为相应的鞘磷脂，后者被分泌到培养基中。相比之下，由于溶酶体糖脂水解酶缺乏，脂质贮积病患者来源的细胞中不会形成神经酰胺。因此，培养基中不存在荧光鞘磷脂或其含量大幅降低。这提供了一种通过分析培养基中丽丝胺罗丹明鞘磷脂含量来诊断脂质贮积病的方法，同时保持细胞完整。
