Toda K, Kobayashi T, Goto I, Ohno K, Eto Y, Inui K, Okada S
Department of Neurology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
J Neurochem. 1990 Nov;55(5):1585-91. doi: 10.1111/j.1471-4159.1990.tb04942.x.
We describe here a sensitive assay method for lysosulfatide (sulfogalactosylsphingosine) in human tissues using HPLC. The method involves extraction of lipids, saponification, isolation using a C18 Sep-Pak column, derivatization with o-phthalaldehyde, and detection of the fluorescent lysosulfatide using HPLC. In control subjects, a small amount of lysosulfatide was detected in the cerebral white matter (9-35 pmol/mg of protein), spinal cord (35 pmol/mg of protein), sciatic nerve (14 pmol/mg of protein), and kidney (approximately 2 pmol/mg of protein) but not in the cerebral gray matter and liver. A marked accumulation of the lipid was noted in tissues from six patients with metachromatic leukodystrophy (MLD). The concentration of lysosulfatide was high in the cerebral white matter, spinal cord, and sciatic nerve (223-1,172 pmol/mg of protein). Even in the cerebral gray matter, kidney, and liver, where lysosulfatide was hardly detected in the control sample, a considerable amount (3-45 pmol/mg of protein) accumulated in MLD patients. The concentration and distribution pattern of lysosulfatide were similar to those of galactosylsphingosine (psychosine) accumulated in patients with Krabbe disease. Therefore, the accumulation of lysosulfatide may explain the demyelination in patients with MLD, as is the case with Krabbe disease.
我们在此描述一种使用高效液相色谱法（HPLC）检测人体组织中溶血硫酸脑苷脂（硫酸半乳糖鞘氨醇）的灵敏检测方法。该方法包括脂质提取、皂化、使用C18 Sep-Pak柱进行分离、用邻苯二甲醛衍生化，以及使用HPLC检测荧光溶血硫酸脑苷脂。在对照受试者中，在脑白质（9 - 35 pmol/毫克蛋白质）、脊髓（35 pmol/毫克蛋白质）、坐骨神经（14 pmol/毫克蛋白质）和肾脏（约2 pmol/毫克蛋白质）中检测到少量溶血硫酸脑苷脂，但在脑灰质和肝脏中未检测到。在六名异染性脑白质营养不良（MLD）患者的组织中发现该脂质有明显蓄积。溶血硫酸脑苷脂在脑白质、脊髓和坐骨神经中的浓度较高（223 - 1172 pmol/毫克蛋白质）。即使在对照样本中几乎未检测到溶血硫酸脑苷脂的脑灰质、肾脏和肝脏中，MLD患者中也蓄积了相当数量（3 - 45 pmol/毫克蛋白质）。溶血硫酸脑苷脂的浓度和分布模式与克拉伯病（Krabbe disease）患者体内蓄积的半乳糖鞘氨醇（精神鞘氨醇）相似。因此，溶血硫酸脑苷脂的蓄积可能解释了MLD患者的脱髓鞘现象，就像克拉伯病一样。
