THE RESEARCH OF THE THERAPY FOR GAUCHER DISEASE WITH CENTRAL NERVOUS INVOLVEMENT BY THE LOW MOLECULAR COMPOUNDS.

戈谢病（GD）是最常见的鞘脂病形式，由β-葡萄糖苷酶（β-Glu）缺陷引起。碳水化合物模拟物N-辛基-β-维列胺（NOV）是β-Glu的抑制剂。当应用于培养的具有F213 I β-Glu突变的GD成纤维细胞时，NOV增加了突变酶的蛋白水平并上调了细胞酶活性。在F213 I纯合细胞中观察到NOV的最大效应，其中30 μ M NOV处理4天导致酶活性增加约6倍，高达对照细胞中活性的约80%。NOV在具有其他β-Glu突变N370 S、L444 P、84 CG和RecNciI的细胞中无效。免疫荧光和细胞分级显示F213 I突变酶在经NOV处理的细胞的溶酶体中的定位。与此一致，NOV恢复了F213 I纯合细胞中14 C标记的葡萄糖神经酰胺的清除。F213 I突变体β-Glu在体外中性pH下迅速失去活性，NOV可减弱这种pH依赖性活性丧失。这些结果表明，NOV可作为化学伴侣加速F213 I突变体β-Glu的转运和成熟，并可能表明该化合物对GD的治疗价值。

Gaucher disease(GD) is the most common form of sphingolipidosis and is caused by a defect of beta-glucosidase (beta-Glu). A carbohydrate mimic N-octyl-beta-valienamine(NOV) is an inhibitor of beta-Glu. When applied to cultured GD fibroblasts with F213I beta-Glu mutation, NOV increased the protein level of the mutant enzyme and up-regulated cellular enzyme activity. The maximum effect of NOV was observed in F213I homozygous cells in which NOV treatment at 30 microM for 4 days caused a approximately 6-fold increase in the enzyme activity, up to approximately 80% of the activity in control cells. NOV was not effective in cells with other beta-Glu mutations, N370S,L444P,84CG and RecNciI. Immunofluorescence and cell fractionation showed localization of the F213I mutant enzyme in the lysosomes of NOV-treated cells. Consistent with this, NOV restored clearance of 14C-labeled glucosylceramide in F213I homozygous cells. F213I mutant beta-Glu rapidly lost its activity at neutral pH in vitro and this pH-dependent loss of activity was attenuated by NOV. These results suggest that NOV works as a chemical chaperone to accelerate transport and maturation of F213I mutant beta-Glu and may suggest a therapeutic value of this compound for GD.