皮肤成纤维细胞Filipin染色诊断C型Niemann-Pick病

The Value of Filipin Staining of Cultured Skin Fibroblasts in Diagnosis of Niemann-Pick Disease Type C

目的建立皮肤成纤维细胞filipin染色诊断C型Niemann-Pick病(NPC)方法。方法原代培养小鼠皮肤成纤维细胞,用脂蛋白缺乏血清(LPDS)培养5 d后,与人低密度脂蛋白(LDL)共培养24 h,经filipin染色并共聚焦照相。结果纯合子小鼠(NPC-1^(-/-))小鼠组核周有密集的蓝色未酯化胆固醇聚集,而野生型小鼠(NPC一1^(+/+))核周无未酯化胆固醇聚集。结论通过filipin染色可诊断NPC病。

Objective To evaluate the significance of fitipin staining of cultured skin fibroblasts in diagnosis of Niemann-Pick disease type C（NPC）. Methods Skin fibroblasts obtained from wildtype mice and mutant mice had been cultured with lipoprotein-deficient fetal bovine serum（LPDS）, for 5 days. The fibroblasts were then cocultured with LDL for 24 hours, and subjected to filipin staining. Under a confoeal microscope, photography was harvested with excitation of violet light. Both mutant and wild-type mice were analyzed and evaluated genetically and neuropathologically. Results Filipin staining showed an intense punctate pattern of fluorescence concentrated around the nuclei of the cultured fibroblasts, consistent with the aberrant accumulation of unesterified cholesterols in cells of the NPC mice. In a sharp contrast, the filipin fluorescence was faintly distributed inside the cytoplasma of the cells in the wild-type mice. Conclusions Filipin staining could reliably diagnose the Niemann-Pick disease type C.