Roscher A, Molzer B, Bernheimer H, Stöckler S, Mutz I, Paltauf F
Pediatr Res. 1985 Sep;19(9):930-3. doi: 10.1203/00006450-198509000-00013.
The sequence of reactions involved in plasmalogen biosynthesis has been evaluated in cultured fibroblasts of patients with the cerebrohepatorenal syndrome. A double-label, double-substrate incubation using [1-14C] hexadecanol and 1-0-[9', 10'-3H]hexadecylglycerol was performed to monitor the relative rates of peroxisomal and microsomal biosynthesic steps. [14C] radioactivity associated with 1'-alkenyl groups of plasmalogens was found to be drastically reduced in fibroblasts of affected patients whereas [3H] incorporation was apparently normal. This finding is specific for cerebrohepatorenal syndrome fibroblasts since cell lines of patients with childhood adrenoleukodystrophy and neuronal ceroidlipofuscinosis utilized the lipid precursors of plasmalogen biosynthesis at normal rates. The results show that the defect in plasmalogen synthesis in the cerebro-hepato-renal syndrome is restricted to the peroxisomal steps. The finding of normal microsomal biosynthetic steps was exploited to devise a novel diagnostic assay in fibroblasts and amniocytes based on the comparison of [3H/14C] isotope ratios within aldehydes released from plasmalogens by acid hydrolysis. The procedure can be completed with a minimal amount of cells since it renders quantitative analyses unnecessary. Therefore, this technique appears ideally suited for the sensitive and safe prenatal diagnosis of the cerebro-hepato-renal syndrome.
已在脑肝肾综合征患者的培养成纤维细胞中评估了缩醛磷脂生物合成中涉及的反应序列。使用[1-¹⁴C]十六醇和1-O-[9',10'-³H]十六烷基甘油进行双标记、双底物孵育,以监测过氧化物酶体和微粒体生物合成步骤的相对速率。发现与受影响患者成纤维细胞中缩醛磷脂的1'-烯基相关的[¹⁴C]放射性大幅降低,而[³H]掺入显然正常。这一发现是脑肝肾综合征成纤维细胞所特有的,因为儿童肾上腺脑白质营养不良和神经元蜡样脂褐质沉积症患者的细胞系以正常速率利用缩醛磷脂生物合成的脂质前体。结果表明,脑肝肾综合征中缩醛磷脂合成的缺陷仅限于过氧化物酶体步骤。基于酸水解从缩醛磷脂释放的醛中[³H/¹⁴C]同位素比率的比较,利用微粒体生物合成步骤正常这一发现,在成纤维细胞和羊水中设计了一种新的诊断试验。由于该方法无需进行定量分析,因此只需使用极少量细胞即可完成该过程。因此,这项技术似乎非常适合脑肝肾综合征的灵敏且安全的产前诊断。