Clinical and Molecular Genetical Research for the Treatment of Mitochondrial Encephalomyopathies

线粒体脑肌病是由线粒体内膜电子传递系统功能障碍引起的。主要的治疗方法尚未确立，部分原因是从线粒体酶活性和线粒体（mt）DNA分析的系列变化来看，治疗的临床效果得到了很好的评价。建立了血小板线粒体酶活性和线粒体DNA分析方法。我们还开发了一水肌酸（Cr-H_2 O）治疗线粒体脑肌病的方法，并对分离的PLT进行了微型化分析，用于蛋白质、复合物II+III（II+III）、复合物IV（IV）和柠檬酸合酶（CS）的测定。我们新开发了（IV）的小型化测定。应用PCR/RFLP方法检测了1例MELAS患者的线粒体DNA A3243 G突变，并对MELAS患者的脑卒中样发作及二氯醋酸钠治疗的疗效进行了评价 关于我们 e（DCA）处理，使用PLT线粒体酶测定。虽然她的肌无力在细胞色素c治疗后得到改善，但神经系统症状（包括头痛、呕吐和视觉障碍）发展为顽固性头痛，这是一种轻度的中风样发作。DCA治疗后顽固性严重头痛改善。卒中样发作时PLT（II+III）、（IV）和（IV/CS）降低，DCA治疗改善了PLT（II+III）、（IV）和（IV/CS）。对2例因线粒体DNA A3243 G突变所致的MELAS患儿，给予口服Cr-H_2 O（0.2g/kg/d，分两次服用，第一次服用2周，第二次服用0.08- 0.12g/kg/d，每日2次或3次，共4-10个月）治疗。结果表明，补充Cr-H_2 O可改善MELAS的脑功能，提示Cr-H_2 O治疗线粒体脑肌病的可能性。少

Mitochondrial encephalomyopathy is caused by mitochondrial dysfunction mainly in electron transport system in the inner mitochondrial membrane. The essential treatment has not yet been established, partly because the clinical effect of the treatment has been well evaluated from the point of the serial changes of mitochondrial enzyme activity and mitochondrial (mt) DNA analyses. We developed the methods of mitochondrial enzyme activities and mtDNA analyses in platelet (PLT). We also developed creatine monohydrate (Cr-H_2O) treatment for the mitochondrial encephalomyopathies.Miniaturized assays for isolated PLT were employed for the measurement of protein, complex II+III (II+III), complex IV (IV), and citrate synthase (CS) assays using a microplate reader. We newly developed the miniaturized assay for (IV). A3243G mutation of mtDNA was analyzed with PCR/RFLP methtod for the isolated PLT.We evaluated the stroke-like episode in a patient with MELAS and the efficacy of sodium dichloroacetat … More e (DCA) treatment using PLT mitochondrial enzyme assay. Although her muscle weakness improved with cytochrome c treatment, neurological symptoms including headache, vomiting and visual disturbance developed to intractable headache, which was a mild form of stroke-like episode. The intractable severe headache improved with DCA treatment. PLT (II+III), (IV) and (IV/CS) decreased at the stroke-like episode and DCA treatment improved PLT (II+III), (IV), and (IV/CS). Ratio of A3243G mutation of mtDNA was not changed before and after DCA treatment.Cr-H_2O was orally supplemented (0.2 g/kg/day, divided into two for the first two weeks followed by 0.08-0.12 g/kg/day twice or three times a day for 4-10 months) in two girls with MELAS due to an nt A3243G mtDNA mutation. The brain function was evaluated with frequency analysis of EEG and ^1H MRS.the results demonstrated that Cr-H_2O supplement improved the brain function of MELAS, suggesting the possibility of Cr-H_2O treatment for mitochondrial encephalomyopathy. Less