SELECTIVE KILLING OF CELLS WITH OXIDATIVE DEFECTS IN GALACTOSE MEDIUM - A SCREENING-TEST FOR AFFECTED PATIENT FIBROBLASTS
SELECTIVE KILLING OF CELLS WITH OXIDATIVE DEFECTS IN GALACTOSE MEDIUM - A SCREENING-TEST FOR AFFECTED PATIENT FIBROBLASTS
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DOI：
10.1007/bf01800243
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发表时间：
1992-01-01
影响因子：
4.2
通讯作者：
ROBINSON, BH
中科院分区：
文献类型：
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作者：
PETROVABENEDICT, R;BUNCIC, JR;ROBINSON, BH
几年前的研究表明，线粒体呼吸有缺陷的中国仓鼠(CH)成纤维细胞能够保持细胞内的能量电荷，并保持正常的生长速度，主要是通过将葡萄糖转化为乳酸的速率增加一倍(Sodeberg等人t980；Day和scher t982)。我们在线粒体呼吸链缺陷患者的培养皮肤纤维刺中也进行了类似的观察(Robinson等人1986,1987A)。在培养液中用半乳糖(5 mmoL/L)替代葡萄糖对氧化缺乏性的CH成纤维细胞是致命的(Sodeberg等，1980)。我们通过将皮肤成纤维细胞传代培养到含有5 mmo1/L半乳糖的~-MEM中，研究了半乳糖对不同氧化缺陷症患者多种细胞系活力的影响。所使用的细胞系如Robinson等人(1987b)、Robinson等人(1990)和Gterum等人(T988)所描述的那样。而对照组细胞的生长和存活模式与在葡萄糖中培养的细胞无明显差异，严重新生儿复合体I缺乏症、细胞色素氧化酶缺乏症(Leigh病)和多呼吸链复合体缺乏症患者在传代后24 h内无活细胞存活。重度丙酮酸脱氢酶复合体缺乏症、复合体I缺乏伴Leigh病和部分细胞色素氧化酶缺乏症患者的细胞培养2周后生长极差，细胞死亡。来自丙酮酸脱氢酶复合体部分缺乏症、丙酮酸羧酶缺乏症、肝脏特异性细胞色素氧化酶缺乏症、Leber遗传性视神经病变(mtDNA11778突变的同质)和Kearns-Sayre综合征患者的细胞株在含半乳糖的培养液中培养未见不良反应。当这些细胞系在含有半乳糖+25#摩尔/L叠氮的培养液中重新培养时，结果是相同的，只是两个Leber遗传性视神经病变细胞系中有一个生长很差并死亡。表现出对半乳糖代谢易感性的细胞系似乎是那些具有更明显或更严重氧化缺陷的细胞系，并且易感性的程度似乎与氧化缺陷的性质有关。因此，在半乳糖介质中，严重的复合体I缺乏症、多重呼吸链缺乏症和严重的细胞色素氧化酶缺乏症细胞几乎立即被杀死，而
It was shown a number of years ago that Chinese hamster (CH) fibroblasts with defects in mitochondrial respiration were able to keep up the energy charge within the celt and maintain normal growth rates, primarily by doubling the rates of conversion of glucose to lactic acid (Sodeberg et al t980; Day and Schemer t982). We have made similar observations in cultured skin fibrobtasts from patients with defects of the mitochondrial respiratory chain (Robinson et al 1986, 1987a). Substitution of galactose (5 mmol/L) for glucose in the culture medium was shown to be lethal for oxidative-deficient CH fibroblasts (Sodeberg et al 1980). We investigated the effects of galactose on the viability of a variety of cell lines from patients with different oxidative defects by subculturing the skin fibroblasts into~-MEM containing 5 mmol/L galactose. Cell lines used were as described by Robinson et al (1987b), Robinson et al (1990) and Gterum et al (t988). While control cell lines displayed a pattern of growth and survival no different from those grown in glucose medium, within 24 h of subculture no live cells survived from patients with severe neonatal complex I deficiency, cytochrome oxidase deficiency (Leigh disease), and multiple respiratory chain complex deficiency. Cells from patients with severe pyruvate dehydrogenase complex deficiency, complex I deficiency with Leigh disease, and partial cytochrome oxidase deficiency (Leigh disease) showed extremely poor growth with cell death after 2 weeks in culture. Cell lines from patients with partial deficiency of the pyruvate dehydrogenase complex, pyruvate carboxylase deficiency, liver-specific partial cytochrome oxidase deficiency, Leber hereditary optic neuropathy (homoplasmic for the mtDNA 11 778 mutation), and Kearns-Sayre syndrome showed no adverse effects of culture in galactose-containing medium. When these cell lines were reassessed by culture in a medium containing galactose plus 25# mol/L azide, the results were the same except that one of two cell lines tested with Leber hereditary optic neuropathy (LHON) grew very poorly and died. The cell lines showing susceptibility to galactose metabolism appear to be those with the more overt or severe oxidative defects, and the extent of the susceptibiity appears to bear some relationship to the nature of the oxidative defect. Thus severe complex I deficiency, multiple respiratory chain deficiency and severe cytochrome oxidase deficiency cells are almost immediately killed in galactose medium, whereas