Differential effects of iodoacetamide and iodoacetate on glycolysis and glutathione metabolism of cultured astrocytes.
Differential effects of iodoacetamide and iodoacetate on glycolysis and glutathione metabolism of cultured astrocytes.
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DOI：
10.3389/neuro.14.001.2009
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发表时间：
2009-01-01
期刊：
影响因子：
--
通讯作者：
Dringen, Ralf
中科院分区：
文献类型：
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作者：
Schmidt, Maike M;Dringen, Ralf
碘乙酰胺（IAA）和碘乙酸（IA）经常用于抑制糖酵解，因为这些化合物因其不可逆地抑制糖酵解酶甘油醛-3-磷酸脱氢酶（GAPDH）的能力而闻名。然而，用这样的硫醇试剂处理对脑细胞的谷胱甘肽（GSH）代谢的后果尚未被探索。暴露的星形胶质细胞丰富的原代培养物IAA或IA的浓度高达1毫米剥夺了细胞的GSH，抑制细胞GAPDH活性，降低细胞乳酸的产生，并导致延迟的细胞死亡，是可检测的孵育90分钟后。然而，这两种巯基试剂在剥夺细胞GSH和抑制星形胶质细胞糖酵解的潜力方面存在很大差异。IAA比IA更有效地耗尽细胞GSH含量，如在约10和100 μ M的浓度下分别观察到的IAA和IA的半数最大效应所证明的。相比之下，IA在灭活GAPDH和乳酸产生方面是高效的，在低于100 μ M的浓度下已经观察到半最大效应，而IAA必须以10倍高的浓度施用以抑制50%的乳酸产生。IAA和IA影响培养的星形胶质细胞的GSH含量和糖酵解的这些实质性差异表明，为了抑制星形胶质细胞糖酵解而不实质性损害细胞GSH代谢，IA -而不是IAA -应以低浓度和/或短孵育期使用。
Iodoacetamide (IAA) and iodoacetate (IA) have frequently been used to inhibit glycolysis, since these compounds are known for their ability to irreversibly inhibit the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). However, the consequences of a treatment with such thiol reagents on the glutathione (GSH) metabolism of brain cells have not been explored. Exposure of astroglia-rich primary cultures to IAA or IA in concentrations of up to 1 mM deprived the cells of GSH, inhibited cellular GAPDH activity, lowered cellular lactate production and caused a delayed cell death that was detectable after 90 min of incubation. However, the two thiol reagents differed substantially in their potential to deprive cellular GSH and to inhibit astrocytic glycolysis. IAA depleted the cellular GSH content more efficiently than IA as demonstrated by half-maximal effects for IAA and IA that were observed at concentrations of about 10 and 100 muM, respectively. In contrast, IA was highly efficient in inactivating GAPDH and lactate production with half-maximal effects observed already at a concentration below 100 muM, whereas IAA had to be applied in 10 times higher concentration to inhibit lactate production by 50%. These substantial differences of IAA and IA to affect GSH content and glycolysis of cultured astrocytes suggest that in order to inhibit astrocytic glycolysis without substantially compromising the cellular GSH metabolism, IA - and not IAA - should be used in low concentrations and/or for short incubation periods.