Nakazawa M, Uehara T, Nomura Y
Department of Pharmacology, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
J Neurochem. 1997 Jun;68(6):2493-9. doi: 10.1046/j.1471-4159.1997.68062493.x.
We examined nitric oxide (NO)-induced cell death in NG108-15 cells using NO donors. Both sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine caused lactate dehydrogenase (LDH) leakage from NG108-15 cells. NO is known to increase the amount of radioisotopic labeled glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the presence of [32P]NAD and to inhibit the enzyme activity. To clarify the relationship between the NO-induced inhibition of GAPDH activity and cell death, we studied the effect of koningic acid (KA), a potent selective inhibitor of GAPDH. Both SNP and KA elicited LDH leakage, chromosomal condensation, and fragmentation of nuclei in NG108-15 cells. Gel electrophoretic analysis of cellular DNA extracted from SNP- and KA-treated cells revealed the internucleosomal DNA fragmentation typical of apoptosis in these cultures. The results suggested that in NG108-15 cells, (a) the inhibition of GAPDH activity results in apoptosis and (b) SNP-induced cell death is partly due to the NO-induced inhibition of GAPDH, perhaps by stimulating the binding of NAD to GAPDH.
我们使用一氧化氮（NO）供体检测了NG108 - 15细胞中NO诱导的细胞死亡。硝普钠（SNP）和S - 亚硝基 - N - 乙酰青霉胺均可导致NG108 - 15细胞中乳酸脱氢酶（LDH）泄漏。已知在[32P]NAD存在的情况下，NO会增加放射性同位素标记的甘油醛 - 3 - 磷酸脱氢酶（GAPDH）的量，并抑制该酶的活性。为了阐明NO诱导的GAPDH活性抑制与细胞死亡之间的关系，我们研究了GAPDH的强效选择性抑制剂 koningic acid（KA）的作用。SNP和KA均可引起NG108 - 15细胞中的LDH泄漏、染色体浓缩和细胞核碎片化。对从SNP和KA处理的细胞中提取的细胞DNA进行凝胶电泳分析，揭示了这些培养物中典型的凋亡细胞内DNA片段化。结果表明，在NG108 - 15细胞中，（a）GAPDH活性的抑制导致细胞凋亡，（b）SNP诱导的细胞死亡部分归因于NO诱导的GAPDH抑制，可能是通过刺激NAD与GAPDH的结合。
