Protective properties of neoechinulin a against SIN-1-induced neuronal cell death
Protective properties of neoechinulin a against SIN-1-induced neuronal cell death
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DOI：
10.1093/jb/mvh103
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发表时间：
2004-07-01
影响因子：
2.7
通讯作者：
Arai, T
中科院分区：
文献类型：
--
作者：
Maruyama, K;Ohuchi, T;Arai, T
过氧亚硝酸盐（ONOO-）被认为参与神经退行性过程。为了筛选针对ONOO（-）诱导的细胞死亡的神经保护化合物，我们开发了基于96孔的测定程序，用于测量在由ONOO-产生剂3-（4-吗啉基）悉尼酮亚胺盐酸盐（SIN-1）和Cu/Zn超氧化物（O-2（-））歧化酶抑制剂N，N-二乙基二硫代氨基甲酸钠三水合物（DDC）引起的氧化应激下存活的细胞数。使用这些程序，我们获得了一种微生物代谢产物，该代谢产物可将原代神经元细胞从SIN-1诱导的损伤中拯救出来，但不能从DDC诱导的损伤中拯救出来。通过NMR分析，该化合物被鉴定为新棘球蛋白A，一种抑制脂质氧化的抗氧化剂化合物。我们发现，该化合物拯救神经元细胞，如原代神经元细胞和分化的PC12细胞免受细胞外ONOO-诱导的损伤。然而，非神经元细胞、未分化的PC12细胞和成纤维细胞系3Y1的细胞未被拯救。新棘球蛋白A具有清除、神经营养因子样和抗凋亡活性。该化合物特异性清除ONOO-，但不清除O-2（-）或一氧化氮（NO）。与已知的神经保护物质如神经生长因子和银杏叶提取物类似，新棘球蛋白A抑制SIN-1诱导的半胱天冬酶-3样蛋白酶的活化并增加NADH-脱氢酶活性。这些结果表明，neoechinulin A可能是有用的，以防止神经元细胞死亡的神经退行性疾病。
Peroxynitrite (ONOO-) is thought to be involved in the neurodegenerative process. To screen for neuroprotective compounds against ONOO(-)induced cell death, we developed 96-well based assay procedures for measuring surviving cell numbers under oxidative stress caused by 3-(4-morpholinyl) sydnonimine hydrochloride (SIN-1), a generator of ONOO-, and sodium NN-dietyldithiocarbamate trihydrate (DDC), an inhibitor of Cu/Zn superoxide (O-2(-)) dismutase. Using these procedures, we obtained a microbial metabolite that rescued primary neuronal cells from SIN-1-induced damage, but not from DDC-induced damage. By NMR analysis, the compound was identified as neoechinulin A, an antioxidant compound that suppresses lipid oxidation. We found that the compound rescues neuronal cells such as primary neuronal cells and differentiated PC12 cells from damage induced by extracellular ONOO-. However, non-neuronal cells, undifferentiated PC12 cells and cells of the fibroblast cell line 3Y1 were not rescued. Neoechinulin A has scavenging, neurotrophic factor-like and antiapoptotic activities. This compound specifically scavenges ONOO-, but not O-2(-) or nitric oxide (NO). Similar to known neuroprotective substances such as nerve growth factor and extracts of Gingko biloba leaves, neoechinulin A inhibits the SIN-1-induced activation of caspase-3-like proteases and increases NADH-dehydrogenase activity. These results suggest that neoechinulin A might be useful for protecting against neuronal cell death in neurodegenerative diseases.