Mechanism of nitric oxide‐induced apoptosis in human neuroblastoma SH‐SY5Y cells

FEBS Letters

Nomura, Yasuyuki¹  Uehara, Takashi¹  Moriya, Ryuichi¹ 

[1] Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812, Japan

关键词: Nitric oxide;    Apoptosis;    Mitochondrion;    Caspase;    DEVD-AFC;    Asp-Glu-Val-Asp-7-amino-4-trifluoromethylcoumarin;    GAPDH;    glyceraldehyde 3-phosphate dehydrogenase;    ICAD;    inhibitor of caspase-activated DNase;    LEHD;    Leu-Glu-His-Asp;    NO;    nitric oxide;    SDS;    sodium dodecyl sulfate;    VDVAD;    Val-Asp-Val-Ala-Asp;   

DOI  :  10.1016/S0014-5793(00)02167-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have attempted to elucidate the precise mechanism of nitric oxide (NO)-induced apoptotic neuronal cell death. Enzymatic cleavages of DEVD-AFC, VDVAD-AFC, and LEHD-AFC (specific substrates for caspase-3-like protease (caspase-3 and -7), caspase-2, and caspase-9, respectively) were observed by treatment with NO. Western blot analysis showed that pro-forms of caspase-2, -3, -6, and -7 are decreased during apoptosis. Interestingly, Ac-DEVD-CHO, a caspase-3-like protease inhibitor, blocked not only the decreases in caspase-2 and -7, but also the formation of p17 from p20 in caspase-3 induced by NO, suggesting that caspase-3 exists upstream of caspase-2 and -7. Bongkrekic acid, a potent inhibitor of mitochondrial permeability transition, specifically blocked both the loss of mitochondrial membrane potential and subsequent DNA fragmentation in response to NO. Thus, NO results in neuronal apoptosis through the sequential loss of mitochondrial membrane potential, caspase activation, and degradation of inhibitor of caspase-activated DNase (CAD) (CAD activation).

【 授权许可】

Unknown