H9c2 cardiac myoblasts undergo apoptosis in a model of ischemia consisting of serum deprivation and hypoxia: inhibition by PMA

FEBS Letters

Bonafè, Francesca¹  Bonavita, Francesca¹  Facchini, Annalisa¹  Giordano, Emanuele¹  Columbaro, Marta²  Stefanelli, Claudio¹  Guarnieri, Carlo¹  Caldarera, Claudio Marcello¹ 

[1] Department of Biochemistry ‘G. Moruzzi’, University of Bologna, Via Irnerio, 48 40126 Bologna, Italy;Laboratory of Neuromuscular Pathology, IOR, Bologna, Italy

关键词: Ischemia;    H9c2;    Caspase;    Apoptosis;    Bcl-2 family proteins;    Phorbol-12-myristate-13-acetate;    PMA;    phorbol-12-myristate-13-acetate;    MTT;    3-[4;    5-dimethylthiazol-2-yl]-2;    5-diphenyltetrazolium bromid;    Ac-DEVD-AMC;    Ac-Asp-Glu-Val-Asp-7-amino-4-methylcoumarin;    Ac-IETD-AMC;    Ac-Ile-Glu-Thr-Asp-7-amino-4-methylcoumarin;    Ac-LEHD-AFC;    Ac-Leu-Glu-His-Asp-7-amino-4-trifluoromethylcoumarin;    z-DEVD-cmk;    benzyloxycarbonyl-Asp-Glu-Val-Asp-chloromethylketone;    JC-1;    5;    5′;    6;    6′-tetrachloro-1;    1′;    3;    3′-tetraethylbenzimidazol-carbocyanine iodide;    TUNEL;    terminal (TdT)-mediated dUTP nick end-labeling;    ΔΨm;    mitochondrial membrane potential;    PKC;    protein kinase C;    IAPs;    inhibitor of apoptosis proteins;    ARC;    apoptosis repressor with caspase recruitment domain;    DAPI;    4;    6-diamidino-2-phenylindole;   

DOI  :  10.1016/S0014-5793(03)00029-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Cardiac myocytes undergo apoptosis under condition of ischemia. Little is known, however, about the molecular pathways that mediate this response. We show that serum deprivation and hypoxia, components of ischemia in vivo, resulted in apoptosis of rat ventricular myoblast cells H9c2. Hypoxia alone did not induce significant apoptosis for at least 48 h, but largely increased the proapoptotic action of serum deprivation. H9c2 cells apoptosis is evidenced by an increase in terminal (TdT)-mediated dUTP nick end-labeling-positive nuclei and by activation of caspases 3, 6, 7 and 9, and loss of mitochondrial functions. In this model of simulated ischemia, represented by serum deprivation plus hypoxia, cardiomyoblasts apoptosis was associated with a p53-independent Bax accumulation and with a down-regulation of Bcl-xL, whereas the levels of cIAP-1, cIAP-2 and X-IAP proteins did not change. Phorbol-12-myristate-13-acetate significantly reduced the induction of apoptosis, inhibiting caspase 3 cleavage, Bax accumulation, Bcl-xL down-regulation as well as restoring cell viability.

【 授权许可】

Unknown