Copyright © 2006, European Society of Cardiology
Doxazosin induces activation of GADD153 and cleavage of focal adhesion kinase in cardiomyocytes en route to apoptosis
Unidad de Investigación del Servicio de Cardiología, Hospital Clínico Universitario, Santiago de Compostela, Spain
* Corresponding author. Laboratorio de Investigación Número 1, Área de Investigación y Docencia, Hospital Clínico Universitario de Santiago de Compostela, Travesía Choupana s/n, 15706 Santiago de Compostela, Spain. Tel.: +34 981 950902; fax: +34 981 950757. Email address: frlago{at}usc.es
Objective The 
1-adrenoreceptor blocker doxazosin, which in the ALLHAT trial was associated with a greater risk of heart failure than the diuretic chlorthalidone, induces the apoptosis of human and murine cardiomyocytes regardless of 1-adrenoreceptor blockade. We aimed to throw light on the mechanism of this process.
Methods Murine cardiomyocytes (HL-1) and primary cultures of human and neonatal rat cardiomyocytes were treated with 25 µmol/L doxazosin for between 0.5 and 48 h. cDNA microarray analysis, real-time RT-PCR, and Western blotting were performed to detect possible changes in gene expression and/or activation of proteins that could be involved in doxazosin-induced apoptosis.
Results Microarray analysis revealed changes in the expression of genes directly involved in the apoptotic end-stage of the cellular response to endoplasmic reticulum (ER) stress. Doxazosin considerably increased transcription and translation of gadd153, C/epbβ, and DOC-1 in cardiomyocytes as well as translocation of GADD153 to the nucleus, phosphorylation of p38 MAPK (a GADD153 activator), and the initial phosphorylation and subsequent cleavage of focal adhesion kinase (FAK). Experiments repeated following blockade of 1-adrenoreceptors showed no alteration of the above effects of doxazosin.
Conclusion Doxazosin induces the apoptosis of cardiomyocytes via the ER pathway, with increased production of C/EBPβ, GADD153 and DOC-1. Likewise it increases phosphorylation of the GADD153 activator p38 MAPK and induces first the phosphorylation, and then the cleavage, of FAK. These effects are not mediated by 1-adrenoreceptors.
KEYWORDS Apoptosis; Antihypertensive agents; Gene array analysis; Cell culture; Protein kinases