CIC   05421
CENTRO DE INVESTIGACIONES CARDIOVASCULARES "DR. HORACIO EUGENIO CINGOLANI"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
CaMKII Activation is Involved in Angiotensin II-Mediated Cell Death
Autor/es:
JULIETA PALOMEQUE; CARLOS A VALVERDE; MARGARITA A SALAS; ALICIA MATTIAZZI
Lugar:
Orlando, FL, USA.
Reunión:
Congreso; American Heart Association (AHA). Scientific Sessions 2007; 2007
Resumen:
Angiotensin II (Ang II) is involved in the excitation contraction coupling through its modulation of Ca2+ i homeostasis via PKC. Moreover, Ang II-induced myocyte apoptosis has been described in the late 1990s. Furthermore, Ca2+-calmodulin-dependent protein kinase II (CaMKII), although typically activated by Ca2+ and calmodulin binding, has been described to be also activated by Ca2+-independent pathways that involve oxidative stress and PKC. It is also known that CaMKII initiates an apoptotic pathway in cardiac cells. Interestingly, a link between Ang II and CaMKII has never been examined. Therefore the present study was conducted to establish this nexus. To this purpose we cultivated adult cat cardiac myocytes in control conditions, treated 24 h with 1 µM Ang II and with 1µM Ang II plus 1µM KN-93, a specific CaMKII inhibitor. We evaluated cell viability and CaMKII activation by western blot of P-CaMKII and P-Thr17 of phospholamban, a specific substrate of active CaMKII. Additionally, in situ DNA nick end labelling (TUNEL) assay was used to determine apoptosis. Ang II treatment induced cell death in 64.6±3.6% versus 29.4±3.3% (control). The mortality induced by Ang II was abolished by concomitant treatment of the myocytes with the CaMKII inhibitor. The apoptotic assay followed the same pattern, being the TUNEL positive cells significantly greater in Ang II treated cells than in the group treated with Ang II and KN-93. The Ang II-induced cell death was associated with 32.6±5.6% increase in P-CaMKII and a concomitant increase in P-Thr17 was also detected (Figure). These results indicate a previously unrecognized pathway of cardiac cell death triggered by Ang II mediated by CaMKII.2+ i homeostasis via PKC. Moreover, Ang II-induced myocyte apoptosis has been described in the late 1990s. Furthermore, Ca2+-calmodulin-dependent protein kinase II (CaMKII), although typically activated by Ca2+ and calmodulin binding, has been described to be also activated by Ca2+-independent pathways that involve oxidative stress and PKC. It is also known that CaMKII initiates an apoptotic pathway in cardiac cells. Interestingly, a link between Ang II and CaMKII has never been examined. Therefore the present study was conducted to establish this nexus. To this purpose we cultivated adult cat cardiac myocytes in control conditions, treated 24 h with 1 µM Ang II and with 1µM Ang II plus 1µM KN-93, a specific CaMKII inhibitor. We evaluated cell viability and CaMKII activation by western blot of P-CaMKII and P-Thr17 of phospholamban, a specific substrate of active CaMKII. Additionally, in situ DNA nick end labelling (TUNEL) assay was used to determine apoptosis. Ang II treatment induced cell death in 64.6±3.6% versus 29.4±3.3% (control). The mortality induced by Ang II was abolished by concomitant treatment of the myocytes with the CaMKII inhibitor. The apoptotic assay followed the same pattern, being the TUNEL positive cells significantly greater in Ang II treated cells than in the group treated with Ang II and KN-93. The Ang II-induced cell death was associated with 32.6±5.6% increase in P-CaMKII and a concomitant increase in P-Thr17 was also detected (Figure). These results indicate a previously unrecognized pathway of cardiac cell death triggered by Ang II mediated by CaMKII.homeostasis via PKC. Moreover, Ang II-induced myocyte apoptosis has been described in the late 1990s. Furthermore, Ca2+-calmodulin-dependent protein kinase II (CaMKII), although typically activated by Ca2+ and calmodulin binding, has been described to be also activated by Ca2+-independent pathways that involve oxidative stress and PKC. It is also known that CaMKII initiates an apoptotic pathway in cardiac cells. Interestingly, a link between Ang II and CaMKII has never been examined. Therefore the present study was conducted to establish this nexus. To this purpose we cultivated adult cat cardiac myocytes in control conditions, treated 24 h with 1 µM Ang II and with 1µM Ang II plus 1µM KN-93, a specific CaMKII inhibitor. We evaluated cell viability and CaMKII activation by western blot of P-CaMKII and P-Thr17 of phospholamban, a specific substrate of active CaMKII. Additionally, in situ DNA nick end labelling (TUNEL) assay was used to determine apoptosis. Ang II treatment induced cell death in 64.6±3.6% versus 29.4±3.3% (control). The mortality induced by Ang II was abolished by concomitant treatment of the myocytes with the CaMKII inhibitor. The apoptotic assay followed the same pattern, being the TUNEL positive cells significantly greater in Ang II treated cells than in the group treated with Ang II and KN-93. The Ang II-induced cell death was associated with 32.6±5.6% increase in P-CaMKII and a concomitant increase in P-Thr17 was also detected (Figure). These results indicate a previously unrecognized pathway of cardiac cell death triggered by Ang II mediated by CaMKII.2+-calmodulin-dependent protein kinase II (CaMKII), although typically activated by Ca2+ and calmodulin binding, has been described to be also activated by Ca2+-independent pathways that involve oxidative stress and PKC. It is also known that CaMKII initiates an apoptotic pathway in cardiac cells. Interestingly, a link between Ang II and CaMKII has never been examined. Therefore the present study was conducted to establish this nexus. To this purpose we cultivated adult cat cardiac myocytes in control conditions, treated 24 h with 1 µM Ang II and with 1µM Ang II plus 1µM KN-93, a specific CaMKII inhibitor. We evaluated cell viability and CaMKII activation by western blot of P-CaMKII and P-Thr17 of phospholamban, a specific substrate of active CaMKII. Additionally, in situ DNA nick end labelling (TUNEL) assay was used to determine apoptosis. Ang II treatment induced cell death in 64.6±3.6% versus 29.4±3.3% (control). The mortality induced by Ang II was abolished by concomitant treatment of the myocytes with the CaMKII inhibitor. The apoptotic assay followed the same pattern, being the TUNEL positive cells significantly greater in Ang II treated cells than in the group treated with Ang II and KN-93. The Ang II-induced cell death was associated with 32.6±5.6% increase in P-CaMKII and a concomitant increase in P-Thr17 was also detected (Figure). These results indicate a previously unrecognized pathway of cardiac cell death triggered by Ang II mediated by CaMKII.2+ and calmodulin binding, has been described to be also activated by Ca2+-independent pathways that involve oxidative stress and PKC. It is also known that CaMKII initiates an apoptotic pathway in cardiac cells. Interestingly, a link between Ang II and CaMKII has never been examined. Therefore the present study was conducted to establish this nexus. To this purpose we cultivated adult cat cardiac myocytes in control conditions, treated 24 h with 1 µM Ang II and with 1µM Ang II plus 1µM KN-93, a specific CaMKII inhibitor. We evaluated cell viability and CaMKII activation by western blot of P-CaMKII and P-Thr17 of phospholamban, a specific substrate of active CaMKII. Additionally, in situ DNA nick end labelling (TUNEL) assay was used to determine apoptosis. Ang II treatment induced cell death in 64.6±3.6% versus 29.4±3.3% (control). The mortality induced by Ang II was abolished by concomitant treatment of the myocytes with the CaMKII inhibitor. The apoptotic assay followed the same pattern, being the TUNEL positive cells significantly greater in Ang II treated cells than in the group treated with Ang II and KN-93. The Ang II-induced cell death was associated with 32.6±5.6% increase in P-CaMKII and a concomitant increase in P-Thr17 was also detected (Figure). These results indicate a previously unrecognized pathway of cardiac cell death triggered by Ang II mediated by CaMKII.2+-independent pathways that involve oxidative stress and PKC. It is also known that CaMKII initiates an apoptotic pathway in cardiac cells. Interestingly, a link between Ang II and CaMKII has never been examined. Therefore the present study was conducted to establish this nexus. To this purpose we cultivated adult cat cardiac myocytes in control conditions, treated 24 h with 1 µM Ang II and with 1µM Ang II plus 1µM KN-93, a specific CaMKII inhibitor. We evaluated cell viability and CaMKII activation by western blot of P-CaMKII and P-Thr17 of phospholamban, a specific substrate of active CaMKII. Additionally, in situ DNA nick end labelling (TUNEL) assay was used to determine apoptosis. Ang II treatment induced cell death in 64.6±3.6% versus 29.4±3.3% (control). The mortality induced by Ang II was abolished by concomitant treatment of the myocytes with the CaMKII inhibitor. The apoptotic assay followed the same pattern, being the TUNEL positive cells significantly greater in Ang II treated cells than in the group treated with Ang II and KN-93. The Ang II-induced cell death was associated with 32.6±5.6% increase in P-CaMKII and a concomitant increase in P-Thr17 was also detected (Figure). These results indicate a previously unrecognized pathway of cardiac cell death triggered by Ang II mediated by CaMKII.
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