RESUMEN
The protective effects and roles of AT1-receptor antagonists (AT1-RA) or angiotensin-converting enzyme inhibitors (ACEI) on vascular endothelial cell (EC) injury during hypoxia are not entirely known. Therefore, we investigated these effects and mechanisms in human aortic (HA) EC. DNA fragmentation, Lactate dehydrogenase (LDH) release, and caspase-3 activity were measured in cultured HAEC after exposure to hypoxia in the presence or absence of an AT1-RA (candesartan, CS) and/or an ACEI (temocaprilat, TC). Next, we investigated endothelial cell nitric oxide synthase (ecNOS) and inducible (i) NOS to determine the role of the bradykinin(BK)-NO pathway in the protective effect on ACEI and AT1-RA in the setting of hypoxia-induced apoptosis. Exposure to hypoxia increased DNA fragmentation in HAEC associated with the activation of caspase-3, but did not affect LDH release. In addition, hypoxia induced ecNOS mRNA but not mRNA iNOS. CS and/or TC reduced apoptosis induced by hypoxia in a dose-dependent manner, and significantly increased BK and ecNOS expression. This effect was attenuated by the kinin B2 receptor antagonist, HOE 140, and the NOS inhibitor, N-nitro-L-arginine methylester (L-NMMA). Hypoxia activates the pathway leading to apoptosis by enhancing caspase-3 activity. Both CS and TC can ameliorate hypoxia-induced apoptosis in HAEC through inhibiting caspase-3 activation by enhancing ecNOS activity, via the accumulation of BK.
Asunto(s)
Bloqueadores del Receptor Tipo 1 de Angiotensina II , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Apoptosis/efectos de los fármacos , Bencimidazoles/farmacología , Endotelio Vascular/enzimología , Óxido Nítrico Sintasa/metabolismo , Tetrazoles/farmacología , Tiazepinas/farmacología , Aorta , Compuestos de Bifenilo , Bradiquinina/metabolismo , Caspasa 3 , Caspasas/metabolismo , Hipoxia de la Célula , Células Cultivadas , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/patología , Humanos , Imidazoles/farmacología , Necrosis , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo III , Piridinas/farmacología , omega-N-Metilarginina/farmacologíaRESUMEN
Alpha-tocopherol has been reported to exert an anti-atherogenesis effect. We attempted to clarify the effect of alpha-tocopherol-both as an antioxidant and as a nonantioxidant--on apoptosis induced by oxidized low-density lipoprotein (LDL) or oxysterols. Oxidized LDL and oxysterols induced necrosis and/or apoptosis of vascular endothelial cells. The induction of apoptosis was associated with increased caspase-3 activity and the generation of intracellular reactive oxygen species, both the effects of which were attenuated by alpha-tocopherol. Apoptosis was also decreased by beta-tocopherol or intracellular radical scavengers, but these suppressive effects were less than those of alpha-tocopherol. Neither beta-tocopherol nor the scavengers had pronounced effect on caspase-3 activity, but each of them decreased the generation of reactive oxygen species to the same extent as alpha-tocopherol. Our study suggests that alpha-Toc protects against apoptosis not only by scavenging reactive oxygen species, but also by inhibiting caspase activity, which means that its activity may exceed that of a mere antioxidant.
Asunto(s)
Antioxidantes/farmacología , Apoptosis/efectos de los fármacos , Endotelio Vascular/efectos de los fármacos , alfa-Tocoferol/farmacología , Caspasa 3 , Caspasas/metabolismo , Células Cultivadas , Relación Dosis-Respuesta a Droga , Endotelio Vascular/metabolismo , Endotelio Vascular/patología , Humanos , Hidroxicolesteroles/farmacología , Cetocolesteroles/farmacología , Lipoproteínas LDL/farmacología , Necrosis , Especies Reactivas de Oxígeno/metabolismo , Factores de TiempoRESUMEN
A total of 160 resin-simulated canal blocks with 20-degree or 30-degree curvature were prepared by ProFile rotary instruments, GT rotary files, and the balanced force technique with Flex-R files. Using an image analysis computer application, the pre- and postoperative images were stored and superimposed, and then the amount of material removed from the preoperative inner and outer curved walls was measured at five levels in the apical 5 mm of the canal. The time required for canal preparation, including irrigation, and the time taken to change instruments was recorded. At 1 mm from the apical end, the directions of canal transportation were most frequently toward the outer aspect of the curvature, with the only exception being the canals shaped by the balanced force technique in which more was removed from the inner aspect. The balanced force technique required more preparation time than the rotary instrumentation.