RESUMEN
We have utilized an in situ rat coronary ligation model to establish a PKC-epsilon cytochrome oxidase subunit IV (COIV) coimmunoprecipitation in myocardium exposed to ischemic preconditioning (PC). Ischemia-reperfusion (I/R) damage and PC protection were confirmed using tetrazolium-based staining methods and serum levels of cardiac troponin I. Homogenates prepared from the regions at risk (RAR) and not at risk (RNAR) for I/R injury were fractionated into cell-soluble (S), 600 g low-speed centrifugation (L), Percoll/Optiprep density gradient-purified mitochondrial (M), and 100,000 g particulate (P) fractions. COIV immunoreactivity and cytochrome-c oxidase activity measurements estimated the percentages of cellular mitochondria in S, L, M, and P fractions to be 0, 55, 29, and 16%, respectively. We observed 18, 3, and 3% of PKC-delta, -epsilon, and -zeta isozymes in the M fraction under basal conditions. Following PC, we observed a 61% increase in PKC-epsilon levels in the RAR M fraction compared with the RNAR M fraction. In RAR mitochondria, we also observed a 2.8-fold increase in PKC-epsilon serine 729 phosphoimmunoreactivity (autophosphorylation), indicating the presence of activated PKC-epsilon in mitochondria following PC. PC administered before prolonged I/R induced a 1.9-fold increase in the coimmunoprecipitation of COIV, with anti-PKC-epsilon antisera and a twofold enhancement of cytochrome-c oxidase activity. Our results suggest that PKC-epsilon may interact with COIV as a component of the cardioprotection in PC. Induction of this interaction may provide a novel therapeutic target for protecting the heart from I/R damage.
Asunto(s)
Complejo IV de Transporte de Electrones/metabolismo , Inmunoprecipitación , Precondicionamiento Isquémico , Mitocondrias Cardíacas/enzimología , Infarto del Miocardio/terapia , Daño por Reperfusión Miocárdica/prevención & control , Miocardio/enzimología , Proteína Quinasa C-epsilon/metabolismo , Animales , Vasos Coronarios/cirugía , Modelos Animales de Enfermedad , Activación Enzimática , Isoenzimas/metabolismo , Ligadura , Complejos Multiproteicos/metabolismo , Infarto del Miocardio/complicaciones , Infarto del Miocardio/enzimología , Infarto del Miocardio/patología , Daño por Reperfusión Miocárdica/enzimología , Daño por Reperfusión Miocárdica/etiología , Daño por Reperfusión Miocárdica/patología , Miocardio/patología , Fosforilación Oxidativa , Ratas , Ratas Sprague-Dawley , Troponina I/sangreRESUMEN
The title compound, C(16)H(20)N(2)O(4)S, exhibits a V-shape structure with a dihedral angle of 77.5â (11)° formed by the two benzenel rings. The mol-ecular packing is stabilized by intra-molecular and inter-molecular hydrogen bonds as well as π-π [3.738â (3)â Å] and C-Hâ¯π inter-actions.
RESUMEN
The title compound, C(20)H(13)ClN(4)O(2), was synthesized by the condensation of 4-nitro-benzohydrazide and N-(4-chlorophen-yl)-benzimidoyl chloride in N,N-dimethyl-acetamide. The asymmetric unit contains two independent mol-ecules. In one molecule, the triazole ring is oriented at dihedral angles of 23.1â (5), 85.4â (1) and 10.5â (1)° with respect to the phenyl, chlorophenyl and nitrophenyl rings, respectively. In the other molecule, the corresponding dihedral angles are 29.8â (9), 73.4â (7) and 16.4â (3)°.