New advances in the protective mechanisms of acidic pH after ischemia: Participation of NO.

BACKGROUND: It has been previously demonstrated that the maintenance of ischemic acidic pH or the delay of intracellular pH recovery at the onset of reperfusion decreases ischemic-induced cardiomyocyte death. OBJECTIVE: To examine the role played by nitric oxide synthase (NOS)/NO-dependent pathways in the effects of acidic reperfusion in a regional ischemia model. METHODS: Isolated rat hearts perfused by Langendorff technique were submitted to 40 min of left coronary artery occlusion followed by 60 min of reperfusion (IC). A group of hearts received an acid solution (pH = 6.4) during the first 2 min of reperfusion (AR) in absence or in presence of l-NAME (NOS inhibitor). Infarct size (IS) and myocardial function were determined. In cardiac homogenates, the expression of P-Akt, P-endothelial and inducible isoforms of NOS (P-eNOS and iNOS) and the level of 3-nitrotyrosine were measured. In isolated cardiomyocytes, the intracellular NO production was assessed by confocal microscopy, under control and acidic conditions. Mitochondrial swelling after Ca2+ addition and mitochondrial membrane potential (Δψ) were also determined under control and acidosis. RESULTS: AR decreased IS, improved postischemic myocardial function recovery, increased P-Akt and P-eNOS, and decreased iNOS and 3-nitrotyrosine. NO production increased while mitochondrial swelling and Δψ decreased in acidic conditions. l-NAME prevented the beneficial effects of AR. CONCLUSIONS: Our data strongly supports that a brief acidic reperfusion protects the myocardium against the ischemia-reperfusion injury through eNOS/NO-dependent pathways.

Cyclosporine-A mimicked the ischemic pre- and postconditioning-mediated cardioprotection in hypertensive rats: Role of PKCε.

Our aim was to assess the action of cyclosporine-A (CsA) against reperfusion injury in spontaneously hypertensive rats (SHR) compared to the effects of ischemic pre- (IP) and postconditioning (IPC), examining the role played by PKCε. Isolated hearts were submitted to the following protocols: IC: 45 min global ischemia (GI) and 1h reperfusion (R); IP: a cycle of 5 min GI and 10 min of R prior to 45 min-GI; and IPC: three cycles of 30s-GI/30s-R at the start of R. Other hearts of the IC, IP and IPC groups received CsA (mitochondrial permeability transition pore inhibitor) or chelerythrine (Che, non-selective PKC inhibitor). Infarct size (IS) was assessed. TBARS and reduced glutathione (GSH) content - as parameters of oxidative damage, the expression of P-Akt, P-GSK-3ß, P-PKCε and cytochrome c (Cyc) release - as an index of mitochondrial permeability and the response of isolated mitochondria to Ca(2+) were also measured. IS similarly decreased in preconditioned, postconditioned and CsA treated heart showing the highest values in the combinations IP+CsA and IPC+CsA. TBARS decreased and GSH was partially preserved after all interventions. The content of P-Akt, P-GSK-3ß and P-PKCε increased in cytosol and decreased in mitochondria after IP and IPC. In CsA treated hearts these enzymes increased in both fractions reaching the highest values. Cyc release was attenuated and the response of mitochondria to Ca(2+) was improved by the interventions. The beneficial effects of IP and IPC were annulled when PKC was inhibited with Che. A PKCε/VDAC association was also detected. These data show that, in SHR, the CsA treatment mimicked and reinforced the cardioprotective action afforded by IP and IPC in which PKCε-mediated attenuation of mitochondrial permeability appears as the main mechanism involved.

Beneficial Consequences of One-Month Oral Treatment with Cannabis Oil on Cardiac Hypertrophy and the Mitochondrial Pool in Spontaneously Hypertensive Rats.

Introduction: It has been demonstrated the dysregulation of the cardiac endocannabinoid system in cardiovascular diseases. Thus, the modulation of this system through the administration of phytocannabinoids present in medicinal cannabis oil (CO) emerges as a promising therapeutic approach. Furthermore, phytocannabinoids exhibit potent antioxidant properties, making them highly desirable in the treatment of cardiac pathologies, such as hypertension-induced cardiac hypertrophy (CH). Objective: To evaluate the effect of CO treatment on hypertrophy and mitochondrial status in spontaneously hypertensive rat (SHR) hearts. Methods: Three-month-old male SHR were randomly assigned to CO or olive oil (vehicle) oral treatment for 1 month. We evaluated cardiac mass and histology, mitochondrial dynamics, membrane potential, area and density, myocardial reactive oxygen species (ROS) production, superoxide dismutase (SOD), and citrate synthase (CS) activity and expression. Data are presented as mean ± SEM (n) and compared by t-test, or two-way ANOVA and Bonferroni post hoc test were used as appropriate. p < 0.05 was considered statistically significant. Results: CH was reduced by CO treatment, as indicated by the left ventricular weight/tibia length ratio, left ventricular mass index, myocyte cross-sectional area, and left ventricle collagen volume fraction. The ejection fraction was preserved in the CO-treated group despite the persistence of elevated systolic blood pressure and the reduction in CH. Mitochondrial membrane potential was improved and mitochondrial biogenesis, dynamics, area, and density were all increased by treatment. Moreover, the activity and expression of the CS were enhanced by treatment, whereas ROS production was decreased and the antioxidant activity of SOD increased by CO administration. Conclusion: Based on the mentioned results, we propose that 1-month oral treatment with CO is effective to reduce hypertrophy, improve the mitochondrial pool and increase the antioxidant capacity in SHR hearts.

Non-enzymatic lipid peroxidation of microsomes and mitochondria from liver, heart and brain of the bird Lonchura striata: relationship with fatty acid composition.

The aim of this study was to examine the fatty acid composition and non-enzymatic lipid peroxidation (LP) of mitochondria and microsomes obtained from liver, heart and brain of Lonchura striata. The percentage of total unsaturated fatty acid was approximately 30-60% in the organelles from all tissues studied. Brain mitochondria and both organelles of liver exhibited the highest percentage of polyunsaturated fatty acid (PUFA) (30 and 18%, respectively). The arachidonic acid (AA) content was 7% in mitochondria of liver and brain and 3% in heart mitochondria. The percentage of docosahexanoic acid (DHA) was 8% in brain mitochondria and approximately 2-3% in heart and liver mitochondria. The peroxidizability index (PI) of brain mitochondria and both organelles from liver was higher than that of organelles from heart and brain microsomes. Liver organelles and brain mitochondria were affected by LP, as indicated by the increase in chemiluminescence and a decrease of AA and DHA. These changes were not observed during LP of brain microsomes and both organelles from heart. These results indicate: 1) PI positively correlates with PUFA percentage and LP; 2) The resistance to LP detected in heart organelles would contribute to the cardiac protection against oxidative damage.

A low degree of fatty acid unsaturation leads to high resistance to lipid peroxidation in mitochondria and microsomes of different organs of quail (Coturnix coturnix japonica).

Birds -- particularly long-lived species -- have special adaptations for preventing tissue damage caused by reactive oxygen species. The objective of the present study was to analyse the fatty acid composition and non-enzymatic lipid peroxidation of mitochondria and microsomes obtained from liver, heart and brain of quail (Coturnix coturnix japonica), a short-lived bird. Fatty acids located in total lipids of rat liver, heart and brain mitochondria and microsomes were determined using gas chromatography and lipid peroxidation was evaluated using a chemiluminescence assay. The unsaturated fatty acid content found in mitochondria and microsomes of all tissue examined was approximately 50 and 40%, respectively with a prevalence of C18:1 n9. The C18:2 n6 content in brain mitochondria was significantly lower as compared to liver and heart mitochondria. Whereas the C20:4 n6 content in mitochondria from all tissues examined and brain microsomes was approximately 6%, liver and heart microsomes exhibited lower values. C22:6 n3 was absent in liver mitochondria, very low content in liver microsomes and heart organelles (between 0.5 and 1%) and high content in brain organelles, with mitochondria having the highest value (11%). Whereas liver and heart organelles were not affected when subjected to lipid peroxidation, brain mitochondria were highly affected, as indicated by the increase in chemiluminescence and a considerable decrease of C20:4 n6 and C22:6 n3. These results indicate that a low degree of fatty acid unsaturation in liver and heart organelles of quail, a short-lived bird, may confer advantage by decreasing their sensitivity to lipid peroxidation process.

Acciones cardiovasculares comparativas de la nifedipina en perros anestesiados / Comparative cardiovascular effects of nifedipine in anesthetized dogs

En experimentos "in vitro" realizados en nuestro laboratorio se demostró una acción vasodilatadora selectiva de la nifedipina sobre el músculo liso coronario, pero aquí se usaron arterias de conductancia cuya contribución a la resistencia coronaria es despreciable. Con el objeto de detectar la selectividad de la nifedipina sobre el árbol coronario de resistencia, realizamos experimentos en perros anestesiados. En un primer grupo se realizó una curva dosis-respuesta a la nifepina y en los demás grupos cada animal recibió una única dosis de la droga. Estas dosis fueron: 0.5, 1, 5 y 10 microng x kg-1 x m-1. Se midieron los siguientes parámetros: presión arterial sistólica, diastólica (PD) y media (PM); flujo coronario sistólico, diastólico (FD) y medio a través de un medidor de flujo electromagnético colocado en la arteria circunfleja; tensión desarrollada por un segmento isométrico del ventrículo izquierdo (TD)k, frecuencia cardíaca (FC) y volumen minuto (VM) por termodilución. Con PD y FD se calculó resistencia coronaria diastólica (RCD) y con POM y VM resistencia periférica (RP). Con las dosis menores (0.5,1) la PD, PM, TD y FC no se modificaron. El FD aumentó 28 ñ 7% (p<0.05) con 0.5 y 49 ñ 11% (p<0.05) con 1. El VM aumentó 13 ñ 3% (P<0.05) y 20 ñ 6% (p<0.05) respectivamente. La RCD y RP disminuyeron 49 ñ 5% (p<0.05) y 43 ñ 5% (p<0.05) respectivamente. PD disminuyó 31 ñ 2% (p<0.05) y PM disminuyó 26 ñ 4% (p<0.05). TD disminuyó 14 ñ 5% (P<0.05). Con 10 microng x kg**1 x m-1 FD y VM...