[Effect of ischemia and rotenone on lipid and adenine nucleotide levels and functional activity of heart mitochondria]. / Vliianie ishemii i rotenona na soderzhanie lipidov, adeninnukleotidov i funktsional'nuiu aktivnost' mitokhondrii serdtsa.

Myocardial mitochondria (MCh), isolated with rotenone (MCh + RO) and in absence of rotenone (MCh - RO), were studied in rabbits with ischemia (0.5 hr autolysis) and in controls. Content of acyl-CoA was increased by 50%, linoleic acid - by 49% and lysophosphatidyl choline - by 37% in MCh + RO of control animals as compared with the MCh - RO preparation. In the MCh + RO preparation from rabbits with ischemia content of acyl-CoA was increased by 62%, while concentration of free fatty acids (FFA) and phospholipids was similar to those of the MCh - RO preparation. After isolation of mitochondria with rotenone composition of adenine nucleotides was distinctly altered. Content of ATP was decreased by 27% in mitochondria of both ischemic and control animals, although total amount of adenine nucleotides was decreased only slightly. As shown by estimation of succinate oxidation and membrane potential rotenone did not affect the mitochondrial respiration. In mitochondria of ischemic rabbits concentrations of FFA and lysophosphatidyl choline were increased, whereas content of ATP, total amount of adenine nucleotides, the rate of succinate oxidation and membrane potential were decreased. The method developed, isolation of mitochondria with rotenone, may be used in studies of the role of acyl-CoA in energy metabolism of cells.

Hereditary polymyopathy and cardiomyopathy in the Syrian hamster. II. Development of heart necrotic changes in relation to defective mitochondrial function.

The mitochondrial oxidative phosphorylation, calcium and magnesium contents, and swelling-contraction activity were investigated in relation to the progression of the hereditary hamster cardiomyopathy. The assessment was made in animals between 22 and 232 days of age, which were divided into 7 groups according to stage of disease. In 24-day-old hamsters prior to development of heart necrotic changes, the membrane permeability of isolated mitochondria was altered. In 50-day-old animals, at a stage of disease when myocardial cells undergo degeneration, a defect of oxidative phosphorylation resulting from an increase in mitochondrial calcium was demonstrated. With culmination of the heart necrotic changes, at close to 100 days of age, mitochondrial dysfunction and calcium overload were maximal. There was a transient improvement during the healing stage, but the situation deteriorated with the occurrence of circulatory failure. Since the mitochondrial respiratory pattern and calcium overload parallel the cardiac degeneration, it is inferred that the cell energy depletion is a functional consequence of an abnormal calcium influx.

Lidocaine-induced reduction in size of experimental myocardial infarction.

In anesthetized open chest dogs, the effects of therapeutic doses of lidocaine on myocardial cell respiration, creatine kinase depletion, left ventricular stroke work and cardiac necrosis were assessed. The dogs were subjected to 40 minutes of occlusion of the left anterior descending coronary artery, followed by 5 hours of reperfusion. Group I (12 dogs) had infusion of saline solution; group II (8 dogs) had infusion of 0.2 mg/kg per min of lidocaine (serum level 16.8 +/- 1.4 microgram/ml); group III (5 dogs) had infusion of 0.04 mg/kg per min or lidocaine (serum level 3.6 micrograms/ml). Ischemic regional myocardial blood flow (measured by 9 micrometer spheres of strontium-85) was 6.34 +/- 1.62 ml/100 mg per min in group I, 1.48 +/- 0.59 in group II (p < 0.05) and 1.32 +/- 0.50 in group III (p < 0.05). Oxygen consumed during conversion of adenosine diphosphate to adenosine triphosphate in mitochondria from control and lidocaine-treated ischemic tissue was depressed (p < 0.05) and correlated (r = 0.63) with creatine kinase depletion. Left ventricular stroke work was not significantly different among the three groups. Infarct size (in percent of left ventricular weight) was 12.6 +/- 2.0 for group I, 4.8 +/- 1.2 for group II (p < 0.01) and 4.8 +/- 2.5 for group III (p < 0.05). The data suggest that the reduction of myocardial infarct size by lidocaine was not dependent on enhanced myocardial blood flow and was independent of left ventricular stroke work.

The relative time course of early changes in mitochondrial function and intracellular pH during hypoxia in the isolated toad ventricle strip.

Changes in the intracellular H+ ion concentration (pHi) and in the oxidation-reduction state of the respiratory chain were measured spectrophotometrically in the isolated ventricle strip from the toad (Bufo marinus). The relative time course of the delta pHi as indicated by changes in light absorption of the pH dye neutral red, cytochrome c, and peak isometric twitch tension were compared during transient hypoxic episodes. The first detectable change in pH occurred 4.5 minutes after the peak twitch tension began to decrease with the onset of hypoxia. The initial decrease in tension and reduction of cytochrome c occurred at a similar time prior to the change in pHi. On reoxygenation, cytochrome c rapidly became oxidized, and the pHi and tension recovered more slowly. During acidification by increasing superfusate PCO2, pHi and tension decreased together, and cytochrome c did not change significantly. Thus, although changes in pHi do affect mechanical performance, these results show that mechanical dysfunction pursuant to hypoxia is not directly attributable to intracellular acidification.

Changes in energy transduction efficiency in mitochondria isolated from ischemic myocardium.

The effects of temporary coronary artery occlusion (TCAO) in vivo and ischemia in situ (autolysis) on mitochondrial functions and inner membrane permeability were investigated. It was found that one day after TCAO the permeability of inner membrane to H+ was increased 64% and the maximal activity of the glutamate plus malate oxidase system was reduced 35% with respect to control. The proton-electrochemical gradient and the rate of oxidative phosphorylation were decreased 35% and 55%, respectively. The most significant alterations during autolysis were observed in respiratory control ratio, state IV respiration and in H+ permeability of inner membrane. The H+ permeability of inner membrane increased progressively with time, and after 4 hr of autolysis it was 378% higher than the control. The data suggest that the alterations in energy transduction efficiency of mitochondria isolated from ischemic myocardium depend largely on the increase of inner membrane permeability to H+.

Substrate effects on mitochondrial function and tissue lipids in low-flow hypoxia of isolated perfused rat hearts.

A possible causal relationship between tissue FFA contents and the depression in mitochondrial oxidative phosphorylation in myocardial ischaemia has been suggested. To test this hypothesis, the effects of different substrates added to the perfusates of hypoxic, low-flow perfused hearts were examined on oxidative phosphorylation catalysed by mitochondria isolated from such tissue. In an additional series of experiments tissue neutral glyceride and FFA levels were analysed and correlated with changes in mitochondrial function. Mitochondria isolated from hearts with a high tissue FFA content exhibited the lowest ADP/O ratios, RCI and QO2 values. On the other hand, mitochondria isolated from hearts with reduced FFA contents, performed significantly better with respect to these parameters of mitochondrial function studied.

Myocardial depression after elective ischemic arrest. Subcellular biochemistry and prevention.

The hemodynamic and cardiac biochemical effects of global ischemic arrest during cardiopulmonary bypass (CPB) were studied in 54 animals and compared to seven animals without ischemic arrest. Ischemic arrest alone reduced the first derivative of left ventricular force of contraction (LV dF/dt) to 52 percent of control 10 minutes after resuming function and to 64 percent after 1 hour of reperfusion. Cardiac output was depressed to 52 percent of control after 10 minutes of reperfusion, and to 74 percent of control after 60 minutes of reperfusion. In six animals, moderate hypothermia (26 degrees C.) resulted in no protection of cardiac function from ischemic arrest, whereas profound hypothermia to 18 degrees C. resulted in values of LV dF/dt and cardiac output nearly equivalent to the CPB control group (no arrest). A continuous infusion of a hyperkalemic hypothermic solution slightly improved the degree of protection over hypothermia alone. The sarcoplasmic reticulum (SR) isolated from hearts which had undergone 60 minutes of ischemic arrest bound significantly less calcium when the isolation was done after 10 minutes of reperfusion as well as when it was done after 60 minutes of reperfusion. The time to spontaneous release of calcium from the SR also was significantly longer. Moderate hypothermia did not result in improved SR function, whereas deep hypothermia induced by local cooling or by hypothermic potassium infusion retained SR function at normal levels. Oxidative phosphorylation of mitochondria isolated after 60 minutes of reperfusion was also depressed. The mitochondrial respiration rate after normothermic ischemic arrest was 155 natoms of oxygen per minutes versus 237 natoms for the hypothermic hyperkalemic group. Respiratory control index was 5.5 for the normothermic group versus 9.4 for the hypothermic group. It is concluded that hypothermia, whether effected by surface cooling or by hypothermic potassium infusion, allowed full recovery of hemodynamic and biochemical functions within 1 hour of reperfusion.