Degree Of ATP Depletion Research Articles

Myocardial protection with verapamil during ischaemia and reperfusion: dissociation between myocardial salvage and the degree of ATP depletion during ischaemia

The aim was to evaluate the protective effect of verapamil during myocardial ischaemia and reperfusion. In vivo phosphorus-31 (31P) magnetic resonance spectroscopy was performed on rats pretreated with verapamil (mg.kg-1 intraperitoneal) and controls during a 45 min left coronary artery occlusion and 60 min reperfusion. In separate groups of animals, haemodynamic measurements were taken at baseline, during ischaemia, and during reperfusion. Infarct size was determined by staining with triphenyltetrazolium chloride. Female Sprague-Dawley rats were used (control group n = 25, experimental group n = 24). Infarct size was significantly reduced in the verapamil group compared to controls: 9.9(SEM 2.3)%, n = 19 v 28.5(2.7)%, n = 19, p less than 0.001 (infarct % of left ventricular mass). Myocardial phosphocreatine and ATP levels were reduced to similar levels in both verapamil and control animals after 45 min ischaemia: 56.8(3.4)%, n = 10, v 61.4(1.8)%, n = 11 NS; 67.7(2.7)%, n = 10 v 69.7(2.9)%, n = 11, NS (% of baseline value). After 60 min reflow, there was significant recovery of phosphocreatine [91.1(4.2)% of baseline, p less than 0.05] and ATP [86.8(2.7)% of baseline, p less than 0.05] in the verapamil group, but no recovery of high energy phosphates in controls [66.3(2.8), NS; 69.6(2.7), NS]. The left ventricular systolic pressure, heart rate, rate-pressure product, and maximum rate of left ventricular pressure development were similar prior to ischaemia, and during ischaemia in both groups. There was an inverse correlation between infarct size and the degree of phosphocreatine recovery after 60 min of reperfusion (PCr recovery (%) = -0.99 x infarct size (%) + 101; r = 0.91; p less than 0.01; n = 14). Furthermore, in a separate group of animals (n = 9), there was a significant correlation between the size of the ischaemic area at risk and the degree of phosphocreatine decline after 15 min of coronary occlusion (PCr reduction (%) = 0.91 x risk area (%) + 5.6; r = 0.97; p less than 0.01). Pretreatment with verapamil extends the ischaemic time after which reperfusion results in myocardial salvage in this model of ischaemia and reperfusion. This protective effect is independent of the haemodynamic determinants of myocardial oxygen demand and the degree of ATP and phosphocreatine depletion during the ischaemic period. In this model of reversible ischaemia, 31P magnetic resonance spectroscopy is useful for quantitating both the size of the ischaemic region during coronary artery occlusion and infarct size after reperfusion.

Effects of anoxia on K and Ca currents in isolated guinea pig cardiocytes

Whole cell currents were measured in isolated cardiocytes of guinea pig under anoxic conditions (pO2 less than 0.5 torr). After 2 to 32 (mean 11.2) minutes of anoxia, time independent outward currents developed gradually which had a linear current-voltage relation between -100 and +20 mV and reversed at the resting potential of the cells (-82 to -90 mV). After 20 to 170 (mean 38) seconds, the amplitude of these outward currents saturated (3.6 +/- 0.5 nA at +10 mV, n = 23). Reoxygenation within one minute after the appearance of the first extra outward currents led in most cells (greater than 90%) to their complete disappearance in 2 to 4 (mean 2.87, n = 15) seconds. Ca currents were not affected at the time when the first extra outward currents occurred. It is concluded that (i) the anoxia-induced outward current is carried by K+ ions probably through KATP channels which open at intracellular ATP concentrations below 1 mmol/l (Noma and Shibasaki 1985) and (ii) this degree of ATP depletion does not affect normal Ca channel function.

Intracellular ATP in a glucosephosphate isomerase mutant of Saccharomyces cerevisiae.

The degree of ATP depletion caused by glucose in a glucosephosphate isomerase-deficient strain of Saccharomyces cerevisiae was determined. Even in the presence of a sugar normally fermentable by the mutant, the addition of glucose can decrease the intracellular ATP, depending on the competition of the sugars for transport and subsequent phosphorylation. For both parent and mutant cells, a correlation exists between the calculated velocity of ATP formation or ATP consumption during the utilization of different concentrations of sugars and the experimental intracellular ATP level. For initially resting yeast cells, a rate increase of 35 mumol per min per g ATP was calculated to increase the intracellular level of this nucleotide by 1 mumol per g cell mass.

Disparate mechanisms for hypoxic cell injury in different nephron segments. Studies in the isolated perfused rat kidney.

Hypoxic injury was evaluated morphologically in the proximal tubule and in the medullary thick ascending limb of isolated rat kidneys perfused for 90 min without O2 or with various metabolic inhibitors. Inhibition of mitochondrial respiration (with rotenone, antimycin, oligomycin) or of intermediary metabolism (with monofluoroacetate, malonate, 2-deoxyglucose) caused reduction in renal oxygen consumption, renal function, and ATP content comparable with those elicited by oxygen deprivation. Metabolic inhibition produced hypoxiclike injury in the first portions of the proximal tubule, S1 and S2 ("clubbing" of microvilli, mitochondrial swelling), and the extent of damage was correlated with the degree of ATP depletion. In the third portion of the proximal tubule, S3, hypoxiclike damage (cytoplasmic edema or fragmentation) occurred most consistently when both aerobic and anaerobic metabolism were inhibited simultaneously. In the medullary thick ascending limb, none of the metabolic or mitochondrial inhibitors used could reproduce the injury of oxygen deprivation. Thus, the proximal tubule and the thick ascending limb have markedly different responses to cellular energy depletion, suggesting disparate mechanisms for hypoxic injury along the nephron.

Neurologic deficit and cerebral ATP depletion after temporary focal ischemia in cats.

Focal ischemia was induced in 23 cats by occluding the left middle cerebral artery for 2 h. The animals were then divided into groups for unforced reperfusion of variable duration ranging from 2 to 48 h. Neurological ratings were obtained during both ischemia and reperfusion. Following planned sacrifice the regional ATP content was assessed by means of a bioluminescence method showing spatial distribution and degree of ATP depletion. All the animals developed a neurologic deficit, with a median of 6 points on a disability scale of 0-10. After reopening of the middle cerebral artery, neurologic recovery was quite variable depending on the initial neurologic deficit (partial phi = 0.67, p1 less than 0.05): Animals with mild initial functional impairment improved and those with severe neurologic disturbances either died early or developed a more severe neurologic deficit, irrespective of the duration of reperfusion. The degree of ATP depletion and the amount of brain tissue involved exhibited a significant correlation with the neurological outcome (tau = 0.50, p1 less than 0.05), but they were even more closely related to the initial neurologic deficit (partial phi = 1.00, p1 less than 0.001), suggesting an early definitive manifestation of deficiencies in regional energy metabolism.