Preservation injury and acute rejection of rat intestinal grafts: protection afforded by pyruvate.

Pyruvate has been shown to prevent intestinal mucosal injury after ischemia-reperfusion. The aim of the present study was to determine whether pyruvate can (1) prevent postreperfusion mucosal injury occurring after intestinal preservation and subsequent transplantation and (2) exert a protective effect on the intestinal graft mucosa during acute rejection. Preservation mucosal injury was evaluated, after 2 hours of reperfusion, by comparing grafts transplanted in a rat syngeneic combination (ACI to ACI) after 2 hours of cold preservation using pyruvate (n = 6) or placebo (n = 6). Mucosal parameters obtained during acute rejection (allogeneic combination: ACI to Lewis) were compared between placebo-treated (n = 6) and pyruvate-treated (n &equals 6) animals. Tissue injury was evaluated by histopathologic examination, oxygen free radical production by luminol-enhanced chemiluminescence, and degree of neutrophil infiltration by myeloperoxidase staining. After reperfusion of the preserved grafts and during acute rejection, mucosal oxygen free radical levels and the number of infiltrating neutrophils were significantly (P <0.05) increased in the untreated grafts, whereas there was a statistically significant inhibition of these parameters in those treated with pyruvate. Mucosal injury, seen after reperfusion of the preserved grafts, was prevented by pyruvate. The histopathologic abnormalities observed in the untreated grafts during rejection were also significantly reduced by pyruvate. Treatment with pyruvate before cold preservation of intestinal grafts, in this rat model, reduced reperfusion mucosal injury, neutrophil infiltration, and oxygen free radical production. Oxygen free radicals were produced in the mucosa of the graft during acute rejection and their production was reduced by pyruvate, which exerted a protective effect on the rejecting allograft mucosa.

Inhibition of regain in body weight and fat with addition of 3-carbon compounds to the diet with hyperenergetic refeeding after weight reduction.

OBJECTIVE: To investigate the efficacy of the 3-carbon compounds pyruvate and dihydroxyacetone (PD) in inhibiting reaccumulation of body weight and fat with refeeding after weight loss. DESIGN: Longitudinal, in Clinical Research Center. After weight loss induced by hypoenergetic diet (1.3 MJ/d) for 3 weeks, refeeding with hyperenergetic diet (1.5 x resting energy expenditure) for 3 weeks. Refeeding diet randomized to contain PD or placebo (PL, polyglucose) as approximately 20% of energy intake. SUBJECTS: 17 obese healthy women (n = 8 in PL group, n = 9 in PD group) (age: 22-60 y, weight: 72.5-139.7 kg). MEASUREMENTS: Resting energy expenditure (REE), body composition (by bioelectrical impedance), nitrogen balance, serum proteins, biochemical profile, thyroid hormones, and insulin, before and after refeeding and weight and fat gain. RESULTS: Refeeding with a hyperenergetic diet, weight gain was significantly less in patients receiving PD compared to placebo (1.8 + 0.2 kg vs 2.9 +/- 0.1 kg, P < 0.01). Body fat regain was also less with feeding of PD (0.8 +/- 0.2 kg vs 1.8 +/- 0.2 kg, P < 0.01). Body protein metabolism, as measured by nitrogen balance, serum protein concentrations and fat free mass, was similar in subjects consuming either PD or PL. CONCLUSIONS: We conclude that 3-carbon compounds decrease weight gain and reaccumulation of body fat, without decreasing body protein gain, in obese subjects with hyperenergetic refeeding subsequent to weight loss.

Pyruvate reduces anoxic injury and free radical formation in perfused rat hepatocytes.

The effects of 5 mM pyruvate on anoxic injury, superoxide (O2-.) and hydrogen peroxide (H2O2) generation, and lactate dehydrogenase (LDH) release during reoxygenation after 2.5 h anoxia were studied in perfused rat hepatocytes. When pyruvate was present during anoxia and reoxygenation, there was little anoxic injury, and the generation of free radicals and LDH release during reoxygenation were reduced 50-60%. When Pyruvate was added during reoxygenation, there was no decrease in O2-. or LDH release, although H2O2 formation was depressed. Free radical formation and anoxic/reperfusion injury were significantly reduced when pyruvate was added during the anoxic period only. Pyruvate reduced the deleterious effects of 10 microM antimycin A by preventing the increase in O2-. formation and LDH release evoked by the inhibitor. These results indicate that pyruvate protected hepatocytes against anoxic injury and that its protective action occurred principally during anoxia and not during reoxygenation. Pyruvate appeared to act at a mitochondrial site, since it reduced the deleterious effects of antimycin A.

Hemodynamic effects of pyruvate infusion in dogs.

PURPOSE: Although pyruvate supplementation enhances endurance in humans and increases cardiac output in dogs, its effects on cardiac and peripheral vascular function are not known. Thus, we assessed the cardiovascular effects of pyruvate infusion. MATERIALS AND METHODS: Aortic, left ventricular (LV), and pulmonary (Ppa) pressures and LV stroke volume (Svlv; derived from aortic flow probe) were measured after thoracotomy in eight anesthetized dogs. LV area or volume changes were measured using either an epicardial echocardiography (n = 6) or a conductance catheter (n = 2). LV end-systolic elastance (Eeslv) and preload recruitable stroke force (PRSFlv) relations, as estimates of contractility, were generated by transient inferior vena cava occlusion. Simultaneous stroke volume to arterial pressure relations during the occlusions were used to measure arterial elastance (Ea), and steady-state systemic and pulmonary vascular resistances were used as measures of arterial tone. Graded doses of pyruvate (8, 16, and 32 mg/kg/min), dobutamine (positive control) and propranolol (negative control) and placebo (volume control) were sequentially given. RESULTS: Dobutamine increased Eeslv, PRSFlv, whereas propranolol had the opposite effect on Eeslv and PRSFlv. Pyruvate at 32 mg/kg/min increased heart rate, Ppa, and SVlv and decreased LV end-diastolic area, and systemic vascular resistance without changing arterial pressure, Eeslv, PRSFlv, or Ea. CONCLUSIONS: We conclude that pyruvate infusion in normal dogs induces venodilation but does not alter either cardiac contractility or arterial tone.

Pyruvate inhibits clofibrate-induced hepatic peroxisomal proliferation and free radical production in rats.

In an effort to identify the effects of the 3-carbon compound pyruvate on free radical production, we measured hepatic total peroxisomal beta-oxidation and catalase activity and the production of lipofuscin-like products in male Sprague-Dawley rats consuming an adequate diet supplemented with pyruvate, vitamin E, or the peroxisome proliferator and free radical enhancer clofibrate for 22 days (n = 5 in each group). Clofibrate feeding induced hepatomegaly, a fivefold increase in total peroxisomal beta-oxidation activity, and a threefold increase in hepatic lipofuscin-like products (P < .05). Pyruvate but not vitamin E inhibited the increase in liver size by 70% (P < .05). Both pyruvate and vitamin E completely inhibited clofibrate-induced increases in lipofuscin-like products (P < .05). Pyruvate but not clofibrate or vitamin E increased plasma concentrations of the nitric oxide metabolites nitrite and nitrate (P < .05). We conclude that with clofibrate-induced peroxisomal proliferation and free radical production, pyruvate will inhibit peroxisomal proliferation and free radical production, inhibit free radical-induced lipid peroxidation, and enhance metabolism of nitric oxide.

Hemodynamic effects of intravenous pyruvate in the intact, anesthetized dog.

OBJECTIVES: There is evidence from human studies that pyruvate improves skeletal muscle endurance, and from isolated heart preparations that pyruvate is a positive inotrope. We examined the hemodynamic effects of intravenous pyruvate in an intact, anesthetized dog preparation in order to test its effects in an intact animal. Our hypothesis was that pyruvate is a positive inotrope in the intact dog. DESIGN: Prospective, randomized, controlled trial. SETTING: Animal laboratory. SUBJECTS: Ten mongrel dogs. INTERVENTIONS: Two groups of animals were anesthetized with chloralose and urethane, mechanically ventilated, and hemodynamically monitored. The experimental group (n = 6) received an infusion of calcium pyruvate and sodium pyruvate, while the control group (n = 4) received an infusion of calcium chloride and sodium chloride. MEASUREMENTS AND MAIN RESULTS: The intravenous infusion of calcium and sodium pyruvate resulted in increased cardiac output, left ventricular contractility, and mixed venous oxygen saturation values in the experimental group compared with the control group of four dogs. There were no significant detrimental effects except an increase in the mean serum calcium concentrations in both groups. CONCLUSIONS: These data suggest that intravenous pyruvate may be a useful in vivo positive inotrope.

Pyruvate inhibits growth of mammary adenocarcinoma 13762 in rats.

The growth of implanted mammary adenocarcinoma 13762 was measured in rats consuming a liquid diet (35% fat, 18% protein, 47% carbohydrate) supplemented with pyruvate (37.3 g/liter; n = 13) or maltose-dextrin (placebo; n = 13) for 21 days. Mean tumor diameter, measured on day 11, 14, 18, and 21 subsequent to tumor implantation, was 41, 32, 21, and 19% smaller in the pyruvate group (P < 0.05). When euthanized, tumor weight was also smaller in the pyruvate group: pyruvate = 15.0 +/- 2.3 (SEM) g; placebo = 24.9 +/- 3.2 g, P < 0.05. Visual inspection of organs suggested decreased lung metastases with pyruvate feeding (P < 0.05). Upon microscopic evaluation of organs, hepatic tumor was found only in the placebo group. We conclude that pyruvate inhibits implanted tumor growth in rats.

Pyruvate supplementation of a low-cholesterol, low-fat diet: effects on plasma lipid concentrations and body composition in hyperlipidemic patients.

The effects of the three-carbon compound pyruvate on plasma lipid concentrations and body composition were evaluated in hyperlipidemic patients consuming a low-cholesterol (165-180 mg), low-fat (22-24% of energy; 18-20% of energy as saturated fatty acid) diet (0.091-0.099 MJ.kg body wt-1 x d-1). After consuming the above diet for 4 wk, during which time plasma lipid concentrations decreased, 34 subjects were randomly assigned to receive either 22-44 g pyruvate (n = 17) or 18-35 g polyglucose (placebo, Polycose, n = 17), iso-energetically substituted for a portion of carbohydrate energy for 6 wk. Despite greater weight and fat losses with pyruvate (P < 0.05), plasma concentrations of cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride were not different between the two groups of subjects. We conclude that subsequent to diet-induced reduction in plasma lipid concentrations, pyruvate supplementation of a low-cholesterol, low-fat diet providing 6.7-7.6 MJ/d for 6 wk has no effect on plasma lipid concentrations but enhances body weight and fat losses.

Amino acid arterial concentration and muscle exchange during submaximal arm and leg exercise: the effect of dihydroxyacetone and pyruvate.

The mixture of dihydroxyacetone and pyruvate (DHAP) is an ergogenic aid that enhances muscle glucose extraction during prolonged aerobic exercise. In order to evaluate the effect of DHAP on muscle amino acid extraction during exercise, we measured arterial concentration and muscle exchange of amino acids in 18 untrained healthy male subjects (aged 20-30 years) performing dynamic arm (60% VO2 max, n = 9) or leg (70% VO2 max, n = 9) exercise to exhaustion with and without dietary supplementation of DHAP. The subjects consumed diets (146 kJ kg body weight-1 day-1) containing either 100 g polyglucose, Polycose (placebo, P) or DHAP (3:1, treatment) substituted for a portion of carbohydrate. The two diets were administered in a double-blind, random, crossover order for a 7-day period. At least 7 days separated the dietary protocols. Blood samples were drawn through radial artery and axillary or femoral vein catheters at rest, during exercise and at exhaustion. Arterial alanine concentration increased by 30% during arm exercise and by 50-60% during leg exercise. No other arterial amino acid concentration changed during exercise. At exhaustion, arterial alanine concentration decreased to pre-exercise levels with arm exercise but remained elevated after leg exercise. Despite changes in arterial concentrations of alanine with exercise, muscle exchange of alanine was not altered with exercise. Exercise did not alter muscle exchange of any amino acid. Arterial amino acid concentrations and muscle exchange of amino acids with exercise were similar with or without DHAP feeding.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma lipid concentrations in hyperlipidemic patients consuming a high-fat diet supplemented with pyruvate for 6 wk.

We evaluated the effects of a three-carbon compound, pyruvate, on plasma lipid concentrations in hyperlipidemic patients consuming a high-cholesterol (560-620 mg), high-fat (45-47% of energy; 18-20% of energy as saturated fatty acid), anabolic diet (0.11-0.12 MJ/kg body wt) for 6 wk. Forty subjects consumed the diet, randomly supplemented with 36-53 g pyruvate (n = 19) or 21-37 g polyglucose (placebo, Polycose, n = 21) as a portion of carbohydrate energy. Plasma cholesterol and LDL-cholesterol concentrations were unchanged in the placebo group, but decreased by 4% and 5%, respectively, in the pyruvate group (P < 0.05 vs placebo). Plasma HDL-cholesterol, HDL3-cholesterol, and triglyceride concentrations were similar in both groups. Resting heart rate, diastolic blood pressure, and rate-pressure product were unchanged after 6 wk of therapy in the placebo group, but decreased by 9%, 6%, and 12%, respectively with pyruvate supplementation (P < 0.05 vs placebo). We conclude that pyruvate supplementation of a high-fat, high-cholesterol, anabolic diet will decrease plasma cholesterol and LDL-cholesterol concentrations without affecting the HDL-cholesterol concentration.

Body composition, energy utilization, and nitrogen metabolism with a 4.25-MJ/d low-energy diet supplemented with pyruvate.

We measured body composition, energy deficit, and nitrogen metabolism in 14 obese women housed in a metabolic ward, who consumed a 4.25-MJ/d liquid diet (68% carbohydrate, 22% protein) for 21 d with or without pyruvate (PY; n = 7) partially, isoenergetically substituted for glucose (placebo; n = 7). Body composition and leucine oxidation and turnover were determined before and after weight loss. Energy deficit was calculated from resting metabolic rates. Subjects fed pyruvate showed a greater weight loss (PY = 5.9 +/- 0.7 kg, placebo = 4.3 +/- 0.3 kg, P less than 0.05), fat loss (PY = 4.0 +/- 0.5 kg, placebo = 2.7 +/- 0.2 kg, P less than 0.05), kg wt loss/4.25-MJ deficit (PY = 0.22 +/- 0.01 kg, placebo = 0.17 +/- 0.01 kg, P less than 0.05, and kg fat loss/4.25-MJ deficit (PY = 0.15 +/- 0.01 kg, placebo = 0.11 +/- 0.01 kg, P less than 0.05). Nitrogen balance (urine and stool) and leucine oxidation and turnover were similar in both groups. We conclude that the dietary modification whereby the three-carbon compound pyruvate is isoenergetically substituted for the six-carbon compound glucose in a 4.25-MJ/d, low-energy diet will increase fat and weight loss.

Body composition, nitrogen metabolism, and energy utilization with feeding of mildly restricted (4.2 MJ/d) and severely restricted (2.1 MJ/d) isonitrogenous diets.

To determine the effects on weight loss of feeding isonitrogenous diets in mildly restricted (4.2 MJ/d) and severely restricted (2.1 MJ/d) amounts, we measured body composition, weight loss-energy deficit ratio, and nitrogen metabolism in 14 obese women housed in a metabolic ward consuming hypoenergetic diets for 21 d. Subjects consumed either a 4.2-MJ/d diet (50 g protein, 175 g carbohydrate) or a 2.1-MJ/d diet (50 g protein, 75 g carbohydrate). Body composition and leucine oxidation and turnover were determined before and after weight loss. Energy deficit was calculated from resting metabolic rates. Subjects fed the 2.1-MJ/d diet showed a greater weight loss (6.1 +/- 0.5 vs 4.5 +/- 0.5 kg; mean +/- SE, P less than 0.05) and fat loss (3.9 +/- 0.3 vs 3.0 +/- 0.3 kg, P less than 0.05). Weight loss-energy deficit ratio was the same with both diets. Nitrogen balance and leucine oxidation and turnover were similar in both groups. We conclude that with feeding of isonitrogenous hypoenergetic diets, severe restriction of energy content (2.1 MJ/d, 75 g carbohydrate) will enhance weight and fat loss without increasing nitrogen loss compared with mild restriction of energy (4.2 MJ/d).

Body composition, energy utilization, and nitrogen metabolism with a severely restricted diet supplemented with dihydroxyacetone and pyruvate.

To determine the effect of dietary modification on energy utilization during severely restrictive hypocaloric feeding, we measured body composition, energy deficit, and nitrogen metabolism in 13 obese women housed in a metabolic ward consuming a 2.1-MJ diet for 21 d with the three-carbon compounds dihydroxyacetone and pyruvate (DHAP), partially, isocalorically substituted for glucose. Body composition and amino acid (leucine) oxidation and turnover were determined before and after weight loss. Energy deficit was calculated from metabolic rates and compared with weight and fat loss. Subjects fed dihydroxyacetone and pyruvate showed a greater weight loss (DHAP = 6.5 +/- 0.3 kg, P = 5.6 +/- 0.2 kg), fat loss (DHAP = 4.3 +/- 0.2 kg, P = 3.5 +/- 0.1 kg), and weight and fat loss/4.25-MJ deficit (P less than 0.05 for all determinations). Nitrogen balance (urine and stool) and leucine metabolism were similar in both groups. We conclude that partial substitution of DHAP for six-carbon compounds of a 2.1-MJ diet will increase weight and fat loss.

Enhanced leg exercise endurance with a high-carbohydrate diet and dihydroxyacetone and pyruvate.

The effects of dietary supplementation of dihydroxyacetone and pyruvate (DHAP) on metabolic responses and endurance capacity during leg exercise were determined in eight untrained males (20-30 yr). During the 7 days before exercise, a high-carbohydrate diet was consumed (70% carbohydrate, 18% protein, 12% fat; 35 kcal/kg body weight). One hundred grams of either Polycose (placebo) or dihydroxyacetone and pyruvate (treatment, 3:1) were substituted for a portion of carbohydrate. Dietary conditions were randomized, and subjects consumed each diet separated by 7-14 days. After each diet, cycle ergometer exercise (70% of peak oxygen consumption) was performed to exhaustion. Biopsy of the vastus lateralis muscle was obtained before and after exercise. Blood samples were drawn through radial artery and femoral vein catheters at rest, after 30 min of exercise, and at exercise termination. Leg endurance was 66 +/- 4 and 79 +/- 2 min after placebo and DHAP, respectively (P less than 0.01). Muscle glycogen at rest and exhaustion did not differ between diets. Whole leg arteriovenous glucose difference was greater (P less than 0.05) for DHAP than for placebo at rest (0.36 +/- 0.05 vs. 0.19 +/- 0.07 mM) and after 30 min of exercise (1.06 +/- 0.14 vs. 0.65 +/- 0.10 mM) but did not differ at exhaustion. Plasma free fatty acids, glycerol, and beta-hydroxybutyrate were similar during rest and exercise for both diets. Estimated total glucose oxidation during exercise was 165 +/- 17 and 203 +/- 15 g after placebo and DHAP, respectively (P less than 0.05). It is concluded that feeding of DHAP for 7 days in conjunction with a high carbohydrate diet enhances leg exercise endurance capacity by increasing glucose extraction by muscle.