Inhibition of hydroxyl radical production by lactobionate, adenine, and tempol.

Superoxide and hydroxyl free radicals are strongly implicated in the deleterious effects of reperfusion of grafted organs. Iron ions are critical in the Fenton-like reaction that generates oxygen-free radicals from H2O2. Using the ADP/Fe2+/H2O2 .OH-generating system, we demonstrated that components of an organ-preservation solution (Henri Mondor solution): sodium lactobionate, adenine, and a nitroxide radical: 4-hydroxy-2,2,6,6-tetramethylpiperidine-n-oxyl (TEMPOL), showed unexpected inhibition properties on the production of hydroxyl radicals by complexation of Fe2+ for lactobionate and nitroxide or Fe3+ for adenine. This inhibition was 75.5% at 12 mM lactobionate. Moreover, a complete inhibition was observed at 50 mM. At 0.25 mM adenine, the reduction was 14.8% (maximum effect: 34.1%). Henri Mondor solution, at an identical adenine and lactobionate concentration, inhibited the radical production by 91.5%, indicating an additive effect. Nitroxide totally inhibited .OH production by the ADP/Fe2+/H2O2 system (maximum effect: 95.6%) and partially the production by an O2.- generating system (maximum effect: 74.8%). Thus, the association of these three components in preservation solutions would be an original method to limit the reperfusion injury observed in isolated ischemic organs.

In vivo comparative study of two lactobionate based solutions for prolonged heart preservation.

The duration of safe heart preservation must be improved. Using a heterotopic heart transplantation model, we compared in vivo the recovery of rabbits hearts preserved with a K+Lactobionate based fluid (UW: University of Wisconsin solution) or with a Na+Lactobionate based fluid. In the "preservation" group, hearts were cold stored (4 degrees C) for 6 hours with UW (n = 9) or Na+Lactobionate solution (n = 9). In the "transplantation" group, cold storage was followed by 3 hours of reperfusion (UW: n = 8, Na+Lactobionate solution: n = 7). Functional recovery, adenine nucleotide pool, circulating blood cardiac enzymes, circulating blood and tissue malondialdehyde (MDA) were measured. Left ventricular end-diastolic and developed pressures at different preload levels were better after preservation with UW than with Na+Lactobionate solution (p < 0.05). Also with UW, adenosine diphosphate and total adenine nucleotide content were significantly higher than with Na+Lactobionate solution (p < 0.05) whereas adenosine triphosphate, monophosphate and energy charges were similar. Cardiac enzymes and tissue MDA were similar with UW and Na+Lactobionate solution. In circulating blood, MDA was not detected. These results enhance the superiority of UW solution over a Na+Lactobionate based solution for long term heart preservation.

Myocardial high-energy phosphate depletion in allograft rejection after orthotopic human heart transplantation.

The present study was designed to assess whether acute rejection affects myocardial energy content of the human orthotopically transplanted heart. Adenosine triphosphate content was measured in one tissue sample obtained during 46 routine right ventricular endomyocardial biopsies 6 to 455 days (98 +/- 110) after transplantation in 19 cyclosporine-treated transplant recipients. Tissue samples were immediately frozen in liquid nitrogen within 10 seconds after excision. Adenosine triphosphate analysis was performed with high performance liquid chromatography. Three groups of biopsy specimens were classified according to the standardized cardiac biopsy grading system. Group 1: Eight biopsy specimens without rejection; group 2: 24 biopsy specimens with mild rejection; group 3: 14 biopsy specimens with moderate or severe rejection. Graft systolic function evaluated by echocardiographic fractional shortening was in the normal range the day of biopsy. All patients had normal coronary angiograms within 1 month of the study. In the presence of mild rejection (grade 1A or 1B), adenosine triphosphate content was not significantly different from that of nonrejecting hearts (26.15 +/- 7.1 and 28.57 +/- 8.23 nmol/mg protein, respectively). By contrast, a significant decrease in adenosine triphosphate content was observed when moderate or severe rejection with focal or diffuse aggressive infiltrates were present (10.46 +/- 4.11 nmol/mg protein; p < 0.01 versus two other groups). In seven cases, sequential analysis showed a significant increase in adenosine triphosphate content after rejection therapy concomittant with histologic improvement: 10.19 +/- 2.9 before and 30.13 +/- 7.0 nmol/mg protein after treatment (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Reduced production of the deleterious hydroxyl free radical during the final reperfusion of isolated rabbit heart with the use of an improved sodium lactobionate-based cardioplegic medium.

The production of free radicals during the reperfusion of hearts after prolonged ischemia might be responsible for the deleterious effects observed. Several strategies have attempted to limit this production or trap the radicals produced. This study aimed to evaluate the efficacy of a sodium lactobionate-based solution supplemented with 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl, a stable nitroxide free radical, in scavenging hydroxyl radicals produced during the reperfusion of the isolated rabbit heart. 4-Hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl was administered just before the end of the storage period (6 hours) in a cardioplegic solution, either crystalloid solution or sodium lactobionate-based solution. The production of hydroxyl radicals was assessed through the production of dihydroxybenzoic acid, a reaction compound of hydroxyl radicals with salicylate. Two main isomers of dihydroxybenzoic acid were found in the coronary effluents: 2,3 and 2,5 dihydroxybenzoic acid. The production was 40-fold lower when the hearts received sodium lactobionate-based solution than when they received crystalloid solution (p < 0.001). The presence of 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl had no influence on the production of hydroxyl radicals. The production of hydroxyl radicals was maximal during the first minute of the reperfusion. Iron ions, directly involved in the production of hydroxyl radicals, were shown to be complexed by sodium lactobionate in vitro. These results underlie the important role of lactobionate in the prevention of the reperfusion injury and the inefficacy of 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl to decrease the production of hydroxyl radicals.