1 The cardioprotective properties of inhibition of poly (ADP-ribose) synthetase (PARS) were investigated in the isolated perfused heart of the rat. Hearts were perfused in the Langendor mode and subjected to 23 min total global ischaemia and reperfused for 60 min. 2 Left ventricular function was assessed by means of an intra-ventricular balloon. High energy phosphates were measured by 31 P-NMR spectroscopy. Intracellular levels of NAD were measured by capillary electrophoresis of perchloric acid extracts of hearts at the end of reperfusion. 3 Reperfusion in the presence of the PARS inhibitor 1,5 didroxyisoquinoline (ISO, 100 mM) attenuated the mechanical dysfunction observed following 1 h of reperfusion; 27+13 and 65+8% recovery of preischaemic rate pressure product for control and 100 mM ISO, respectively. 4 This cardioprotection was accompanied by a preservation of intracellular high-energy phosphates during reperfusion; 38+2 vs 58+4% (P50.05) of preischaemic levels of phosphocreatine (PCr) for control and 100 mM ISO respectively and 23+1 vs 31+3% (P50.05) of preischaemic levels of ATP for control and 100 mM ISO respectively. 5 Cellular levels of NAD were higher in ISO treated hearts at the end of reperfusion; 2.56+0.45 vs 4.76+1.12 mmoles g 71 dry weight (P50.05) for control and ISO treated. 6 These results demonstrate that the cardioprotection a orded by inhibition of PARS activity with ISO is accompanied by a preservation of high-energy phosphates and cellular NAD levels and suggest that the mechanism responsible for this cardioprotection may involve prevention of intracellular ATP depletion.

The aim of this work was to investigate whether 1) Rb uptake is reduced in ischemic myocardium, and 2) 87Rb three-dimensional (3D) imaging can detect the ischemic area. Hearts of domestic pigs (n = 8, 20-30 kg) were perfused retrogradely with Krebs-Henseleit buffer in a 7-T, 40-cm horizontal bore magnet interfaced with Bruker MSLX spectrometer. Control (C) and ischemic (I) (45 min ligation of the left anterior descending coronary artery (LAD)) hearts were loaded with Rb+ by perfusion with a Rb(+)-containing solution (2-4.7 mM, 30-100% of K+ substitution) for 35 min and 87Rb (C) or 31P (ischemic area in I) spectra were acquired. After mechanical arrest with 0.6-0.9 mM lidocaine, 87Rb images (7 min each, 1 cm3 resolution) were acquired (30-40 min) in the presence of Rb(+)-containing perfusate. Subsequently, the hearts were stained with Evans blue (EB) and samples taken for measurements of Rb+ content. In the Group C, distribution of Rb+ in the left ventricle and the intensities of the 3D 87Rb images were uniform. In the ischemic area (Group I), verified by the lack of EB staining and changes in 31P spectra, the images showed a reduced intensity, which corresponded to decreased Rb+ content (33 +/- 11% of the normal). Thus, 87Rb imaging reveals damaged cells detecting reduced Rb+ content in the ischemic area.

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87Rb-MRI was used to measure Rb(+) uptake in blood-perfused pig hearts during complete occlusion (120 and 70 min) of the left anterior descending artery (LAD) and subsequent reperfusion (120 and 170 min). The Rb(+) uptake rate and maximal Rb image intensity during 120-min occlusion were significantly lower in the ischemic anterior left ventricular (LV) wall (0.35 +/- 0.14%/min and 26 +/- 4.7%) relative to those in the remote posterior LV wall (2.43 +/- 0.33%/min and 98 +/- 10%). Reperfusion after 120 and 70 min of occlusion resulted in formation of damaged areas, which had 40 +/- 4 and 73 +/- 10% of the (87)Rb image intensity observed in the remote posterior wall. The infarct sizes determined histologically by triphenyltetrazolium chloride staining were 9.6 +/- 3.4 and 5.6 +/- 4.6% of the total ventricular mass (LV + RV) in the 120- and 70-min occlusion groups, respectively. The sizes determined by MRI were 13.1 +/- 2 and 2.8 +/- 4.3% of the total number of pixels, respectively. The Rb(+) uptake in the anterior wall during 120-min occlusion was somewhat lower than that previously observed in crystalloid-perfused hearts. It is concluded that blood does not interfere with the ability of (87)Rb MRI to detect ischemic and infarct areas.

Fluorescence, absorbance, and binding of a mitochondrial membrane potential-sensitive probe, rhodamine 800 (rhod800), were measured in isolated rat mitochondria, hepatocytes, cardiomyocytes, and hearts in the presence or absence of mitochondrial uncouplers. Excitation of rhod800 was achieved with laser diodes (690 or 670 nm) and resulted in a fluorescence peak at 720 nm. Greater than 99% of rhod800 (1 microM) was taken up from the buffer by energized mitochondria. This resulted in a fluorescence decrease by 77% (13% in de-energized mitochondria). Sixty-seven percent of rhod800 was taken up by cardiomyocytes and 75% by hepatocytes resulting in the fluorescence decrease by 16% and 37%, respectively, which were reversed by approximately 10% upon cell uncoupling. In hearts, binding, absorbance, and fluorescence were almost uncoupler-insensitive possibly due to rhod800 interaction outside of mitochondria. Fluorescence of the hearts perfused with 27.5 and 55 nM rhod800 was measured in orthogonal and reflection modes. The former provided deep tissue penetration (approximately a centimeter); however, nonlinearity between absorbance and fluorescence was evident. In the latter setting, depth of tissue penetration was approximately a millimeter, which eliminated an inner filter effect and restored linearity. We concluded that excessive hydrophobicity of rhod800 complicates detection of energy-dependent fluorescence changes in myocardium.

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