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We have recently demonstrated that endogenous H2O2 plays an important role in coronary autoregulation in vivo. However, the role of H2O2 during coronary ischemia-reperfusion (I/R) injury remains to be examined. In this study, we examined whether endogenous H2O2 also plays a protective role in coronary I/R injury in dogs in vivo. Canine subepicardial small coronary arteries (≥100 μm) and arterioles (G-monomethyl-l-arginine (l-NMMA), catalase (a decomposer of H2O2), 8-sulfophenyltheophylline (8-SPT, an adenosine receptor blocker), l-NMMA + catalase, l-NMMA + tetraethylammonium (TEA, an inhibitor of large-conductance Ca2+-sensitive potassium channels), and l-NMMA + catalase + 8-SPT. Coronary I/R significantly impaired the coronary vasodilatation to ACh in both sized arteries (both P < 0.01); l-NMMA reduced the small arterial vasodilatation (both P < 0.01), whereas it increased ( P < 0.05) the ACh-induced coronary arteriolar vasodilatation associated with fluorescent H2O2 production after I/R. Catalase increased the small arterial vasodilatation ( P < 0.01) associated with fluorescent NO production and increased endothelial NOS expression, whereas it decreased the arteriolar response after I/R ( P < 0.01). l-NMMA + catalase, l-NMMA + TEA, or l-NMMA + catalase + 8-SPT further decreased the coronary vasodilatation in both sized arteries (both, P < 0.01). l-NMMA + catalase, l-NMMA + TEA, and l-NMMA + catalase + 8-SPT significantly increased myocardial infarct area compared with the other four groups (control, l-NMMA, catalase, and 8-SPT; all, P < 0.01). These results indicate that endogenous H2O2, in cooperation with NO, plays an important cardioprotective role in coronary I/R injury in vivo.

Exogenous nitric oxide (NO) suppresses endothelium-derived NO production. We were interested in determining whether this is also the case in flow-induced endothelium-derived NO production. If so, then is the mechanism because of intracellular depletion of tetrahydrobiopterin [BH4; a cofactor of NO synthase (NOS)], which results in superoxide production by uncoupled NOS? Isolated canine femoral arteries were perfused with 100 μM S-nitroso- N-acetylpenicillamine (SNAP; an NO donor) and/or 64 μM BH4. Perfusion of SNAP suppressed flow-induced NO production, which was evaluated as a change in the slope of the linear relationship between perfusion rate and NO production rate ( P < 0.02 vs. control; n = 7). Subsequent BH4perfusion returned the slope to the control level. Concomitant perfusion of SNAP and BH4retained the control-level NO production ( n = 7). Concomitant perfusion of SNAP and 4,5-dihydroxy-1,3-benzene disulfonic acid (Tiron; 1 mM; a membrane-permeable superoxide scavenger) also retained the control-level NO production ( n = 7), whereas perfusion of Tiron after SNAP could not return the NO production to the control level ( P < 0.02 vs. control; n = 7). We also found a significant decrease in BH4concentration in the endothelial cells after SNAP perfusion. In conclusion, these results indicate that exogenous NO suppresses the flow-induced, endothelium-derived NO production by superoxide released from uncoupled NOS because of intracellular BH4depletion.

Time-sequential responses to endothelium-dependent and -independent vasodilators and angiotensin-converting enzyme (ACE) inhibitors were studied in the subendocardial arterioles (Endo) of canine renovascular hypertension (HT) compared with subepicardial arterioles (Epi; both

The presence of a coronary stenosis results primarily in subendocardial ischemia. Apart from the decrease in coronary perfusion pressure, a stenosis also decreases coronary flow pulsations. Applying a coronary perfusion system, we compared the autoregulatory response of subendocardial ( n = 10) and subepicardial ( n = 12) arterioles (G-monomethyl-l-arginine abrogated the effect of the stenosis on flow. We conclude that the decrease in pressure caused by a stenosis in vivo results in a larger decrease in diameter of the subendocardial arterioles than in the subepicardial arterioles, and furthermore stenosis selectively decreases the dilatory response of subendocardial arterioles. These two findings expand our understanding of subendocardial vulnerability to ischemia.

The goal of this study was to evaluate microheterogeneity of myocardial blood flow and its dependence on arterial O2 tension (PaO2). We measured within-layer distribution of regional blood flows in the left ventricles of anesthetized rabbits in both normoxic and hypoxic states with myocardial region sizes in the range of 0.01-1.0 mm2. A novel method of digital radiography combined with the technique of 3H-labeled desmethylimipramine deposition enabled us to visualize and accurately quantitate regional blood flow at such high levels of resolution. To analyze myocardial blood flow patterns, we computed the coefficient of variation (CV) and the correlation between adjacent regional flows (CA). The CA values were larger in the hypoxic state (PaO2 = 26 +/- 5 mmHg) than in the normoxic state (PaO2 = 97 +/- 20 mmHg) at all levels of resolution (P < 0.001). In the normoxic state, there was a transmural difference in CA (P < 0.001); CA increased with depth of the left ventricle (from subepicardium to subendocardium). However, the relation between CA and the depth of the left ventricle was not statistically significant in the hypoxic state. The CV values were smaller in the hypoxic state than in the normoxic state at all levels of resolution (P < 0.001). When the degree of resolution was reduced from 0.01 to 1.0 mm2, CV decreased by 75% in the normoxic and by 69% in the hypoxic state. Thus we conclude that 1) the decrease in PaO2 increases similarity of blood flows in nearby regions and decreases myocardial blood flow heterogeneity, and 2) similarity of regional blood flows increases with depth of the left ventricle in the normoxic state, but this transmural difference disappears in the hypoxic state.

We examined the effect of nitroglycerin (NTG) on the diameter and diastolic-to-systolic pulsation amplitude of large (ID > 100 microns) and small (ID < 100 microns) subendocardial arterioles with their segmental responses. In 10 open-chest, anesthetized pigs, subendocardial arterioles of beating hearts (n = 18) were videorecorded using a needle-probe videomicroscope. Subendocardial arteriolar diameter was determined before and 1-2 min after NTG administration (25 micrograms/kg i.v.). In an additional experiment using three pigs, we monitored the transient response of subendocardial small arterioles (n = 5) from the time before NTG administration until 3 min after NTG. NTG dilated large subendocardial arterioles by 13 +/- 4% (means +/- SD, n = 10, P < 0.001) at about 1.5 min after NTG, but not small subendocardial arterioles (2 +/- 2%, n = 8, not significant). However, the small arterioles responded transiently to NTG in an earlier phase, especially to a higher dose. The percent diameter change of large subendocardial arterioles between diastole and systole at 1.5 min after NTG administration was 32 +/- 4%, which was larger than that under control conditions (19 +/- 8%, P < 0.05). The pulsation amplitude of small subendocardial arterioles at this time was almost unchanged by NTG (16 +/- 8 vs. 17 +/- 6%, NS) but increased transiently in an earlier phase. In conclusion, NTG dilated large subendocardial arterioles on the plateau phase of its impulse (intravenously) response (approximately 1.5 min after NTG).(ABSTRACT TRUNCATED AT 250 WORDS)

To analyze the effect of coronary stenosis on the phasic pattern of blood flow velocity into the myocardium, we measured septal artery blood velocities under control conditions, i.e., no stenosis (N), with moderate (S1) and severe (S2) proximal coronary arterial stenosis, and during complete occlusion. In eight anesthetized open-chest dogs, the blood velocity measurements were performed using our 20-MHz multichannel pulsed Doppler method before and after intracoronary adenosine administration. Under N, the coronary perfusion pressure was set to approximately 100 mmHg, with S1 the poststenotic pressure was approximately 60 mmHg, and with S2 it was decreased to 35 mmHg. Septal arterial blood velocity area, which is the time-integrated envelope of the velocity, was divided into three components; systolic retrograde velocity area, systolic anterograde velocity area, and diastolic anterograde velocity area. There were no significant changes from N to S1 in any of the three velocity areas, which is a measure of the flow. Total inflow decreased from N to S2 by 52% (P less than 0.05). Diastolic anterograde flow decreased from N to S2 by 32% (P less than 0.05). Systolic retrograde flow increased from N to S2 by 75% (P less than 0.05). Systolic anterograde flow was almost unchanged by increasing stenosis. After complete occlusion, the septal artery exhibited a to-and-fro flow pattern. Adenosine-induced vasodilation reduced poststenotic pressure (P less than 0.05) and enhanced systolic retrograde flow without a significant increase in diastolic anterograde flow. Systolic retrograde flow increased at S1 by 49% (P less than 0.05) and at S2 by 27% (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

The influence of the filling condition of the unstressed volume (UV) of intramyocardial vessels on the diastolic coronary arterial pressure-flow relationship was analyzed. UV is defined as the blood volume at zero transmural pressure. In seven anesthetized, paced dogs with induced heart block, coronary artery inflow was occluded so that blood in the UV was displaced into the coronary vein by myocardial contraction. After pacing was turned off, coronary perfusion pressure was increased stepwise to seven target pressures (20-90 mmHg). After reperfusion, left anterior descending coronary arterial (LAD) flow reached an initial quasi-steady level, but great cardiac venous (GCV) flow was absent (UV-unfilled phase). With reappearance of the GCV flow (UV-filled phase), LAD flow decreased to a final steady level. Pressure-flow relationships during both phases were linear (r = 0.97-0.99). The inverse of the slope of the pressure-flow relationship during the UV-filled phase, 0.69 +/- 0.08 mmHg.min.ml-1, was significantly higher than that during the UV-unfilled phase, 0.55 +/- 0.06 mmHg.min.ml-1 (P less than 0.01), although the zero-flow pressure intercepts were similar (18.9 +/- 1.8 mmHg for UV filled vs. 19.9 +/- 2.2 for UV unfilled). These results indicate that diastolic coronary arterial inflow is impeded by a blood volume within intramyocardial vessels that exceeds the UV.

To evaluate regional myocardial function, we developed a curvature sensor for direct and instantaneous measurement of the regional curvature radius of the LV wall. The sensor is a bimorph of two sheets of thin piezoelectric polymer film. The relation between output voltage of the sensor and the reciprocal of the known curvature radius has been shown to be linear. In anesthetized dogs, we inserted the curvature sensor into the subepimyocardium with a specially designed introducer. Special care was taken to insert it parallel to the epicardial surface. Circumferential regional curvature radius, which is defined as the reciprocal of regional curvature, showed the same phasic changes as the short-axis diameter under control conditions. Under regional ischemia caused by transient occlusion of the coronary artery, amplitude of the phasic change in regional curvature radius decreased, whereas that of the short-axis diameter did not change or slightly increased. Phasic changes in regional curvature radius cannot be estimated from short-axis diameter during regional ischemia. We calculated regional wall tension from the directly measured regional curvature radius and LV pressure and found that tension-length loop clearly differentiates between regional myocardial function under control and ischemic conditions. We have concluded that our newly developed curvature sensor is accurate, that its practical use is feasible, and that measurement of the regional curvature radius of the LV wall provides detailed and accurate information on regional shape and function of the left ventricle.