Abstract
Real‐time measurement of nitric oxide (NO) before, during and after study interventions has numerous advantages over multiple endpoint measurements at various times. However, no reliable method has been described in Langendorff‐prepared hearts. Rat isolated hearts were loaded for 30 min with 20 µM of the fluorescent dye 4,5‐diaminofluorescein (DAF‐2) whose fluorescence (F) increases irreversibly in the presence of NO. Changes in NO were provoked by any combination of 30/120 min ischemia/reperfusion, the NO donor sodium nitroprusside and/or its scavenger cPTIO. Using a bifurcated fiberoptic probe left ventricular epicardial DAF‐2 F was measured online at 490/540 nm excitation/emission. Multiple regression analysis (alpha .05) was used to compare epicardial F after 185 min perfusion to confocal microscopy (488/550 nm) of transverse slices and to high‐performance liquid chromatography of tissue homogenate. Overall, all three measurements correlated significantly to each other and changed adequately with above treatments. Although online fluorescence lacks the spatial resolution of confocal microscopy it may provide a unique insight into real‐time effects of IR and potential cardioprotective treatments on the production of NO. This may serve as an important adjunct tool to investigate its role in studying cardioprotective strategies in isolated hearts. Supported by Department of Veterans Affairs (CARA‐026‐10F) and NIH.Grant Funding Source: Supported by Department of Veterans Affairs and NIH.
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