1H NMR Detection of Lactate and Alanine in Perfused Rat Hearts during Global and Low Pressure Ischemia

A spin-echo method is presented for obtaining high resolution, 13C coupled, proton spectra of lactate and alanine in intact, beating rat hearts. All hearts were depleted of glycogen prior to prolonged perfusion with either 10 mM unenriched glucose or [1-13C]glucose to restore glycogen. These two groups of hearts were then examined by 1H NMR during prolonged global (zero flow) or low pressure (low flow) ischemia. During global ischemia, lactate was derived from both glucose and glycogen, with endogenous glycogen contributing twice as much lactate as exogenous glucose. During low perfusion pressure ischemia, however, lactate was derived exclusively from exogenous glucose. The entire pool of lactate (both 12C and 13C) was visible by NMR in intact, glucose perfused hearts while alanine was not detected. However, upon adding 10 mM pyruvate to the perfusate, the entire alanine pool became NMR visible while some of the lactate became NMR invisible. These observations indicate that the NMR visibility of small, usually highly mobile metabolites such as alanine and lactate is not always 100% in intact hearts and that the NMR visibility of these molecules may depend upon which exogenous substrate is presented to the heart.