Effect of Treatment on Ventricular Function and Troponin I Proteolysis in Reperfused Myocardium

Abstract

Effects of ischemia time and treatment interventions upon troponin I (TnI) proteolysis and function of reperfused myocardium were examined in isolated, perfused rabbit hearts. Hearts were randomized to 90 min aerobic perfusion, 15 min low-flow (1 ml/min) ischemia (I) and 60 min reperfusion (R) or 60 min low-flow I and 60 min R. Hearts subject to 60 min I and 60 min R received either no treatment, l -arginine treatment, or treatment with oxygen free radical (OFR) scavengers (mercapto-proponyl-glycine, catalase and superoxide dismutase). Hearts from cholesterol-fed rabbits were also studied after 60 min I and R. Isovolumic LV pressure and heart rate were recorded throughout and Western analysis of ventricular myocardium, using 3 specific antibodies, detected intact TnI (29 kDa) and TnI fragment (25 kDa). Hearts subject to 15 min I had minimal irreversible injury (TTC negative region=0.6±0.4% LV) but hearts subject to 60 min I had more extensive injury (TTC negative=40.7±5.8% LV). Recovery of rate–pressure product after 15 min I and 60 min R (56±9% of baseline) was better than after 60 min I and 60 min R (23±9%, P<0.01). Bothl -arginine and OFR scavengers were associated with better recovery of function after 60 min I, (66±7% and 72±3% of baseline respectively, P<0.01 v no treatment) but cholesterol hearts had poor recovery after 60 min I (37±8%). The 25 kDa TnI (% total TnI immunoreactivity) was 8.7±0.9% in controls, 10.0±1.6% after 15 min I and 60 min R, and 17.4±2.4% after 60 min I and 60 min R (P<0.01v controls and 15 min I). The proportion of 25 kDa TnI was increased in all hearts after 60 min I and did not change with treatment (l -arginine 16.8±1.8%, OFR scavengers 16.0±3.2%, cholesterol 14.0±1.9%). There was no relation between proportion of 25 kDa TnI and recovery of function. Samples from freshly excised rabbit hearts and human right atria also had 25 kDa TnI (relative intensities 8.5±2.3% and 5.1±2.6% respectively). Although TnI fragmentation increases after prolonged ischemia and reperfusion, the functional recovery of stunned myocardium is independent of degree of TnI fragmentation.