OBJECTIVE: We demonstrated earlier that postischemic addition of insulin improves recovery of function in isolated rat heart by phosphatidylinositol 3-kinase. Activation of phosphatidylinositol 3-kinase before ischemia improves recovery of the heart after ischemia through protein kinase C. We tested whether protein kinase C activation is required for the positive inotropic effect of insulin during reperfusion. METHODS: Isolated working rat hearts were perfused with Krebs-Henseleit buffer containing [2-(3)H]glucose (5 mmol/L, 0.05 microCi/mL) plus oleate (0.4 mmol/L) and were subjected to 15 minutes of global ischemia followed by 35 minutes of reperfusion with or without insulin (1 mU/mL). We measured cardiac power, glucose uptake, and tissue metabolites. The protein kinase C inhibitor chelerythrine (5 micromol/L) was added either at the beginning of the experiment or together with insulin. Experiments were repeated under normoxic conditions. RESULTS: Cardiac power before ischemia was 9.63 to 12.4 mW. Insulin improved recovery of power after ischemia (96.3% +/- 10.8% versus 65.7% +/- 3.79%, P <.05). This effect was abolished by chelerythrine (55.3% +/- 6.49%). However, chelerythrine given at reperfusion did not block insulin's effect on recovery (101.0% +/- 4.25%, P <.05). Postischemic glucose uptake was not increased by insulin (3.07 +/- 0.32 before, 3.45 +/- 0.34 micromol/min/gdw after ischemia, not significant) and was not affected by chelerythrine (3.01 +/- 0.26 before, 3.29 +/- 0.32 micromol/min/gdw after ischemia, not significant). Under normoxic conditions, chelerythrine did not influence insulin's effects on glucose uptake or power. CONCLUSION: The results suggest that (1) insulin's effect on recovery is dependent on ischemia-induced protein kinase C activation, (2) the activity of protein kinase C during reperfusion may not be important for this effect of insulin, and (3) protein kinase C plays no role in insulin's effect on glucose uptake under normoxic or postischemic conditions.
OBJECTIVE: We demonstrated earlier that postischemic addition of insulin improves recovery of function in isolated rat heart by phosphatidylinositol 3-kinase. Activation of phosphatidylinositol 3-kinase before ischemia improves recovery of the heart after ischemia through protein kinase C. We tested whether protein kinase C activation is required for the positive inotropic effect of insulin during reperfusion. METHODS: Isolated working rat hearts were perfused with Krebs-Henseleit buffer containing [2-(3)H]glucose (5 mmol/L, 0.05 microCi/mL) plus oleate (0.4 mmol/L) and were subjected to 15 minutes of global ischemia followed by 35 minutes of reperfusion with or without insulin (1 mU/mL). We measured cardiac power, glucose uptake, and tissue metabolites. The protein kinase C inhibitor chelerythrine (5 micromol/L) was added either at the beginning of the experiment or together with insulin. Experiments were repeated under normoxic conditions. RESULTS: Cardiac power before ischemia was 9.63 to 12.4 mW. Insulin improved recovery of power after ischemia (96.3% +/- 10.8% versus 65.7% +/- 3.79%, P <.05). This effect was abolished by chelerythrine (55.3% +/- 6.49%). However, chelerythrine given at reperfusion did not block insulin's effect on recovery (101.0% +/- 4.25%, P <.05). Postischemic glucose uptake was not increased by insulin (3.07 +/- 0.32 before, 3.45 +/- 0.34 micromol/min/gdw after ischemia, not significant) and was not affected by chelerythrine (3.01 +/- 0.26 before, 3.29 +/- 0.32 micromol/min/gdw after ischemia, not significant). Under normoxic conditions, chelerythrine did not influence insulin's effects on glucose uptake or power. CONCLUSION: The results suggest that (1) insulin's effect on recovery is dependent on ischemia-induced protein kinase C activation, (2) the activity of protein kinase C during reperfusion may not be important for this effect of insulin, and (3) protein kinase C plays no role in insulin's effect on glucose uptake under normoxic or postischemic conditions.
Authors: Michael Schwarzer; Steven L Britton; Lauren G Koch; Ulrik Wisloff; Torsten Doenst Journal: Basic Res Cardiol Date: 2010-02-05 Impact factor: 17.165