Signaling pathways involved in postconditioning-induced cardioprotection of human myocardium, in vitro

We examined the respective role and relationship between protein kinase C (PKC), mitochondrial adenosine triphosphate-sensitive potassium (mitoK(ATP)) channel and p38 mitogen-activated protein kinase (MAPK) in postconditioning of human myocardium, in vitro. Isometrically contracting, isolated human right atrial trabeculae were exposed to 30 min hypoxia and 60 min reoxygenation. Phorbol 12-myristate 13-acetate (a PKC activator), diazoxide (a mitoK(ATP) opener) and anisomycin (a p38 MAPK activator) were superfused in early reoxygenation alone and with calphostin C (a PKC inhibitor), 5-hydroxy-decanoate (5-HD, a mitoK(ATP) channel inhibitor) and SB 202190 (a p38 MAPK inhibitor). Developed force at the end of the 60 min reoxygenation (FoC(60)) period was compared between groups (mean +/- SD). Phorbol 12-myristate 13-acetate (91 +/- 4% of baseline), diazoxide (85 +/- 5% of baseline) and anisomycin (90 +/- 4% of baseline) enhanced the FoC(60) as compared with the control group (53 +/- 7% of baseline, P < 0.0001). The enhanced FoC(60) induced by phorbol 12-myristate 13-acetate was abolished by calphostin C (52 +/- 5% of baseline) and 5-HD (56 +/- 3% of baseline), but not by SB 202190 (90 +/- 8%). The diazoxide-induced recovery of FoC(60) was attenuated by 5-HD (55 +/- 6% of baseline), but was not modified by calphostin C (87 +/- 5% of baseline) and SB 202190 (90 +/- 8% of baseline). The anisomycin-induced recovery of FoC(60) was abolished by calphostin C (61 +/- 9% of baseline) and SB 202190 (52 +/- 8% of baseline), but not by 5-HD (88 +/- 6% of baseline). In conclusion, PKC activation, opening of mitoK(ATP) channels and p38 MAPK activation in early reoxygenation induced the postconditioning of human myocardium, in vitro. Furthermore, PKC activation was upstream of the opening of mitoK(ATP) channels; p38 MAPK acted on PKC. Therefore, mitoK(ATP) and p38 MAPK seemed to be involved in two independent pathways

Sevoflurane- and desflurane-induced human myocardial post-conditioning through Phosphatidylinositol-3-kinase/Akt signalling

BACKGROUND: The role of phosphatidylinositol-3-kinase (PI3K) in sevoflurane- and desflurane-induced myocardial post-conditioning remains unknown. METHODS: We recorded isometric contraction of isolated human right atrial trabeculae (oxygenated Tyrode's at 34 degrees C, stimulation frequency 1 Hz). In all groups, a 30-min hypoxic period was followed by a 60-min reoxygenation period. At the onset of reoxygenation, muscles were exposed to 5 min of sevoflurane 1%, 2%, and 3%, and desflurane 3%, 6%, and 9%. In separate groups, sevoflurane 2% and desflurane 6% were administered in the presence of 100 nM wortmannin, a PI3K inhibitor. Recovery of force after the 60-min reoxygenation period was compared between groups (mean +/- SD). RESULT: As compared with the Control group (49 +/- 7% of baseline) PostC by sevoflurane 1%, 2%, and 3% (78 +/- 4%, 79 +/- 5%, and 85 +/- 4% of baseline, respectively) and desflurane 3%, 6%, and 9% (74 +/- 5%, 84 +/- 4%, and 86 +/- 11% of baseline, respectively) enhanced the recovery of force. This effect was abolished in the presence of wortmannin (56 +/- 5% of baseline for sevoflurane 2%+wortmannin; 56 +/- 3% of baseline for desflurane 6%+wortmannin). Wortmannin alone had no effect on the recovery of force (57 +/- 7% of baseline). CONCLUSION: In vitro, sevoflurane and desflurane post-conditioned human myocardium against hypoxia through activation of phosphatidylinositol-3-kinase

The inotropic and lusitropic effects of ketamine in isolated human atrial myocardium: The effect of adrenoceptor blockade

We studied the direct myocardial effects of racemic ketamine, in the presence of \u3b1- and \u3b2-adrenoceptor blockade, on isolated human right atrial myocardium. Isometric force of contraction (FoC), its first derivative with time (+dF/dt), the contraction relaxation coupling parameter R2 = (+dF/dt) / (-dF/dt), and time to half relaxation (T1/2) were recorded before and after addition of 10-6, 10-5 and 10-4 M racemic ketamine alone and in the presence of \u3b1-adrenoceptor blockade (phentolamine 10-6 M) and \u3b2-adrenoceptor blockade (propranolol at 10-6 M). Ketamine had a moderate positive inotropic effect at 10-5 M (FoC, 104% \ub1 5% of baseline value; P = 0.03) and 10-4 M (FoC, 107% \ub1 11% of baseline value; P = 0.09). Racemic ketamine had a negative inotropic effect in the presence of propranolol (FoC, ketamine 10-6 M, 77% \ub1 11%; ketamine 10-5 M, 63% \ub1 16%; ketamine 10-4 M, 62% \ub1 17% of baseline; P < 0.001) but not phentolamine (FoC, ketamine at 10-6 M, 94% \ub1 6%; ketamine 10-5 M, 96% \ub1 5%; and ketamine 10-4 M, 98% \ub1 15% of baseline). Ketamine decreased T1/2 (ketamine 10-5 M, 94% \ub1 3% of baseline value; P < 0.001 and ketamine 10-4 M, 90% \ub1 9% of baseline value; P = 0.007) but did not modify R2. In human right atrial myocardium, racemic ketamine induced a moderate positive inotropic effect and hastened isometric relaxation. In the presence of \u3b2-adrenoceptor blockade it induced a direct negative inotropic effect

Role of 70-kDa ribosomal protein S6 kinase, nitric oxide synthase, glycogen synthase kinase-3 beta, and mitochondrial permeability transition pore in desflurane-induced postconditioning in isolated human right atria

Desflurane during early reperfusion has been shown to postcondition human myocardium. Whether it involves "reperfusion injury salvage kinase" pathway remains incompletely studied. The authors tested the involvement of 70-kDa ribosomal protein S6 kinase, nitric oxide synthase, glycogen synthase kinase (GSK)-3beta, and mitochondrial permeability transition pore in desflurane-induced postconditioning

Desflurane-induced postconditioning of diabetic human right atrial myocardium in vitro

AIM: We tested the hypothesis that brief exposure to desflurane at the time of reoxygenation might be able to protect against hypoxia-reoxygenation injury in human myocardium from diabetic (insulin-dependent, ID; and non-insulin-dependent, NID) patients and non-diabetic (ND) subjects. METHODS: The force of contraction (34 degrees C, stimulation frequency 1Hz) in the right atrial trabeculae was recorded during 30min of hypoxia followed by 60min of reoxygenation. Desflurane (at 3, 6 and 9%) was administered during the first 5min of reoxygenation. The force of contraction at the end of the 60-min reoxygenation period (FoC(60)) was compared in the study groups (means+/-SD). RESULTS: In the ND group, desflurane at 3, 6 and 9% (FoC(60): respectively 78+/-10%, 84+/-4% and 85+/-12% of baseline) enhanced the recovery of FoC(60) compared with the ND-controls (53+/-7% of baseline; P<0.05). In the ID group, desflurane at 3% (61+/-4%) did not modify the recovery of FoC(60) compared with the ID-controls (54+/-6%), whereas desflurane at 6 and 9% (75+/-11% and 81+/-8%, respectively) enhanced the recovery of FoC(60)vs the controls (P<0.05). In the NID group, desflurane at 3% (57+/-5%) also failed to modify the recovery of FoC(60) compared with the NID-controls (52+/-10%), while desflurane at 6 and 9% (80+/-10% and 79+/-7%, respectively) enhanced the recovery of FoC(60)vs the controls (P<0.05). CONCLUSION: Desflurane in vitro was able to postcondition diabetic (both ID and NID) human myocardium at 6 and 9%, but not at 3%

Etomidate has no effect on hypoxia reoxygenation and hypoxic preconditioning in isolated human right atrial myocardium

BACKGROUND: We examined the effects of etomidate on recovery of contractile function after hypoxia reoxygenation and hypoxic preconditioning in vitro using isolated human myocardium. METHODS: Human right atrial myocardium were obtained at the time of cardiac surgery from 38 adults patients. We recorded isometric force of contraction (FoC) of atrial trabeculae suspended in an oxygenated Tyrode's solution (34 degrees C, stimulation frequency 1 Hz). In all groups, a 30-min hypoxic period was followed by 60 min of reoxygenation (HR). In separate groups, muscles were exposed to etomidate (10(-7), 10(-6), 10(-5) M) 10 min before and throughout the HR periods. Hypoxic preconditioning was induced by 4-min hypoxia followed by 7-min reoxygenation applied before HR periods. Etomidate 10(-5) M was administered before, throughout, and after the hypoxic preconditioning stimulus. Recovery of FoC (expressed as % of baseline value) at the end of HR was compared among groups. RESULTS: Compared with the control group (FoC: 52%+/-10%), etomidate 10(-7) M (FoC: 57%+/-9%; P=0.24), 10(-6) M (FoC: 61%+/-11%; P=0.10), and 10(-5) M (FoC: 54%+/-9%; P=0.29) did not modify the recovery of FoC after HR. Hypoxic preconditioning-induced increase in the recovery of FoC (87%+/-5%; P<0.001 vs control group) was not modified in the presence of etomidate 10(-5) M (FoC: 86%+/-7%; P=0.74 vs hypoxic preconditioning group). CONCLUSIONS: Etomidate did not modify the in vitro FoC of human myocardium exposed to HR. Furthermore, etomidate did not modify the protective effect of hypoxic preconditioning