Mechanisms involved in the desflurane-induced post-conditioning of isolated human right atria from patients with type 2 diabetes  S. Lemoine, L. Zhu, C. Buléon, M. Massetti, J-L. Gérard, P. Galera, J-L. Hanouz  British Journal of Anaesthesia  Volume 107, Issue 4, Pages 510-518 (October 2011) DOI: 10.1093/bja/aer201 Copyright © 2011 The Author(s) Terms and Conditions

Fig 1 Schematic diagram depicting the experimental protocol. (a) Contracting muscle experimental protocols. In the Diab-Des+Cal and Diab-Cal groups, calphostin C was administered at 1 µM. In the Diab-Des+5-HD and Diab-5-HD groups, 5-HD was administered at 800 µM. In the Diab-PMA and Diab-Diazoxide groups, PMA was administered at 1 µM and diazoxide was administered at 100 µM, respectively. (b) Western blot experimental protocols. Cal, calphostin C; Des, desflurane; Diab, diabetic; 5-HD, 5-hydroxydecanoate; PMA, phorbol 12-myristate 13-acetate. British Journal of Anaesthesia 2011 107, 510-518DOI: (10.1093/bja/aer201) Copyright © 2011 The Author(s) Terms and Conditions

Fig 2 Recovery of the FoC of isolated human right atrial trabeculae at the end of the 60 min reoxygenation period after the 30 min hypoxic challenge in groups exposed to desflurane (6%) (Diab-Des) alone or in the presence of calphostin C (Diab-Des+Cal) and 5-HD (Diab-Des+5-HD). Effect of the administration of PMA (Diab-PMA) and diazoxide (Diab-Diazoxide) at the beginning of reoxygenation on the time course of the FoC (expressed as a percentage of baseline) of the isolated human right atrial trabeculae during a 30 min hypoxic challenge followed by a 60 min reoxygenation period. Data are mean (sd). *P<0.001 vs the Diab-Control, Diab-Des+Cal, Diab-Des+5-HD, Diab-Cal, and Diab-5-HD groups. Cal, calphostin C; Des, desflurane; Diab, diabetic; 5-HD, 5-hydroxydecanoate; PMA, phorbol 12-myristate 13-acetate. British Journal of Anaesthesia 2011 107, 510-518DOI: (10.1093/bja/aer201) Copyright © 2011 The Author(s) Terms and Conditions

Fig 3 Western blot analysis showing levels of total Akt, phosphorylated Akt (a), total glycogen synthase kinase 3β (GSK-3β total), and phosphorylated GSK-3β (Ser9) (phospho-GSK-3β) (b) after 5 min of reoxygenation alone (Control) or in the presence of 5 min of 6% desflurane (Des). Equal loading was confirmed by western blot with an anti-β-tubulin antibody. The relative total Akt, phospho-Akt, total GSK-3β, and phospho-GSK-3β protein levels were calculated by averaging the results obtained from four independent experiments. Example blot shown is a representative image of a single blot. Data are mean (sd). *P<0.0001 vs the Control and Diab-Control groups. Des, desflurane; Diab, diabetic; GSK-3β, glycogen synthase kinase-3β. British Journal of Anaesthesia 2011 107, 510-518DOI: (10.1093/bja/aer201) Copyright © 2011 The Author(s) Terms and Conditions

Fig 4 Proposed schematic representation of the signalling pathways leading to desflurane induced post-conditioning of the human myocardium with type 2 diabetes, in vitro. Brief administration of desflurane in the early reoxygenation period induced PKC activation, the opening of mitoKATP channels, phosphorylation of Akt and GSK 3β, which mediate the cardioprotective effect. Moreover, in early reoxygenation, activation of PKC (via PMA administration) and the opening of mitoKATP channels (via diazoxide administration) induced myocardial post-conditioning. GSK-3β, glycogen synthase kinase-3β; mitoKATP, mitochondrial adenosine triphosphate-sensitive potassium channel; PKC, protein kinase C; PMA, phorbol 12-myristate 13-acetate; 5-HD, 5-hydroxydecanoate. British Journal of Anaesthesia 2011 107, 510-518DOI: (10.1093/bja/aer201) Copyright © 2011 The Author(s) Terms and Conditions