title: Helium postconditioning regulates expression of caveolin-1 and -3 and induces RISK pathway activation after ischaemia/reperfusion in cardiac tissue of rats.
abstract: Caveolae, lipid enriched invaginations of the plasma membrane, are epicentres of cellular signal transduction. The structural proteins of caveolae, caveolins, regulate effector pathways in anaesthetic-induced cardioprotection, including the RISK pathway. Helium (He) postconditioning (HePoc) is known to mimic anaesthetic conditioning and to prevent damage from myocardial infarction. We hypothesize that HePoc regulates caveolin-1 and caveolin-3 (Cav-1 and Cav-3) expression in the rat heart and activates the RISK pathway. Male Wistar rats (n=8, each group) were subjected to 25min of cardiac ischaemia followed by reperfusion (I/R) for 5, 15 or 30min (I/R 5/15/30). The HePoc groups underwent I/R with 70% helium ventilation during reperfusion (IR+He 5/15/30min). Sham animals received surgical treatment without I/R. After each protocol blood and hearts were retrieved. Tissue was obtained from the area-at-risk (AAR) and non-area-at-risk (NAAR) and processed for western blot analyses and reverse-transcription-real-time-polymerase-chain-reaction (RT-qPCR). Protein analyses revealed increased amounts of Cav-1 and Cav-3 in the membrane of I/R+He15 (AAR: Cav-1, P<0.05; Cav-3, P<0.05; both vs. I/R15). In serum, Cav-3 was found to be elevated in I/R+He15 (P<0.05 vs. I/R15). RT-qPCR showed increased expression of Cav-1 in IR+He15 in AAR tissue (P<0.05 vs. I/R15). Phosphorylation of RISK pathway proteins pERK1/2 (AAR: P<0.05 vs. I/R15) and pAKT (AAR: P<0.05; NAAR P<0.05; both vs. I/R15) was elevated in the cytosolic fraction of I/R+He15. These results suggest that 15min of HePoc regulates Cav-1 and Cav-3 and activates RISK pathway kinases ERK1/2 and AKT. These processes might be crucially involved in HePoc mediated cardioprotection.
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands; Department of Anaesthesiology and Intensive Care Medicine, University Hospital Schleswig-Holstein, Campus Kiel, Arnold-Heller-Straße 3, 24105 Kiel, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Flick
authors.affiliation: Department of Anaesthesiology and Intensive Care Medicine, University Hospital Schleswig-Holstein, Campus Kiel, Arnold-Heller-Straße 3, 24105 Kiel, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Albrecht
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Gezina T M L
authors.initials: GTML
authors.lastname: Oei
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Renske
authors.initials: R
authors.lastname: Steenstra
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Raphaela P
authors.initials: RP
authors.lastname: Kerindongo
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Coert J
authors.initials: CJ
authors.lastname: Zuurbier
authors.affiliation: Veterans Affairs San Diego Healthcare System and Department of Anaesthesiology, University of California, San Diego, 9500 Gilman Drive, 92093 La Jolla, California, USA.
authors.forename: Hemal H
authors.initials: HH
authors.lastname: Patel
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Markus W
authors.initials: MW
authors.lastname: Hollmann
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Preckel
authors.affiliation: Department of Anaesthesiology, Laboratory of Experimental Intensive Care and Anaesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands. Electronic address: N.C.Hauck@amc.uva.nl.
authors.forename: Nina C
authors.initials: NC
authors.lastname: Weber
mesh_terms: D000818:Animals; D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism; D051244:Caveolin 3 / Q000235:genetics / Q000378:metabolism; D022461:Caveolins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006371:Helium / Q000494:pharmacology*; D057775:Ischemic Postconditioning; D008297:Male; D015428:Myocardial Reperfusion Injury / Q000378:metabolism / Q000473:pathology*; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D011494:Protein Kinases / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D051242:Caveolin 1; D051244:Caveolin 3; D022461:Caveolins; D012333:RNA, Messenger; D006371:Helium; D011494:Protein Kinases
keywords: Cardiac postconditioning; Caveolin; Helium; Ischaemia reperfusion; Noble gases; RISK pathway