Physiologisches Institut Reactive oxygen species: Importance for ischemia/reperfusion (injury) Prof. Dr. Rainer Schulz
Reactive oxygen species (ROS) in ischemia/reperfusion injury (IRI) ROS GOOD: Endogenous protection BAD: Infarction ROS
Potential sources of ROS Cardiomyocytes Other cell types p66 SOD2 Mitochondria CAT SOD1 Giorgio et al., Nat Rev Mol Cell Biol, 8:722-728, 2007; Akki et al., J Mol Cell Cardiol 47: 15 22, 2009
Formation of ROS during ischemia Rat cardiomyocytes ROS generation: amytal (2.5 mm, a site I electron transport inhibitor), rotenone (10 μm, a site I electron transport inhibitor), bovine ROS formation during liver SOD (200 U/ml, dismutates superoxide to H2O2), (low-flow) diethyldithiocarbamic ischemia acid (DDC, 1 mm, which appears chelates to Cu be in Cu,Zn-SOD, primarily thereby inhibiting its function), nitro-larginine methyl caused ester [L-NAME, by mitochondria! 200 μm, inhibitor of nitric oxide synthase (NOS)], apocynin (300 μm, an inhibitor of NADPH oxidases). Cytosolic SOD inhibitor Complex I inhibitors NOX inhibitor NOS inhibitor Becker et al., Am J Physiol 277 Heart Circ Physiol 46: H2240 H2246, 1999
Dog Formation of ROS during ischemia/reperfusion Reperfusion Ischemia Bolli et al., Proc Natl Acad Sci USA 86: 4695-4699, 1989
ROS during IRI contribute to cellular damage contribute to cellular protection Function Morphology Function Morphology (Postischemic Reperfusion, Heart Failure) (Ischemia/Reperfusion Injury)
Mice NOX2 and irreversible IRI NOX2-KO Hoffmeyer et al., Circ Res 87: 812-817, 2000
Mice Catalase (CAT), superoxid dismutase (SOD) and irreversible IRI Cardiomyocytes Cardiomyocytes SOD2 Mitochondria CAT SOD1 Giorgio et al., Nat Rev Mol Cell Biol, 8:722-728, 2007 Loor et al., Biochem Biophys Acta 2011
Mice GSH during ischemia/reperfusion GSH Mitochondria
GSH and irreversible IRI Pig Kupatt et al., Cardiovasc Res 61: 530-537, 2004
Aldolase reductase and irreversible IRI
ROS and mitochondrial morphology Cell (Cardio)- protection mrna BBA 2011
Mice ROS and mitochondrial morphology Loor et al., Biochem Biophys Acta 2011
ROS and mitochondrial morphology Kuznetsova et al, Int J Biochem & Cell Biol 41: 1928 1939, 2009
OPA1, mitochondrial morphology and IRI Chen et al., Cardiovasc Res 84: 91-99, 2009
ROS during IRI and protection from it contribute to cellular damage contribute to cellular protection Function Morphology Function Morphology (Postischemic Reperfusion, Heart Failure) (Ischemia/Reperfusion Injury)
Ischemic pre- and post-conditioning Dog Coronary occlusion 30 Infarct size (% area at risk) Control Precon (Ischemia/ Drugs) 60 min 5 10 min 3h 3h 25 20 15 10 P<0.05 Postcon 3h 5 0 control Precon Postcon Zhao et al., Am J Physiol 285: H579-H588, 2004
ROS and ischemic preconditioning Garciarena et al., J Appl Physiol 105: 1706-1713, 2008
NOX2, ROS and ischemic preconditioning Mice NOX2-KO Bell et al., FASEB J 2005
Cylophilin D, ROS and ischemic preconditioning
Pre- and Post-conditioning: Signal transduction adenosine bradykinin opioids, UCN ANP BNP adenosine bradykinin opioids, UCN IGF-1 FGF-2 IL-6 type cytokines NO TNFα GPCR PI3K Akt NPR pgc GPCR NPR gp130 JAK TNF-R CB-R adiponectin only adenosine enos NO H11K PI3K Akt ERK STAT3 AMPK NO p38 sgc PKG enos NO P70S6K SIRT1 inos MnSOD aldose reductase ROS MPTP PKC MPTP GSK3β mitochondrium nucleus K ATP mitochondrium Cx43 mitochondrium : age-dependent : species-dependent Heusch, Boengler, Schulz, Circulation 118: 1915-1919, 2008; Boengler, Schulz, Heusch, Cardiovasc Res 2009
Heinzel et al., Circ Res 97:583-586, 2005 Connexin 43 (Cx43), ROS and ischemic preconditioning Knockout mice MTR fluorescence [% above control] 80 70 Infarct size (% area at risk) WT Cx43 +/ 60% 60 50% 40% 30% 20% 10% p<0.05 50 40 30 20 * * 0% WT Cx43 +/ Cx43 +/ Diazoxide Menadione 10 0 PLA Diaz PLA Diaz Mena
Rat heart in vitro Mitochondrial uncoupling, ROS and IRI Yue et al., Am J Physiol 281: H590-H595, 2001 Minners et al., Cardiovasc Res 47: 68-73, 2000
ROS scavenging and ischemic preconditioning 35 Infarct size [% area at risk] 30 25 20 15 10 5 P<0.05 0 Placebo IP10 Ascorbic acid IP10 + Ascorbic acid
Ovize et al, Cardiovasc Res 87: 406-423, 2010 Ischemic Postconditioning: Signal transduction MKP-1
Tsutsumi et al., Life Sci 81: 1223-1227, 2007 Mice ROS scavenging and ischemic postconditioning
ROS during IRI and protection from it contribute to cellular damage contribute to cellular protection Function Morphology Function Morphology (Postischemic Reperfusion, Heart Failure) (Ischemia/Reperfusion Injury)