PD Dr. Kerstin Boengler
Physiologisches Institut
Oxidant stress: Consequences for
ischemia/reperfusion
ROS
ROS
Reactive oxygen species (ROS) in ischemia/reperfusion injury (IRI)
GOOD:Endogenous
protectioncontribute to
BAD:Infarction contribute to
Burns et al., J Am Coll Cardiol 39:30–36, 2002
6 Monthmortality(%)
Infarct size (% area at risk)
Myocardial infarction and prognosisPatient
Giorgio et al., Nat Rev Mol Cell Biol, 8:722-728, 2007
Potential sources of ROS
; Akki et al., J Mol Cell Cardiol 47: 15–22, 2009
Mitochondria
Other cell types
p66
SOD1
Cardiomyocytes
SOD2
CAT
Becker et al., Am J Physiol 277 Heart Circ Physiol 46: H2240–H2246, 1999
Complex I inhibitors
Formation of ROS during ischemia
Rat cardiomyocytes
NOX inhibitorNOS inhibitor
Cytosolic SOD inhibitor
ROS formation during (low-flow) ischemia
appears to be primarily caused by mitochondria!
Bolli et al., Proc Natl Acad Sci USA 86: 4695-4699, 1989
Formation of ROS during ischemia/reperfusion
Ischemia
Dog
Reperfusion
contribute to cellular protection
MorphologyFunction
contribute to cellular damage
MorphologyFunction
(Ischemia/Reperfusion Injury)(Postischemic Reperfusion,Heart Failure)
ROS during IRI
NOX2, NOX4 and irreversible IRI
Matsushima, Circ Res 2013
systemic NOX2 KO mice cardiac NOX4 KO mice
MAO
p66Shc ROSPKCβ
MPTPp66Shc
Cytc
ROSCypD
CypD
ox ox
MAO
p66Shc ROSPKCβ
MPTPp66Shc
Cytc
ROSCypD
CypD
MAOMAO
p66Shcp66Shc ROSPKCβPKCβ
MPTPp66Shcp66Shc
Cytc
ROSCypD
CypD
ox ox
Cx43
Giorgio et al., Nat Rev Mol Cell Biol, 8:722-728, 2007
Mitochondria
p66
CardiomyocytesMice
Mitochondria-derived ROS
Normoxia
0
550
600
650
700
750
800
850
900
ControlClorgyline50 mM
Fluo
resc
ence
inte
nsity
(AU
)IR Clorgyline
25 mM
ROS formation by MAO during
ischemia/reperfusion
Ischemia/Reperfusion
Clorgyline 50 mM
Clorgyline 25 mM
Rat
Mitochondria-derived ROS
Fabio di Lisa
RV
WT
liver
WT
liver
p66S
hc-/-
RV
p66
Shc-
/-
p66
p52p46
Ponceau S
A
B
GAPDH
MnSOD
HDAC2
Serca2a
Na+/K+-ATPase
24 kDa
36 kDa
70 kDa
110 kDa
110 kDa
RV
cont
rol
H2O
2
mito
p66p52p46
RV
cont
rol
H2O
2MnSOD24 kDa
50 kDa
mitoC
RV
WT
liver
WT
liver
p66S
hc-/-
RV
p66
Shc-
/-
p66
p52p46
Ponceau S
A
RV
WT
liver
WT
liver
p66S
hc-/-
RV
p66
Shc-
/-
p66
p52p46
Ponceau S
A
B
GAPDH
MnSOD
HDAC2
Serca2a
Na+/K+-ATPase
24 kDa
36 kDa
70 kDa
110 kDa
110 kDa
RV
cont
rol
H2O
2
mito
p66p52p46
RV
cont
rol
H2O
2MnSOD24 kDa
50 kDa
mitoC
MiceStress-induced p66Shc translocation
Mitochondria
p66
*P<0.05
0
10
20
30
40
50
60
70
80
90
-/-0
10
20
30
40
50
60
70
80
90
Wildtyp p66shc -/-
Infarct size (% area at risk)
Mice
p66shc and irreversible IRI
Giorgio et al., Nat Rev Mol Cell Biol, 8:722-728, 2007
Mitochondria
SOD1
Cardiomyocytes
SOD2
CAT
Loor et al., Biochem Biophys Acta 2011
Cardiomyocytes
Mice
Catalase (CAT), superoxide dismutase (SOD) and irreversible IRI
Mice
Mitochondrial fission and IRI
Mdivi: mitochondrial division inhibitor
Cel
ls w
ith e
long
ated
mito
chon
dria
(%)
Vector control Mfn1 Mfn2 DrpK38A hFis1
Ong et al., Circulation 2010
contribute to cellular protection
MorphologyFunction
contribute to cellular damage
MorphologyFunction
(Ischemia/Reperfusion Injury)(Postischemic Reperfusion,Heart Failure)
ROS during IRI and protection from it
Coronary occlusion
Control
Precon(Ischemia/Drugs)
60 min
3h5 10 min
3h
Infarct size(% area at risk)
control Precon
30
25
10
20
15
5
0
P<0.05
Ischemic preconditioning
Zhao et al., Am J Physiol 285: H579-H588, 2004
Dog
Heusch, Boengler, Schulz, Circulation 118: 1915-1919, 2008; Boengler, Schulz, Heusch, Cardiovasc Res 2009
PI3K
ROS
PKG
mitochondrium
p38
adenosinebradykininopioids, UCN
gp130 TNF-R
JAK
KATP
PKC
mitochondrium
Akt ERK
P70S6K
GPCR PI3K
sGC
Akt
eNOS
NO
eNOS
NO
ANPBNP
GSK3β
aldose reductase
nucleusmitochondrium
STAT3 NO
pGC
MPTP
onlyadenosine H11K
iNOS
MPTP
MnSODSIRT1
CB-R
AMPK
: age-dependent: species-dependent
NPR
Preconditioning: Signal transduction
Cx43
Boengler et al., Cardiovasc Res 67: 234-244, 2005
Boengler et al., Basic Res Cardiol 104: 141-147, 2009
Rodriguez-Sinovas et al., Circ Res 99:93-101, 2006
Heinzel et al., Circ Res 97:583-586, 2005
Görbe et al., Am J Physiol Heart Circ Physiol. 2011 Mar 11
2,5µm
Cytochrome C Connexin 43 Overlay
Mitochondria from human LV tissue
0%
10%
20%
30%
40%
50%
60%
MTR fluorescence [% above control]
WT0
10
20
30
40
50
60
70
80
0
10
20
30
40
50
60
70
80
*p<0.05
Infarct size (% area at risk)
Heinzel et al., Circ Res 97:583-586, 2005
Knockout mice
WT Cx43+/−
PLA Diaz DiazPLACx43 +/−
DiazoxideCx43 +/−
Menadione
*
Mena
Connexin 43 (Cx43), ROS and ischemic preconditioning
Coronary occlusion
Control
Precon(Ischemia/Drugs)
Postcon
60 min
3h5 10 min
3h
3h
Infarct size(% area at risk)
control Precon Postcon
30
25
10
20
15
5
0
P<0.05
Ischemic pre- and post-conditioning
Zhao et al., Am J Physiol 285: H579-H588, 2004
Dog
ROS scavenging and ischemic postconditioning
Rat
Kin et al., Shock 2008
Detection of superoxide anions
contribute to cellular protection
MorphologyFunction
contribute to cellular damage
MorphologyFunction
(Ischemia/Reperfusion Injury)(Postischemic Reperfusion,Heart Failure)
ROS during IRI and protection from it
Minners et al., Cardiovasc Res 47: 68-73, 2000Yue et al., Am J Physiol 281: H590-H595, 2001
Rat heart in vitro
Mitochondrial uncoupling,ROS and IRI
P<0.05
0
5
10
15
20
25
30
35
Placebo IP10 Ascorbic acid IP10 + Ascorbic acid
Infa
rct s
ize
[% a
rea
at ri
sk]
ROS scavenging and ischemic preconditioning
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